-DOCSTART-	O

Number	O
of	O
glucocorticoid	B-protein
receptors	I-protein
in	O
lymphocytes	B-cell_type
and	O
their	O
sensitivity	O
to	O
hormone	O
action	O
.	O

The	O
study	O
demonstrated	O
a	O
decreased	O
level	O
of	O
glucocorticoid	B-protein
receptors	I-protein
(	O
GR	B-protein
)	O
in	O
peripheral	B-cell_type
blood	I-cell_type
lymphocytes	I-cell_type
from	O
hypercholesterolemic	O
subjects	O
,	O
and	O
an	O
elevated	O
level	O
in	O
patients	O
with	O
acute	O
myocardial	O
infarction	O
.	O

In	O
the	O
lymphocytes	B-cell_type
with	O
a	O
high	O
GR	B-protein
number	O
,	O
dexamethasone	O
inhibited	O
[	O
3H	O
]	O
-thymidine	O
and	O
[	O
3H	O
]	O
-acetate	O
incorporation	O
into	O
DNA	O
and	O
cholesterol	O
,	O
respectively	O
,	O
in	O
the	O
same	O
manner	O
as	O
in	O
the	O
control	B-cell_type
cells	I-cell_type
.	O

On	O
the	O
other	O
hand	O
,	O
a	O
decreased	O
GR	B-protein
number	O
resulted	O
in	O
a	O
less	O
efficient	O
dexamethasone	O
inhibition	O
of	O
the	O
incorporation	O
of	O
labeled	O
compounds	O
.	O

These	O
data	O
showed	O
that	O
the	O
sensitivity	O
of	O
lymphocytes	B-cell_type
to	O
glucocorticoids	O
changed	O
only	O
with	O
a	O
decrease	O
of	O
GR	B-protein
level	O
.	O

-DOCSTART-	O

[	O
1	B-protein
,	I-protein
25-Dihydroxyvitamin	I-protein
D3	I-protein
receptors	I-protein
in	O
lymphocytes	B-cell_type
and	O
T-	B-cell_type
and	I-cell_type
B-lymphocyte	I-cell_type
count	O
in	O
patients	O
with	O
glomerulonephritis	O
]	O

Content	O
of	O
receptors	O
to	O
hormonal	O
form	O
of	O
vitamin	O
D3	O
,	O
1.25	O
(	O
OH	O
)	O
2D3	O
,	O
constituted	O
27.3	O
fmole/mg	O
of	O
protein	O
in	O
lymphocytes	B-cell_type
of	O
peripheric	O
blood	O
of	O
children	O
with	O
glomerulonephritis	O
.	O

In	O
the	O
patients	O
concentration	O
of	O
total	O
and	O
ionized	O
form	O
of	O
Ca2+	O
was	O
decreased	O
down	O
to	O
2.04	O
mmole/L	O
and	O
1.09	O
mmole/L	O
,	O
respectively	O
,	O
while	O
an	O
increase	O
in	O
parathormone	O
(	O
PTH	O
)	O
by	O
36	O
%	O
and	O
a	O
distinct	O
decrease	O
in	O
25	O
(	O
OH	O
)	O
D	O
concentration	O
(	O
lower	O
than	O
1.25	O
ng/ml	O
)	O
was	O
found	O
in	O
blood	O
;	O
content	O
of	O
cAMP	O
was	O
also	O
decreased	O
in	O
lymphocytes	O
by	O
33	O
%	O
.	O

At	O
the	O
same	O
time	O
,	O
total	O
content	O
of	O
T	B-cell_type
lymphocytes	I-cell_type
was	O
decreased	O
1.5-fold	O
in	O
peripheric	O
blood	O
.	O

Treatment	O
with	O
I-hydroxyvitamin	O
D3	O
(	O
1-1.5	O
mg	O
daily	O
,	O
within	O
4	O
weeks	O
)	O
led	O
to	O
normalization	O
of	O
total	O
and	O
ionized	O
form	O
of	O
Ca2+	O
and	O
of	O
25	O
(	O
OH	O
)	O
D	O
,	O
but	O
did	O
not	O
affect	O
the	O
PTH	O
content	O
in	O
blood	O
.	O

Concentration	O
of	O
the	O
receptors	B-protein
to	O
1.25	O
(	O
OH	O
)	O
2D3	O
was	O
elevated	O
up	O
to	O
39.7	O
fmole/mg	O
after	O
I	O
week	O
of	O
the	O
treatment	O
,	O
whereas	O
it	O
was	O
decreased	O
to	O
the	O
initial	O
level	O
24.8	O
fmole/mg	O
within	O
4	O
weeks	O
;	O
simultaneous	O
alteration	O
in	O
the	O
cAMP	O
content	O
was	O
observed	O
in	O
lymphocytes	B-cell_type
.	O

Treatment	O
with	O
1-	O
(	O
OH	O
)	O
D3	O
normalized	O
also	O
the	O
T	B-cell_type
lymphocytes	I-cell_type
content	O
in	O
peripheric	O
blood	O
.	O

The	O
data	O
obtained	O
suggest	O
that	O
under	O
conditions	O
of	O
glomerulonephritis	O
only	O
high	O
content	O
of	O
receptors	B-protein
to	O
1.25	O
(	O
OH	O
)	O
2D3	O
in	O
lymphocytes	B-cell_type
enabled	O
to	O
perform	O
the	O
cell	O
response	O
to	O
the	O
hormone	O
effect	O
.	O

-DOCSTART-	O

Tumor	O
and	O
serum	O
beta-2-microglobulin	O
expression	O
in	O
women	O
with	O
breast	O
cancer	O
.	O

To	O
investigate	O
whether	O
the	O
tumor	B-cell_type
expression	O
of	O
beta-2-microglobulin	B-protein
(	O
beta	B-protein
2-M	I-protein
)	O
could	O
serve	O
as	O
a	O
marker	O
of	O
tumor	O
biologic	O
behavior	O
,	O
the	O
authors	O
studied	O
specimens	O
of	O
breast	B-cell_type
carcinomas	I-cell_type
from	O
60	O
consecutive	O
female	O
patients	O
.	O

Presence	O
of	O
beta	B-protein
2-M	I-protein
was	O
analyzed	O
by	O
immunohistochemistry	O
.	O

No	O
significant	O
correlations	O
were	O
found	O
between	O
tumor	B-protein
beta	I-protein
2-M	I-protein
expression	O
and	O
several	O
histologic	O
attributes	O
such	O
as	O
type	O
,	O
histologic	O
and	O
nuclear	O
grades	O
,	O
mitotic	O
index	O
,	O
necrosis	O
,	O
vascular	O
invasion	O
,	O
and	O
lymphocytic	O
infiltration	O
.	O

Likewise	O
,	O
beta	B-protein
2-M	I-protein
was	O
not	O
associated	O
with	O
markers	O
of	O
disease	O
extension	O
such	O
as	O
TNM	O
,	O
(	O
UICC	O
,	O
classification	O
of	O
malignant	B-cell_type
tumors	I-cell_type
)	O
staging	O
and	O
axillary	O
lymph	O
node	O
involvement	O
or	O
with	O
estrogen	O
,	O
progesterone	O
,	O
and	O
glucocorticoid	O
receptor	O
levels	O
.	O

However	O
,	O
there	O
was	O
a	O
significantly	O
positive	O
association	O
between	O
tumor	B-protein
beta	I-protein
2-M	I-protein
expression	O
and	O
the	O
degree	O
of	O
lymphocytic	O
infiltration	O
in	O
the	O
tumor	O
tissue	O
.	O

Beta	B-protein
2-M	I-protein
serum	O
levels	O
were	O
determined	O
by	O
an	O
enzyme-linked	O
immunosorbent	O
assay	O
in	O
samples	O
from	O
22	O
of	O
the	O
above	O
women	O
.	O

Although	O
some	O
of	O
the	O
highest	O
values	O
had	O
been	O
obtained	O
in	O
women	O
with	O
larger	O
(	O
T4	O
)	O
primary	O
tumors	O
,	O
the	O
authors	O
failed	O
to	O
detect	O
any	O
statistical	O
relationship	O
between	O
beta	B-protein
2-M	I-protein
expression	O
in	O
the	O
tumor	O
with	O
serum	O
levels	O
or	O
between	O
serum	B-protein
beta	I-protein
2-M	I-protein
and	O
the	O
above	O
histologic	O
,	O
laboratory	O
,	O
and	O
clinical	O
factors	O
.	O

-DOCSTART-	O

[	O
Preliminary	O
observation	O
of	O
level	O
free-form	B-protein
E	I-protein
receptor	I-protein
levels	O
in	O
serum	O
of	O
normal	O
childbearing-aged	O
and	O
pregnant	O
women	O
]	O

In	O
137	O
cases	O
of	O
childbearing-aged	O
and	O
pregnant	O
women	O
,	O
free	B-protein
form	I-protein
E	I-protein
receptor	I-protein
levels	O
(	O
sE	B-protein
)	O
in	O
serum	O
were	O
measured	O
by	O
ELISA	O
.	O

The	O
level	O
of	O
sE	B-protein
was	O
significantly	O
decreased	O
during	O
the	O
first	O
trimester	O
,	O
slightly	O
higher	O
in	O
the	O
second	O
trimester	O
,	O
and	O
recovered	O
to	O
normal	O
in	O
the	O
third	O
trimester	O
.	O

The	O
level	O
remained	O
lower	O
in	O
29	O
PIH	O
women	O
but	O
appeared	O
higher	O
in	O
overdue	O
pregnancies	O
as	O
compared	O
with	O
the	O
normal	O
3rd	O
trimester	O
range	O
.	O

The	O
results	O
indicate	O
that	O
there	O
is	O
a	O
relationship	O
between	O
a	O
change	O
in	O
T	B-cell_type
cell	I-cell_type
function	O
and	O
pregnancy	O
.	O

-DOCSTART-	O

Kappa	B-protein
B-specific	I-protein
DNA	I-protein
binding	I-protein
proteins	I-protein
:	O
role	O
in	O
the	O
regulation	O
of	O
human	B-DNA
interleukin-2	I-DNA
gene	I-DNA
expression	O
.	O

Transcriptional	O
activation	O
of	O
the	O
human	B-DNA
interleukin-2	I-DNA
(	I-DNA
IL-2	I-DNA
)	I-DNA
gene	I-DNA
,	O
like	O
induction	O
of	O
the	O
IL-2	B-DNA
receptor	I-DNA
alpha	I-DNA
(	I-DNA
IL-2R	I-DNA
alpha	I-DNA
)	I-DNA
gene	I-DNA
and	O
the	O
type	O
1	O
human	O
immunodeficiency	O
virus	O
(	O
HIV-1	O
)	O
,	O
is	O
shown	O
to	O
be	O
modulated	O
by	O
a	O
kappa	B-protein
B-like	I-protein
enhancer	I-protein
element	I-protein
.	O

Mutation	O
of	O
a	O
kappa	B-DNA
B	I-DNA
core	I-DNA
sequence	I-DNA
identified	O
in	O
the	O
IL-2	B-DNA
promoter	I-DNA
(	O
-206	O
to	O
-195	O
)	O
partially	O
inhibits	O
both	O
mitogen-	O
and	O
HTLV-I	O
Tax-mediated	O
activation	O
of	O
this	O
transcription	B-protein
unit	I-protein
and	O
blocks	O
the	O
specific	O
binding	O
of	O
two	O
inducible	B-protein
cellular	I-protein
factors	I-protein
.	O

These	O
kappa	B-protein
B-specific	I-protein
proteins	I-protein
(	O
80	O
to	O
90	O
and	O
50	O
to	O
55	O
kilodaltons	O
)	O
similarly	O
interact	O
with	O
the	O
functional	O
kappa	B-protein
B	I-protein
enhancer	I-protein
present	O
in	O
the	O
IL-2R	B-DNA
alpha	I-DNA
promoter	I-DNA
.	O

These	O
data	O
suggest	O
that	O
these	O
kappa	B-protein
B-specific	I-protein
proteins	I-protein
have	O
a	O
role	O
in	O
the	O
coordinate	O
regulation	O
of	O
this	O
growth	O
factor-growth	O
factor	O
receptor	O
gene	O
system	O
that	O
controls	O
T	B-cell_type
cell	I-cell_type
proliferation	O
.	O

-DOCSTART-	O

Novel	O
region	O
within	O
the	O
V	B-DNA
kappa	I-DNA
gene	I-DNA
promoter	I-DNA
is	O
responsible	O
for	O
tissue	O
and	O
stage-specific	O
expression	O
of	O
immunoglobulin	B-DNA
genes	I-DNA
in	O
human	B-cell_type
lymphoid	I-cell_type
neoplasms	I-cell_type
.	O

Immunoglobulin	B-protein
gene-specific	I-protein
transacting	I-protein
factors	I-protein
have	O
been	O
shown	O
to	O
play	O
a	O
role	O
in	O
lymphoid	O
tissue-specific	O
expression	O
of	O
immunoglobulin	B-DNA
genes	I-DNA
.	O

The	O
role	O
of	O
these	O
factors	O
in	O
B-cell	B-cell_type
differentiation	O
and	O
stage-specific	O
expression	O
of	O
these	O
genes	O
is	O
,	O
however	O
,	O
not	O
fully	O
understood	O
.	O

We	O
have	O
used	O
a	O
model	O
of	O
human	O
lymphoid	O
neoplasia	O
to	O
address	O
this	O
question	O
.	O

Different	O
fragments	O
of	O
unrearranged	B-DNA
human	I-DNA
variable	I-DNA
region	I-DNA
of	O
immunoglobulin	B-DNA
kappa	I-DNA
gene	I-DNA
(	O
V	B-DNA
kappa	I-DNA
)	O
were	O
used	O
for	O
cell-free	O
in	O
vitro	O
transcription	O
and	O
DNA	O
mobility	O
shift	O
assays	O
.	O

Previously	O
described	O
enhancement	O
of	O
in	O
vitro	O
transcription	O
that	O
was	O
only	O
seen	O
with	O
nuclear	O
extracts	O
derived	O
from	O
B-cell	B-cell_type
neoplasms	I-cell_type
corresponding	O
to	O
the	O
late	O
stages	O
of	O
B-cell	B-cell_type
differentiation	O
was	O
shown	O
to	O
be	O
dependent	O
on	O
the	O
actions	O
of	O
these	O
factor	O
(	O
s	O
)	O
on	O
the	O
DNA	O
region	O
within	O
the	O
V	B-DNA
kappa	I-DNA
gene	I-DNA
promoter	I-DNA
.	O

This	O
region	O
is	O
located	O
within	O
the	O
920	O
bp	O
fragment	O
located	O
210	O
bp	O
upstream	O
from	O
the	O
coding	O
region	O
and	O
this	O
fragment	O
represents	O
a	O
possible	O
novel	O
DNA	O
region	O
,	O
which	O
plays	O
a	O
role	O
in	O
the	O
stage-	O
and	O
tissue-specific	O
expression	O
of	O
immunoglobulin	B-DNA
genes	I-DNA
.	O

-DOCSTART-	O

[	O
Determination	O
of	O
the	O
sensitivity	O
to	O
glucocorticoids	B-protein
in	O
vitro	O
]	O

A	O
modified	O
method	O
for	O
the	O
determination	O
of	O
glucocorticoid	B-protein
receptors	I-protein
in	O
human	B-cell_type
lymphocytes	I-cell_type
is	O
suggested	O
.	O

The	O
principal	O
distinction	O
of	O
the	O
method	O
is	O
standardization	O
by	O
the	O
lymphocyte	B-cell_type
count	O
in	O
a	O
sample	O
(	O
1	O
mln	O
)	O
and	O
the	O
labeled	O
hormone	B-protein
concentration	O
.	O

The	O
modification	O
saves	O
time	O
and	O
money	O
,	O
limits	O
the	O
range	O
of	O
the	O
data	O
variations	O
,	O
and	O
makes	O
use	O
of	O
a	O
lesser	O
volume	O
of	O
blood	O
.	O

Examinations	O
of	O
70	O
children	O
aged	O
4	O
to	O
15	O
suffering	O
from	O
the	O
nephrotic	O
form	O
of	O
glomerulonephritis	O
have	O
made	O
it	O
possible	O
to	O
distinguish	O
two	O
groups	O
of	O
patients	O
:	O
with	O
relatively	O
high	O
values	O
of	O
specific	O
binding	O
X	O
=	O
6820.1	O
+/-	O
530.0	O
(	O
n	O
=	O
30	O
,	O
p	O
=	O
0.95	O
,	O
t	O
=	O
2.04	O
)	O
,	O
this	O
corresponding	O
to	O
a	O
clinical	O
form	O
of	O
hormone	B-protein
-sensitive	O
glomerulonephritis	O
,	O
and	O
with	O
relatively	O
low	O
values	O
of	O
specific	O
binding	O
X	O
=	O
1815.2	O
+/-	O
302.8	O
(	O
n	O
=	O
40	O
,	O
p	O
=	O
0.95	O
,	O
t	O
=	O
1.96	O
)	O
,	O
that	O
corresponds	O
to	O
hormone	B-protein
-resistant	O
glomerulonephritis	O
.	O

Dynamic	O
studies	O
have	O
not	O
shown	O
any	O
statistically	O
significant	O
changes	O
in	O
the	O
specific	O
binding	O
values	O
.	O

These	O
results	O
permit	O
regarding	O
the	O
specific	O
binding	O
value	O
as	O
a	O
prognostic	O
criterion	O
in	O
the	O
assessment	O
of	O
corticosteroid	B-protein
therapy	O
;	O
this	O
allows	O
a	O
wide	O
employment	O
of	O
the	O
described	O
method	O
in	O
practical	O
nephrology	O
.	O

-DOCSTART-	O

Octamer-binding	B-protein
proteins	I-protein
from	O
B	B-cell_line
or	I-cell_line
HeLa	I-cell_line
cells	I-cell_line
stimulate	O
transcription	O
of	O
the	O
immunoglobulin	B-DNA
heavy-chain	I-DNA
promoter	I-DNA
in	O
vitro	O
.	O

The	O
B-cell	B-cell_type
-type	O
specificity	O
of	O
the	O
immunoglobulin	B-protein
(	O
Ig	B-protein
)	O
heavy-chain	O
and	O
light-chain	O
promoters	O
is	O
mediated	O
by	O
an	O
octanucleotide	B-DNA
(	I-DNA
OCTA	I-DNA
)	I-DNA
element	I-DNA
,	O
ATGCAAAT	O
,	O
that	O
is	O
also	O
a	O
functional	O
component	O
of	O
other	O
RNA	B-DNA
polymerase	I-DNA
II	I-DNA
promoters	I-DNA
,	O
such	O
as	O
snRNA	O
and	O
histone	O
H2B	O
promoters	O
.	O

Two	O
nuclear	B-protein
proteins	I-protein
that	O
bind	O
specifically	O
and	O
with	O
high	O
affinity	O
to	O
the	O
OCTA	B-DNA
element	I-DNA
have	O
been	O
identified	O
.	O

NF-A1	B-protein
is	O
present	O
in	O
a	O
variety	O
of	O
cell	O
types	O
,	O
whereas	O
the	O
presence	O
of	O
NF-A2	B-protein
is	O
essentially	O
confined	O
to	O
B	B-cell_type
cells	I-cell_type
,	O
leading	O
to	O
the	O
hypothesis	O
that	O
NF-A2	B-protein
activates	O
cell-type-specific	O
transcription	O
of	O
the	O
Ig	B-DNA
promoter	I-DNA
and	O
NF-A1	B-protein
mediates	O
the	O
other	O
responses	O
of	O
the	O
OCTA	B-DNA
element	I-DNA
.	O

Extracts	O
of	O
the	O
B-cell	B-cell_line
line	I-cell_line
,	O
BJA-B	B-cell_line
,	O
contain	O
high	O
levels	O
of	O
NF-A2	B-protein
and	O
specifically	O
transcribe	O
Ig	B-DNA
promoters	I-DNA
.	O

In	O
contrast	O
,	O
extracts	O
from	O
HeLa	B-cell_line
cells	I-cell_line
transcribed	O
the	O
Ig	B-DNA
promoter	I-DNA
poorly	O
.	O

Surprisingly	O
,	O
addition	O
of	O
either	O
affinity-enriched	O
NF-A2	O
or	O
NF-A1	O
to	O
either	O
a	O
HeLa	O
extract	O
or	O
a	O
partially	O
purified	O
reaction	O
system	O
specifically	O
stimulates	O
the	O
Ig	B-DNA
promoter	I-DNA
.	O

This	O
suggests	O
that	O
the	O
constitutive	O
OCTA-binding	B-protein
factor	I-protein
NF-A1	I-protein
can	O
activate	O
transcription	O
of	O
the	O
Ig	B-DNA
promoter	I-DNA
and	O
that	O
B-cell	B-cell_type
-specific	O
transcription	O
of	O
this	O
promoter	O
,	O
at	O
least	O
in	O
vitro	O
,	O
is	O
partially	O
due	O
to	O
a	O
quantitative	O
difference	O
in	O
the	O
amount	O
of	O
OCTA-binding	B-protein
protein	I-protein
.	O

Because	O
NF-A1	B-protein
can	O
stimulate	O
Ig	B-protein
transcription	O
,	O
the	O
inability	O
of	O
this	O
factor	O
to	O
activate	O
in	O
vivo	O
the	O
Ig	B-DNA
promoter	I-DNA
to	O
the	O
same	O
degree	O
as	O
the	O
snRNA	B-DNA
promoters	I-DNA
probably	O
reflects	O
a	O
difference	O
in	O
the	O
context	O
of	O
the	O
OCTA	B-DNA
element	I-DNA
in	O
these	O
two	O
types	O
of	O
promoters	B-DNA
.	O

-DOCSTART-	O

Identification	O
of	O
a	O
putative	O
regulator	O
of	O
early	B-DNA
T	I-DNA
cell	I-DNA
activation	I-DNA
genes	I-DNA
.	O

Molecules	O
involved	O
in	O
the	O
antigen	O
receptor-dependent	O
regulation	O
of	O
early	B-DNA
T	I-DNA
cell	I-DNA
activation	I-DNA
genes	I-DNA
were	O
investigated	O
with	O
the	O
use	O
of	O
functional	O
sequences	O
of	O
the	O
T	B-protein
cell	I-protein
activation-specific	I-protein
enhancer	I-protein
of	O
interleukin-2	B-protein
(	O
IL-2	B-protein
)	O
.	O

One	O
of	O
these	O
sequences	O
forms	O
a	O
protein	O
complex	O
,	O
NFAT-1	B-protein
,	O
specifically	O
with	O
nuclear	O
extracts	O
of	O
activated	O
T	B-cell_type
cells	I-cell_type
.	O

This	O
complex	O
appeared	O
10	O
to	O
25	O
minutes	O
before	O
the	O
activation	O
of	O
the	O
IL-2	B-DNA
gene	I-DNA
.	O

Studies	O
with	O
inhibitors	O
of	O
protein	O
synthesis	O
indicated	O
that	O
the	O
time	O
of	O
synthesis	O
of	O
the	O
activator	O
of	O
the	O
IL-2	B-DNA
gene	I-DNA
in	O
Jurkat	B-cell_line
T	I-cell_line
cells	I-cell_line
corresponds	O
to	O
the	O
time	O
of	O
appearance	O
of	O
NFAT-1	B-protein
.	O

NFAT-1	B-protein
,	O
or	O
a	O
very	O
similar	O
protein	O
,	O
bound	O
functional	O
sequences	O
of	O
the	O
long	B-DNA
terminal	I-DNA
repeat	I-DNA
(	O
LTR	B-DNA
)	O
of	O
the	O
human	O
immunodeficiency	O
virus	O
type	O
1	O
;	O
the	O
LTR	B-DNA
of	O
this	O
virus	O
is	O
known	O
to	O
be	O
stimulated	O
during	O
early	O
T	B-cell_type
cell	I-cell_type
activation	O
.	O

The	O
binding	B-DNA
site	I-DNA
for	O
this	O
complex	O
activated	O
a	O
linked	O
promoter	O
after	O
transfection	O
into	O
antigen	B-cell_type
receptor-activated	I-cell_type
T	I-cell_type
cells	I-cell_type
but	O
not	O
other	O
cell	O
types	O
.	O

These	O
characteristics	O
suggest	O
that	O
NFAT-1	B-protein
transmits	O
signals	O
initiated	O
at	O
the	O
T	B-protein
cell	I-protein
antigen	I-protein
receptor	I-protein
.	O

-DOCSTART-	O

Characterization	O
of	O
thyroid	B-protein
hormone	I-protein
receptors	I-protein
in	O
human	B-cell_line
IM-9	I-cell_line
lymphocytes	I-cell_line
.	O

Although	O
putatively	O
identified	O
more	O
than	O
10	O
years	O
ago	O
,	O
thyroid	B-protein
hormone	I-protein
receptors	I-protein
in	O
human	O
tissues	O
remain	O
poorly	O
characterized	O
.	O

As	O
a	O
first	O
step	O
towards	O
understanding	O
the	O
mechanism	O
of	O
thyroid	O
hormone	O
action	O
in	O
man	O
we	O
have	O
characterized	O
T3	B-protein
binding	I-protein
sites	I-protein
in	O
nuclei	O
of	O
the	O
human	B-cell_line
lymphoblastoid	I-cell_line
line	I-cell_line
,	O
IM-9	B-cell_line
cells	I-cell_line
.	O

In	O
whole	O
cell	O
experiments	O
at	O
37	O
degrees	O
C	O
,	O
nuclear	O
binding	O
of	O
[	B-protein
125I	I-protein
]	I-protein
T3	I-protein
was	O
saturable	O
(	O
Kd	O
34	O
+/-	O
6	O
pmol/l	O
)	O
and	O
of	O
finite	O
capacity	O
(	O
approximately	O
equal	O
to	O
350	O
sites/cell	O
)	O
.	O

The	O
binding	B-protein
sites	I-protein
were	O
extracted	O
from	O
a	O
nuclear	O
pellet	O
by	O
treatment	O
with	O
0.4	O
mol/l	O
KCl	O
and	O
sonication	O
.	O

Separation	O
of	O
bound	O
from	O
free	O
[	B-protein
125I	I-protein
]	I-protein
T3	I-protein
in	O
the	O
extracts	O
was	O
achieved	O
using	O
the	O
calcium	O
phosphate	O
matrix	O
,	O
hydroxyapatite	O
at	O
a	O
concentration	O
of	O
0.3	O
ml	O
of	O
a	O
150	O
g/l	O
slurry	O
.	O

Rectilinear	O
Scatchard	O
plots	O
were	O
obtained	O
only	O
when	O
the	O
hydroxyapatite	O
was	O
washed	O
with	O
a	O
buffer	O
containing	O
0.5	O
%	O
Triton	O
X-100	O
.	O

Under	O
these	O
conditions	O
T3	B-protein
binding	I-protein
sites	I-protein
in	O
the	O
nuclear	O
extracts	O
were	O
present	O
at	O
a	O
concentration	O
of	O
22.4	O
+/-	O
8.6	O
fmol/mg	O
protein	O
and	O
showed	O
an	O
affinity	O
of	O
(	O
Kd	O
,	O
room	O
temperature	O
)	O
140	O
+/-	O
10	O
pmol/l	O
.	O

The	O
same	O
assay	O
system	O
was	O
used	O
to	O
determine	O
the	O
hierarchy	O
of	O
affinities	O
for	O
a	O
range	O
of	O
natural	O
and	O
synthetic	O
analogues	O
.	O

Calling	O
T3	O
100	O
,	O
the	O
order	O
of	O
potencies	O
observed	O
was	O
:	O
Triac	O
,	O
500	O
;	O
3	O
,	O
5-diiodo-3'-isopropylthyronine	O
,	O
89	O
;	O
T4	O
,	O
32	O
;	O
3	O
,	O
5-dimethyl-3'isopropylthyronine	O
2	O
;	O
3	O
,	O
5-T2	O
,	O
0.7	O
,	O
rT3	O
,	O
0.4	O
;	O
3'5'-T2	O
,	O
less	O
than	O
0.01	O
.	O

These	O
results	O
suggest	O
that	O
the	O
T3	B-protein
binding	I-protein
sites	I-protein
present	O
in	O
human	B-cell_line
IM-9	I-cell_line
lymphocyte	I-cell_line
nuclei	O
and	O
extracts	O
thereof	O
are	O
thyroid	B-protein
hormone	I-protein
receptors	I-protein
.	O

These	O
cells	O
may	O
be	O
a	O
useful	O
tool	O
to	O
increase	O
our	O
understanding	O
of	O
human	B-protein
T3	I-protein
receptors	I-protein

-DOCSTART-	O

Definition	O
of	O
T-cell	B-protein
specific	I-protein
DNA-binding	I-protein
factors	I-protein
that	O
interact	O
with	O
a	O
3'-silencer	B-DNA
in	O
the	O
CD4+	B-DNA
T-cell	I-DNA
gene	I-DNA
Rpt-1	I-DNA
.	O

Analysis	O
of	O
the	O
region	O
3	O
'	O
to	O
the	O
CD4+	B-DNA
T-cell	I-DNA
gene	I-DNA
Rpt-1	I-DNA
(	O
encoding	O
regulatory	O
protein	O
T-lymphocyte	B-protein
1	I-protein
)	O
led	O
to	O
the	O
definition	O
of	O
a	O
silencer	B-DNA
element	I-DNA
that	O
inhibits	O
heterologous	B-DNA
gene	I-DNA
expression	O
in	O
certain	O
CD4+	B-cell_type
T-cell	I-cell_type
lines	I-cell_type
but	O
not	O
in	O
B-cell	B-cell_type
or	O
non-lymphoid	B-cell_type
cell	I-cell_type
lines	I-cell_type
.	O

Functional	O
silencer	B-DNA
activity	O
in	O
vivo	O
was	O
associated	O
with	O
the	O
presence	O
of	O
a	O
specific	O
silencer-DNA-protein	B-protein
complex	I-protein
in	O
electrophoretic	O
mobility	O
shift	O
assays	O
with	O
T-cell	B-cell_type
extracts	O
.	O

Formation	O
of	O
this	O
complex	O
was	O
selectively	O
inhibited	O
by	O
the	O
region	O
in	O
HIV-1	B-DNA
containing	O
a	O
silencer	B-DNA
element	I-DNA
.	O

We	O
discuss	O
the	O
possibility	O
that	O
DNA-binding	B-protein
factors	I-protein
may	O
coregulate	O
HIV-1	O
and	O
Rpt-1	O
gene	O
expression	O
through	O
a	O
common	O
transcriptional	B-DNA
silencer	I-DNA
element	I-DNA
.	O

-DOCSTART-	O

Congenital	O
immunodeficiencies	O
associated	O
with	O
absence	O
of	O
HLA	B-protein
class	I-protein
II	I-protein
antigens	I-protein
on	O
lymphocytes	B-cell_type
result	O
from	O
distinct	O
mutations	O
in	O
trans-acting	B-protein
factors	I-protein
.	O

Coordinate	O
regulation	O
of	O
HLA	B-DNA
class	I-DNA
II	I-DNA
gene	I-DNA
expression	O
during	O
development	O
and	O
coinduction	O
of	O
class	B-DNA
II	I-DNA
genes	I-DNA
by	O
soluble	B-protein
factors	I-protein
suggests	O
that	O
common	O
trans-acting	B-protein
factor	I-protein
(	O
s	O
)	O
control	O
expression	O
of	O
these	O
genes	O
.	O

In	O
B-lymphoblastoid	B-cell_type
cell	I-cell_type
lines	I-cell_type
derived	O
from	O
two	O
independent	O
class	O
II-deficient	O
bare	O
lymphocyte	O
syndrome	O
patients	O
,	O
we	O
observed	O
a	O
drastic	O
decrease	O
in	O
transcription	O
rates	O
of	O
the	O
class	B-DNA
II	I-DNA
genes	I-DNA
.	O

When	O
these	O
cell	O
lines	O
are	O
fused	O
,	O
class	B-DNA
II	I-DNA
genes	I-DNA
are	O
reexpressed	O
,	O
indicating	O
that	O
immunodeficiencies	O
in	O
bare	O
lymphocyte	O
syndrome	O
patients	O
are	O
the	O
result	O
of	O
two	O
distinct	O
mutations	O
.	O

Further	O
studies	O
show	O
that	O
genes	O
governing	O
the	O
expression	O
of	O
class	B-protein
II	I-protein
antigens	I-protein
fall	O
into	O
at	O
least	O
three	O
complementation	O
groups	O
;	O
two	O
of	O
these	O
were	O
previously	O
unidentified	O
in	O
mutant	B-cell_line
cell	I-cell_line
lines	I-cell_line
generated	O
in	O
vitro	O
.	O

In	O
addition	O
,	O
we	O
report	O
the	O
identification	O
of	O
two	O
discrete	O
complexes	O
,	O
NFX1.1	B-protein
and	O
NFX1.2	B-protein
,	O
that	O
bind	O
to	O
the	O
DRA	B-DNA
X	I-DNA
consensus	I-DNA
element	I-DNA
.	O

Though	O
the	O
mutation	O
in	O
at	O
least	O
one	O
mutant	B-cell_line
line	I-cell_line
generated	O
in	O
vitro	O
(	O
RJ2.2.5	B-cell_line
)	O
affects	O
products	O
functioning	O
via	O
interaction	O
with	O
the	O
X	B-DNA
box	I-DNA
,	O
clear	O
alterations	O
in	O
either	O
NFX1.1	B-protein
or	O
NFX1.2	B-protein
are	O
not	O
found	O
in	O
any	O
of	O
the	O
mutant	B-cell_line
cell	I-cell_line
lines	I-cell_line
.	O

-DOCSTART-	O

In	O
vivo	O
responsiveness	O
to	O
glucocorticoid	O
correlated	O
with	O
glucocorticoid	B-protein
receptor	I-protein
content	O
in	O
peripheral	B-cell_type
blood	I-cell_type
leukocytes	I-cell_type
in	O
normal	O
humans	O
.	O

Dexamethasone	O
loading	O
tests	O
(	O
0.1	O
mg	O
dexamethasone/kg	O
,	O
iv	O
)	O
were	O
performed	O
in	O
18	O
normal	O
males	O
to	O
evaluate	O
the	O
individual	O
responsiveness	O
to	O
glucocorticoid	O
.	O

There	O
were	O
inter-individual	O
differences	O
in	O
increase	O
in	O
peripheral	B-cell_type
blood	I-cell_type
polymorphonuclear	I-cell_type
leukocyte	I-cell_type
count	O
,	O
decrease	O
in	O
peripheral	B-cell_type
blood	I-cell_type
lymphocyte	I-cell_type
count	O
,	O
and	O
increase	O
in	O
plasma	O
free	O
fatty	O
acids	O
levels	O
after	O
dexamethasone	O
injection	O
.	O

In	O
addition	O
,	O
there	O
was	O
a	O
significant	O
correlation	O
between	O
the	O
maximum	O
increase	O
in	O
polymorphonuclear	B-cell_type
leukocytes	I-cell_type
and	O
the	O
maximum	O
decrease	O
in	O
lymphocytes	B-cell_type
(	O
r	O
=	O
0.7514	O
,	O
p	O
less	O
than	O
0.0003	O
)	O
.	O

Simultaneous	O
measurements	O
of	O
glucocorticoid	B-protein
receptor	I-protein
content	O
by	O
whole-cell	O
assay	O
revealed	O
that	O
glucocorticoid	B-protein
receptor	I-protein
content	O
in	O
polymorphonuclear	B-cell_type
leukocytes	I-cell_type
linearly	O
correlated	O
with	O
that	O
in	O
the	O
corresponding	O
lymphocytes	B-cell_type
(	O
r	O
=	O
0.9482	O
,	O
p	O
less	O
than	O
0.0001	O
)	O
.	O

There	O
were	O
also	O
significant	O
correlations	O
between	O
the	O
maximum	O
increase	O
in	O
polymorphonuclear	B-cell_type
leukocytes	I-cell_type
and	O
glucocorticoid	B-protein
receptor	I-protein
content	O
in	O
polymorphonuclear	B-cell_type
leukocytes	I-cell_type
(	O
r	O
=	O
0.7239	O
,	O
p	O
less	O
than	O
0.0007	O
)	O
,	O
and	O
between	O
the	O
maximum	O
decrease	O
in	O
lymphocytes	B-cell_type
and	O
glucocorticoid	B-protein
receptor	I-protein
content	O
in	O
lymphocytes	B-cell_type
(	O
r	O
=	O
0.7703	O
,	O
p	O
less	O
than	O
0.0002	O
)	O
.	O

These	O
results	O
suggest	O
that	O
individual	O
differences	O
are	O
preserved	O
both	O
in	O
glucocorticoid	O
responsiveness	O
and	O
in	O
glucocorticoid	B-protein
receptor	I-protein
content	O
in	O
peripheral	B-cell_type
blood	I-cell_type
leukocytes	I-cell_type
in	O
normal	O
humans	O
.	O

-DOCSTART-	O

Estradiol	O
receptors	O
in	O
the	O
cytosol	O
of	O
peripheral	B-cell_type
blood	I-cell_type
mononuclear	I-cell_type
cells	I-cell_type
in	O
hepatitis	O
B	O
virus	O
carriers	O
treated	O
with	O
interferon-alpha	B-protein
.	O

Estradiol	B-protein
receptors	I-protein
in	O
the	O
cytosol	O
of	O
peripheral	B-cell_type
blood	I-cell_type
mononuclear	I-cell_type
cells	I-cell_type
and	O
the	O
effects	O
of	O
interferon-alpha	B-protein
(	O
IFN-alpha	B-protein
)	O
on	O
estradiol	B-protein
receptors	I-protein
were	O
studied	O
in	O
asymptomatic	O
hepatitis	O
B	O
virus	O
(	O
HBV	O
)	O
carriers	O
,	O
patients	O
with	O
chronic	O
hepatitis	O
B	O
and	O
normal	O
controls	O
.	O

The	O
level	O
of	O
estradiol	B-protein
receptors	I-protein
in	O
the	O
cytosol	O
of	O
mononuclear	B-cell_type
cells	I-cell_type
was	O
significantly	O
lower	O
in	O
asymptomatic	O
HBV	O
carriers	O
and	O
patients	O
with	O
chronic	O
hepatitis	O
B	O
,	O
compared	O
to	O
normal	O
controls	O
.	O

This	O
low	O
level	O
of	O
cytosol	O
estradiol	B-protein
receptors	I-protein
in	O
patients	O
with	O
chronic	O
hepatitis	O
B	O
was	O
increased	O
by	O
the	O
administration	O
of	O
IFN-alpha	B-protein
.	O

In	O
addition	O
,	O
when	O
peripheral	B-cell_type
blood	I-cell_type
mononuclear	I-cell_type
cells	I-cell_type
from	O
patients	O
with	O
chronic	O
hepatitis	O
B	O
were	O
incubated	O
with	O
IFN-alpha	B-protein
in	O
vitro	O
,	O
the	O
level	O
of	O
cytosol	O
estradiol	B-protein
receptors	I-protein
also	O
increased	O
by	O
increasing	O
the	O
concentration	O
of	O
IFN-alpha	B-protein
.	O

We	O
previously	O
reported	O
that	O
the	O
response	O
of	O
mononuclear	B-cell_type
cells	I-cell_type
to	O
estrogen	O
is	O
impaired	O
in	O
HBV	O
carriers	O
,	O
and	O
our	O
present	O
results	O
suggested	O
that	O
this	O
may	O
be	O
due	O
to	O
the	O
low	O
level	O
of	O
estradiol	B-protein
receptors	I-protein
in	O
the	O
cytosol	O
of	O
mononuclear	B-cell_type
cells	I-cell_type
.	O

-DOCSTART-	O

Association	O
of	O
increased	O
lytic	O
effector	O
cell	O
function	O
with	O
high	O
estrogen	B-protein
receptor	I-protein
levels	O
in	O
tumor-bearing	O
patients	O
with	O
breast	O
cancer	O
.	O

Tumor-bearing	O
patients	O
with	O
breast	O
cancer	O
were	O
assayed	O
for	O
their	O
natural	B-cell_type
killer	I-cell_type
(	I-cell_type
NK	I-cell_type
)	I-cell_type
cell	I-cell_type
activity	O
and	O
for	O
the	O
function	O
of	O
activated	O
cytotoxic	B-cell_type
T-cells	I-cell_type
,	O
as	O
assessed	O
by	O
lectin-dependent	O
cellular	O
cytotoxicity	O
(	O
LDCC	O
)	O
.	O

Tumor-bearing	O
patients	O
with	O
breast	O
cancer	O
had	O
a	O
significant	O
increase	O
in	O
NK	B-cell_type
activity	O
and	O
in	O
LDCC	O
,	O
as	O
compared	O
with	O
healthy	O
control	O
individuals	O
.	O

Although	O
the	O
enhanced	O
NK	B-cell_type
cell	I-cell_type
activity	O
and	O
LDCC	O
were	O
closely	O
associated	O
with	O
high	O
levels	O
(	O
greater	O
than	O
31	O
fmol/mg	O
)	O
of	O
estrogen	B-protein
receptor	I-protein
(	O
ER	B-protein
)	O
content	O
in	O
the	O
primary	O
tumor	O
,	O
no	O
other	O
clinical	O
or	O
histologic	O
correlation	O
between	O
the	O
increase	O
in	O
either	O
parameter	O
of	O
cytotoxic	B-cell_type
effector	I-cell_type
cell	I-cell_type
function	O
could	O
be	O
found	O
.	O

Thus	O
,	O
ER	B-protein
levels	O
greater	O
than	O
31	O
fmol/mg	O
might	O
be	O
associated	O
with	O
increased	O
cytotoxic	B-cell_type
effector	I-cell_type
cell	I-cell_type
function	O
in	O
tumor-bearing	O
patients	O
with	O
breast	O
cancer	O
.	O

-DOCSTART-	O

Properties	O
of	O
glucocorticoid	B-protein
receptors	I-protein
in	O
Epstein-Barr	B-cell_line
virus-transformed	I-cell_line
lymphocytes	I-cell_line
from	O
patients	O
with	O
familial	O
cortisol	O
resistance	O
.	O

In	O
a	O
previous	O
report	O
of	O
two	O
patients	O
with	O
familial	O
glucocorticoid	O
resistance	O
due	O
to	O
reduced	O
numbers	O
of	O
glucocorticoid	B-protein
receptors	I-protein
(	O
GR	B-protein
)	O
,	O
we	O
have	O
shown	O
decreased	O
numbers	O
of	O
GR	B-protein
in	O
peripheral	B-cell_type
mononuclear	I-cell_type
cells	I-cell_type
and	O
cultured	B-cell_line
fibroblasts	I-cell_line
but	O
normal	O
affinity	O
of	O
GR	B-protein
in	O
both	O
patients	O
.	O

In	O
this	O
study	O
,	O
peripheral	B-cell_type
lymphocytes	I-cell_type
from	O
these	O
patients	O
,	O
one	O
patient	O
's	O
son	O
and	O
daughter	O
,	O
and	O
normal	O
subjects	O
were	O
transformed	O
with	O
Epstein-Barr	O
virus	O
.	O

Reduced	O
numbers	O
and	O
normal	O
affinity	O
of	O
GR	B-protein
were	O
found	O
in	O
the	O
Epstein-Barr	B-cell_line
virus-transformed	I-cell_line
lymphocytes	I-cell_line
from	O
both	O
patients	O
while	O
the	O
son	O
and	O
daughter	O
had	O
normal	O
numbers	O
and	O
affinity	O
of	O
GR	B-protein
.	O

The	O
thermal	O
stability	O
of	O
GR	B-protein
and	O
thermal	O
activation	O
of	O
cytosolic	B-protein
receptors	I-protein
in	O
both	O
patients	O
were	O
found	O
to	O
be	O
normal	O
.	O

Although	O
the	O
percentages	O
of	O
nuclear	O
bound	O
GR	B-protein
were	O
similar	O
in	O
both	O
patients	O
and	O
normal	O
controls	O
,	O
the	O
absolute	O
amounts	O
of	O
nuclear	O
bound	O
GR	B-protein
of	O
the	O
patients	O
were	O
about	O
one-half	O
that	O
of	O
normal	O
controls	O
.	O

These	O
abnormal	O
properties	O
of	O
GR	B-protein
(	O
reduced	O
numbers	O
of	O
GR	B-protein
)	O
were	O
preserved	O
in	O
the	O
transformed	B-cell_line
cells	I-cell_line
from	O
the	O
patients	O
.	O

-DOCSTART-	O

Octamer	O
transcription	O
factors	O
1	O
and	O
2	O
each	O
bind	O
to	O
two	O
different	O
functional	B-DNA
elements	I-DNA
in	O
the	O
immunoglobulin	B-DNA
heavy-chain	I-DNA
promoter	I-DNA
.	O

Immunoglobulin	B-DNA
heavy-chain	I-DNA
genes	I-DNA
contain	O
two	O
conserved	B-DNA
sequence	I-DNA
elements	I-DNA
5	O
'	O
to	O
the	O
site	O
of	O
transcription	O
initiation	O
:	O
the	O
octamer	O
ATGCAAAT	O
and	O
the	O
heptamer	O
CTCATGA	O
.	O

Both	O
of	O
these	O
elements	O
are	O
required	O
for	O
normal	B-DNA
cell-specific	I-DNA
promoter	I-DNA
function	O
.	O

The	O
present	O
study	O
demonstrates	O
that	O
both	O
the	O
ubiquitous	B-protein
and	I-protein
lymphoid-cell-specific	I-protein
octamer	I-protein
transcription	I-protein
factors	I-protein
(	O
OTF-1	B-protein
and	O
OTF-2	B-protein
,	O
respectively	O
)	O
interact	O
specifically	O
with	O
each	O
of	O
the	O
two	O
conserved	B-DNA
sequence	I-DNA
elements	I-DNA
,	O
forming	O
either	O
homo-	B-protein
or	I-protein
heterodimeric	I-protein
complexes	I-protein
.	O

This	O
was	O
surprising	O
,	O
since	O
the	O
heptamer	O
and	O
octamer	O
sequence	O
motifs	O
bear	O
no	O
obvious	O
similarity	O
to	O
each	O
other	O
.	O

Binding	O
of	O
either	O
factor	O
to	O
the	O
octamer	B-DNA
element	I-DNA
occurred	O
independently	O
.	O

However	O
,	O
OTF	B-protein
interaction	O
with	O
the	O
heptamer	B-DNA
sequence	I-DNA
appeared	O
to	O
require	O
the	O
presence	O
of	O
an	O
intact	O
octamer	O
motif	O
and	O
occurred	O
with	O
a	O
spacing	O
of	O
either	O
2	B-DNA
or	I-DNA
14	I-DNA
base	I-DNA
pairs	I-DNA
between	O
the	O
two	O
elements	O
,	O
suggesting	O
coordinate	O
binding	O
resulting	O
from	O
protein-protein	O
interactions	O
.	O

The	O
degeneracy	O
in	O
sequences	O
recognized	O
by	O
the	O
OTFs	B-protein
may	O
be	O
important	O
in	O
widening	O
the	O
range	O
over	O
which	O
gene	O
expression	O
can	O
be	O
modulated	O
and	O
in	O
establishing	O
cell	O
type	O
specificity	O
.	O

-DOCSTART-	O

Identification	O
of	O
a	O
novel	O
lymphoid	B-protein
specific	I-protein
octamer	I-protein
binding	I-protein
protein	I-protein
(	O
OTF-2B	B-protein
)	O
by	O
proteolytic	O
clipping	O
bandshift	O
assay	O
(	O
PCBA	O
)	O
.	O

The	O
octamer	O
sequence	O
ATGCAAAT	O
is	O
found	O
in	O
the	O
promoters	O
of	O
immunoglobulin	B-DNA
(	I-DNA
Ig	I-DNA
)	I-DNA
heavy	I-DNA
and	I-DNA
light	I-DNA
chain	I-DNA
genes	I-DNA
and	O
in	O
the	O
heavy	B-DNA
chain	I-DNA
enhancer	I-DNA
and	O
is	O
a	O
major	O
determinant	O
of	O
the	O
cell	O
type	O
specific	O
expression	O
of	O
Ig	B-DNA
genes	I-DNA
in	O
B	B-cell_type
cells	I-cell_type
.	O

An	O
apparent	O
paradox	O
is	O
that	O
the	O
same	O
sequence	O
serves	O
as	O
an	O
upstream	B-DNA
promoter	I-DNA
or	I-DNA
enhancer	I-DNA
element	I-DNA
in	O
a	O
variety	O
of	O
housekeeping	B-DNA
genes	I-DNA
such	O
as	O
the	O
histone	B-DNA
H2B	I-DNA
and	I-DNA
U	I-DNA
snRNA	I-DNA
genes	I-DNA
.	O

The	O
differential	O
usage	O
of	O
this	O
regulatory	O
sequence	O
motif	O
is	O
thought	O
to	O
be	O
mediated	O
by	O
different	O
species	O
of	O
octamer	B-protein
binding	I-protein
proteins	I-protein
.	O

One	O
species	O
of	O
100	O
kd	O
,	O
designated	O
OTF-1	B-protein
,	O
is	O
present	O
in	O
all	O
cell	O
types	O
and	O
may	O
exert	O
its	O
activating	O
function	O
only	O
when	O
it	O
can	O
interact	O
with	O
additional	O
adjacent	O
transcription	B-protein
factors	I-protein
.	O

The	O
lymphoid	B-protein
cell	I-protein
specific	I-protein
octamer	I-protein
binding	I-protein
protein	I-protein
of	O
60	O
kd	O
(	O
OTF-2A	B-protein
)	O
specifically	O
stimulates	O
Ig	B-DNA
promoters	I-DNA
which	O
consist	O
essentially	O
of	O
a	O
TATA-box	B-DNA
and	O
an	O
octamer	B-DNA
sequence	I-DNA
upstream	O
of	O
it	O
.	O

Here	O
we	O
present	O
evidence	O
for	O
yet	O
another	O
B	B-protein
cell	I-protein
specific	I-protein
octamer	I-protein
binding	I-protein
protein	I-protein
of	O
75	O
kd	O
(	O
OTF-2B	B-protein
)	O
.	O

From	O
several	O
findings	O
,	O
including	O
the	O
absence	O
of	O
OTF-2B	B-protein
(	O
but	O
not	O
OTF-2A	B-protein
)	O
from	O
a	O
lymphocyte	B-cell_line
line	I-cell_line
that	O
can	O
not	O
respond	O
to	O
the	O
IgH	B-DNA
enhancer	I-DNA
,	O
we	O
propose	O
a	O
role	O
of	O
the	O
novel	B-protein
octamer	I-protein
factor	I-protein
in	O
the	O
long	O
range	O
activation	O
by	O
the	O
IgH	B-DNA
enhancer	I-DNA
.	O

We	O
have	O
used	O
the	O
proteolytic	O
clipping	O
bandshift	O
assay	O
(	O
PCBA	O
)	O
technique	O
to	O
distinguish	O
the	O
three	O
different	O
forms	O
found	O
in	O
B	B-cell_type
cells	I-cell_type
.	O

This	O
analysis	O
indicates	O
that	O
the	O
75	O
kd-species	O
OTF-2B	B-protein
is	O
closely	O
related	O
to	O
the	O
60	B-protein
kd	I-protein
species	I-protein
OTF-2A	I-protein
.	O

-DOCSTART-	O

Inhibition	O
of	O
interleukin	B-protein
2	I-protein
-induced	O
proliferation	O
of	O
cloned	B-cell_line
murine	I-cell_line
T	I-cell_line
cells	I-cell_line
by	O
glucocorticoids	O
.	O

Possible	O
involvement	O
of	O
an	O
inhibitory	B-protein
protein	I-protein
.	O

The	O
ability	O
of	O
glucocorticoids	O
to	O
inhibit	O
interleukin	B-protein
2	I-protein
(	O
IL	B-protein
2	I-protein
)	O
-induced	O
T	B-cell_type
cell	I-cell_type
proliferation	O
in	O
two	O
cytotoxic	B-cell_line
T	I-cell_line
cell	I-cell_line
(	I-cell_line
CTL	I-cell_line
)	I-cell_line
clones	I-cell_line
has	O
been	O
studied	O
.	O

A	O
complete	O
inhibition	O
of	O
DNA	O
synthesis	O
by	O
dexamethasone	O
(	O
Dx	O
)	O
could	O
be	O
observed	O
when	O
IL	B-cell_line
2-depleted	I-cell_line
cultures	I-cell_line
of	O
CTL	B-cell_line
were	O
either	O
incubated	O
for	O
6	O
h	O
with	O
the	O
hormone	B-protein
prior	O
to	O
the	O
addition	O
of	O
IL	B-protein
2	I-protein
or	O
treated	O
simultaneously	O
with	O
Dx	O
and	O
a	O
low	O
concentration	O
of	O
IL	B-protein
2	I-protein
.	O

No	O
significant	O
reduction	O
in	O
the	O
number	O
and	O
affinity	O
of	O
IL	B-protein
2	I-protein
receptors	I-protein
was	O
seen	O
after	O
6	O
h	O
incubation	O
with	O
Dx	O
.	O

The	O
order	O
of	O
potency	O
observed	O
with	O
the	O
different	O
steroids	O
indicated	O
that	O
this	O
inhibitory	O
effect	O
was	O
mediated	O
through	O
binding	O
to	O
a	O
specific	O
glucocorticoid	B-protein
receptor	I-protein
.	O

The	O
action	O
of	O
these	O
hormones	B-protein
possibly	O
involves	O
the	O
synthesis	O
of	O
an	O
inhibitory	B-protein
protein	I-protein
(	O
s	O
)	O
,	O
since	O
the	O
presence	O
of	O
cycloheximide	O
during	O
the	O
incubation	O
with	O
Dx	O
prevented	O
the	O
inhibition	O
of	O
DNA	O
synthesis	O
.	O

Furthermore	O
,	O
supernatant	O
from	O
Dx-treated	O
CTL	B-cell_line
contained	O
a	O
nondialyzable	O
factor	O
which	O
inhibited	O
DNA	O
synthesis	O
and	O
cell	O
growth	O
of	O
CTL	B-cell_line
clones	I-cell_line
induced	O
by	O
IL	B-protein
2	I-protein
.	O

Blocking	O
of	O
IL	B-protein
2	I-protein
synthesis	O
and	O
IL	B-protein
2	I-protein
receptor	I-protein
formation	O
have	O
been	O
proposed	O
as	O
one	O
of	O
the	O
major	O
mechanisms	O
of	O
glucocorticoid-induced	O
immunosuppression	O
.	O

Our	O
results	O
indicate	O
that	O
these	O
hormones	B-protein
may	O
also	O
affect	O
T	B-cell_type
cell	I-cell_type
proliferation	O
by	O
inhibiting	O
IL	B-protein
2	I-protein
activity	O
.	O

-DOCSTART-	O

Identification	O
and	O
purification	O
of	O
a	O
human	B-protein
immunoglobulin-enhancer-binding	I-protein
protein	I-protein
(	O
NF-kappa	B-protein
B	I-protein
)	O
that	O
activates	O
transcription	O
from	O
a	O
human	B-DNA
immunodeficiency	I-DNA
virus	I-DNA
type	I-DNA
1	I-DNA
promoter	I-DNA
in	O
vitro	O
.	O

The	O
enhancer-binding	B-protein
factor	I-protein
NF-kappa	I-protein
B	I-protein
,	O
which	O
is	O
found	O
only	O
in	O
cells	O
that	O
transcribe	O
immunoglobulin	B-DNA
light	I-DNA
chain	I-DNA
genes	I-DNA
,	O
has	O
been	O
purified	O
from	O
nuclear	O
extracts	O
of	O
Namalwa	B-cell_line
cells	I-cell_line
(	O
human	B-cell_line
Burkitt	I-cell_line
lymphoma	I-cell_line
cells	I-cell_line
)	O
by	O
sequence-specific	O
DNA	O
affinity	O
chromatography	O
.	O

The	O
purified	O
NF-kappa	B-protein
B	I-protein
has	O
been	O
identified	O
as	O
a	O
51-kDa	B-protein
polypeptide	I-protein
by	O
UV-crosslinking	O
analysis	O
.	O

``	O
Footprint	O
''	O
and	O
methylation-interference	O
analyses	O
have	O
shown	O
that	O
purified	O
NF-kappa	B-protein
B	I-protein
has	O
a	O
binding	O
activity	O
specific	O
for	O
the	O
kappa	B-DNA
light	I-DNA
chain	I-DNA
enhancer	I-DNA
sequence	I-DNA
.	O

The	O
purified	O
factor	O
activated	O
in	O
vitro	O
transcription	O
of	O
the	O
human	B-DNA
immunodeficiency	I-DNA
virus	I-DNA
type	I-DNA
I	I-DNA
promoter	I-DNA
by	O
binding	O
to	O
an	O
upstream	B-DNA
NF-kappa	I-DNA
B-binding	I-DNA
site	I-DNA

-DOCSTART-	O

Lymphocyte	B-cell_type
glucocorticoid	B-protein
receptor	I-protein
binding	O
in	O
depression	O
:	O
normal	O
values	O
following	O
recovery	O
.	O

The	O
number	O
of	O
glucocorticoid	B-protein
receptor	I-protein
sites	O
in	O
lymphocytes	B-cell_type
and	O
plasma	O
cortisol	O
concentrations	O
were	O
measured	O
in	O
20	O
patients	O
who	O
had	O
recovered	O
from	O
major	O
depressive	O
disorder	O
and	O
20	O
healthy	O
control	O
subjects	O
.	O

The	O
number	O
of	O
glucocorticoid	B-protein
receptor	I-protein
sites	O
in	O
lymphocytes	B-cell_type
from	O
the	O
recovered	O
depressed	O
group	O
was	O
not	O
significantly	O
different	O
from	O
that	O
of	O
the	O
control	O
group	O
.	O

Although	O
the	O
mean	O
plasma	O
cortisol	O
concentration	O
in	O
recovered	O
depressives	O
was	O
higher	O
than	O
in	O
control	O
subjects	O
,	O
the	O
difference	O
only	O
just	O
reached	O
significance	O
.	O

This	O
study	O
shows	O
that	O
the	O
reduction	O
in	O
glucocorticoid	B-protein
receptor	I-protein
numbers	O
which	O
occurs	O
during	O
acute	O
depressive	O
illness	O
does	O
not	O
persist	O
on	O
recovery	O
and	O
is	O
,	O
therefore	O
,	O
state-dependent	O
.	O

-DOCSTART-	O

Identification	O
and	O
purification	O
of	O
a	O
human	B-protein
lymphoid-specific	I-protein
octamer-binding	I-protein
protein	I-protein
(	O
OTF-2	B-protein
)	O
that	O
activates	O
transcription	O
of	O
an	O
immunoglobulin	B-DNA
promoter	I-DNA
in	O
vitro	O
.	O

The	O
octamer	O
sequence	O
5'-ATGCAAAT	O
,	O
in	O
either	O
orientation	O
,	O
serves	O
as	O
an	O
upstream	B-DNA
element	I-DNA
in	O
a	O
variety	O
of	O
promoters	B-DNA
and	O
also	O
occurs	O
as	O
a	O
modular	B-DNA
enhancer	I-DNA
element	I-DNA
.	O

It	O
is	O
of	O
particular	O
interest	O
in	O
immunoglobulin	B-DNA
genes	I-DNA
since	O
it	O
is	O
found	O
in	O
the	O
upstream	B-DNA
regions	I-DNA
of	O
all	O
heavy	B-DNA
and	I-DNA
light	I-DNA
chain	I-DNA
promoters	I-DNA
and	O
in	O
the	O
heavy	B-DNA
chain	I-DNA
enhancer	I-DNA
,	O
both	O
of	O
which	O
are	O
known	O
to	O
be	O
necessary	O
for	O
cell-specific	O
expression	O
.	O

We	O
report	O
here	O
the	O
chromatographic	O
separation	O
of	O
ubiquitous	B-protein
and	I-protein
B	I-protein
cell-specific	I-protein
octamer-binding	I-protein
proteins	I-protein
.	O

The	O
B	B-protein
cell	I-protein
factor	I-protein
was	O
purified	O
to	O
homogeneity	O
using	O
affinity	O
chromatography	O
and	O
consists	O
of	O
three	O
peptides	O
of	O
62	O
,	O
61	O
,	O
and	O
58.5	O
+/-	O
1.5	O
kd	O
.	O

Each	O
of	O
the	O
polypeptides	O
was	O
renatured	O
after	O
SDS-PAGE	O
and	O
shown	O
to	O
bind	O
to	O
the	O
octamer	O
sequence	O
.	O

The	O
specific	O
DNA	O
binding	O
activity	O
of	O
the	O
pure	O
B	B-protein
cell-specific	I-protein
factor	I-protein
was	O
indistinguishable	O
from	O
that	O
of	O
the	O
affinity-purified	B-protein
ubiquitous	I-protein
factor	I-protein
.	O

This	O
B	B-protein
cell-specific	I-protein
octamer-binding	I-protein
factor	I-protein
,	O
in	O
pure	O
form	O
,	O
activated	O
transcription	O
from	O
a	O
kappa	B-DNA
light	I-DNA
chain	I-DNA
promoter	I-DNA
in	O
vitro	O
,	O
thus	O
demonstrating	O
that	O
it	O
is	O
indeed	O
a	O
B	B-protein
cell-specific	I-protein
transcription	I-protein
factor	I-protein
for	O
this	O
gene	O
.	O

In	O
addition	O
to	O
the	O
ubiquitous	B-protein
and	I-protein
B	I-protein
cell-specific	I-protein
octamer-binding	I-protein
factors	I-protein
,	O
we	O
identified	O
several	O
additional	O
proteins	O
,	O
one	O
of	O
which	O
is	O
B	B-cell_type
cell	I-cell_type
-specific	O
,	O
that	O
interact	O
with	O
the	O
kappa	B-DNA
promoter	I-DNA
.	O

-DOCSTART-	O

Decreased	O
deoxyribonucleic	O
acid	O
binding	O
of	O
glucocorticoid-receptor	B-protein
complex	I-protein
in	O
cultured	O
skin	B-cell_line
fibroblasts	I-cell_line
from	O
a	O
patient	O
with	O
the	O
glucocorticoid	O
resistance	O
syndrome	O
.	O

A	O
patient	O
with	O
the	O
syndrome	O
of	O
glucocorticoid	O
resistance	O
was	O
studied	O
.	O

A	O
27-yr-old	O
woman	O
initially	O
was	O
diagnosed	O
as	O
having	O
Cushing	O
's	O
disease	O
,	O
based	O
on	O
the	O
findings	O
of	O
high	O
plasma	O
ACTH	O
and	O
serum	O
cortisol	O
levels	O
,	O
increased	O
urinary	O
cortisol	O
secretion	O
,	O
resistance	O
to	O
adrenal	O
suppression	O
with	O
dexamethasone	O
,	O
and	O
bilateral	O
adrenal	O
hyperplasia	O
by	O
computed	O
tomography	O
and	O
scintigraphy	O
of	O
the	O
adrenal	O
glands	O
.	O

However	O
,	O
she	O
had	O
no	O
signs	O
or	O
symptoms	O
of	O
Cushing	O
's	O
syndrome	O
.	O

During	O
a	O
5-yr	O
follow-up	O
,	O
no	O
clinical	O
abnormalities	O
developed	O
,	O
although	O
hypercortisolism	O
persisted	O
.	O

End-organ	O
resistance	O
to	O
cortisol	O
was	O
suspected	O
.	O

To	O
explain	O
the	O
end-organ	O
resistance	O
to	O
cortisol	O
,	O
the	O
glucocorticoid	B-protein
receptors	I-protein
(	O
GR	B-protein
)	O
in	O
peripheral	B-cell_type
mononuclear	I-cell_type
leukocytes	I-cell_type
and	O
cultured	B-cell_line
skin	I-cell_line
fibroblasts	I-cell_line
from	O
a	O
forearm	O
skin	O
biopsy	O
were	O
characterized	O
and	O
compared	O
with	O
the	O
results	O
of	O
similar	O
studies	O
in	O
normal	O
subjects	O
.	O

The	O
patient	O
's	O
GR	B-protein
in	O
whole	O
cell	O
assays	O
had	O
an	O
increased	O
dissociation	O
constant	O
(	O
Kd	O
)	O
.	O

In	O
the	O
cytosol	O
of	O
cultured	B-cell_line
skin	I-cell_line
fibroblasts	I-cell_line
from	O
the	O
patient	O
,	O
there	O
was	O
also	O
decreased	O
binding	O
capacity	O
.	O

The	O
thermal	O
stability	O
and	O
the	O
sedimentation	O
coefficient	O
in	O
a	O
sucrose	O
density	O
gradient	O
of	O
the	O
receptors	O
in	O
the	O
cytosol	O
of	O
cultured	B-cell_line
skin	I-cell_line
fibroblasts	I-cell_line
from	O
the	O
patient	O
and	O
normal	O
subjects	O
were	O
similar	O
.	O

GR	B-cell_line
complex	I-cell_line
activation	I-cell_line
,	O
analyzed	O
by	O
DEAE-cellulose	B-cell_line
chromatography	I-cell_line
,	O
was	O
decreased	O
in	O
the	O
patient	O
.	O

DNA	B-DNA
binding	O
of	O
the	O
GR	B-protein
complex	I-protein
after	O
temperature-induced	O
activation	O
was	O
lower	O
in	O
the	O
patient	O
than	O
in	O
normal	O
subjects	O
.	O

Nuclear	O
translocation	O
of	O
GR	B-protein
complexes	I-protein
from	O
the	O
patient	O
was	O
also	O
slightly	O
decreased	O
.	O

These	O
results	O
suggest	O
that	O
the	O
patient	O
's	O
glucocorticoid	O
resistance	O
was	O
due	O
to	O
a	O
decrease	O
in	O
the	O
affinity	O
of	O
the	O
receptor	O
for	O
glucocorticoids	O
and	O
a	O
decrease	O
in	O
the	O
binding	O
of	O
the	O
GR	B-protein
complex	I-protein
to	O
DNA	B-DNA
.	O

-DOCSTART-	O

Granulocyte-macrophage	B-protein
colony-stimulating	I-protein
factor	I-protein
.	O

Sensitive	O
and	O
receptor-mediated	O
regulation	O
by	O
1	O
,	O
25-dihydroxyvitamin	O
D3	O
in	O
normal	O
human	B-cell_type
peripheral	I-cell_type
blood	I-cell_type
lymphocytes	I-cell_type
.	O

We	O
show	O
that	O
1	O
,	O
25-dihydroxyvitamin	O
D3	O
(	O
1	O
,	O
25	O
[	O
OH	O
]	O
2D3	O
)	O
,	O
the	O
most	O
hormonally	O
active	O
metabolite	O
of	O
vitamin	O
D3	O
,	O
modulates	O
sensitively	O
and	O
specifically	O
both	O
the	O
protein	O
and	O
messenger	B-RNA
RNA	I-RNA
accumulation	O
of	O
the	O
multilineage	B-protein
growth	I-protein
factor	I-protein
granulocyte-macrophage	B-protein
colony-stimulating	I-protein
factor	I-protein
(	O
GM-CSF	B-protein
)	O
.	O

The	O
regulation	O
of	O
GM-CSF	B-protein
expression	O
is	O
seen	O
in	O
both	O
normal	B-cell_type
human	I-cell_type
mitogen-activated	I-cell_type
T	I-cell_type
lymphocytes	I-cell_type
and	O
T	B-cell_line
lymphocytes	I-cell_line
from	I-cell_line
a	I-cell_line
line	I-cell_line
(	I-cell_line
S-LB1	I-cell_line
)	I-cell_line
transformed	I-cell_line
with	I-cell_line
human	I-cell_line
T	I-cell_line
cell	I-cell_line
lymphotropic	I-cell_line
virus	I-cell_line
1	I-cell_line
(	O
HTLV-1	O
)	O
.	O

In	O
contrast	O
,	O
cells	O
from	O
a	O
HTLV-1	B-cell_line
transformed	I-cell_line
T	I-cell_line
lymphocyte	I-cell_line
line	I-cell_line
(	O
Ab-VDR	B-cell_line
)	O
established	O
from	O
a	O
patient	O
with	O
vitamin	O
D-resistant	O
rickets	O
type	O
II	O
with	O
undetectable	O
1	B-protein
,	I-protein
25	I-protein
(	I-protein
OH	I-protein
)	I-protein
2D3	I-protein
cellular	I-protein
receptors	I-protein
are	O
resistant	O
to	O
the	O
action	O
of	O
1	O
,	O
25	O
(	O
OH	O
)	O
2D3	O
.	O

Inhibition	O
of	O
GM-CSF	B-protein
expression	O
by	O
1	O
,	O
25	O
(	O
OH	O
)	O
2D3	O
can	O
occur	O
independently	O
of	O
interleukin	B-protein
2	I-protein
regulation	O
and	O
is	O
probably	O
mediated	O
through	O
cellular	B-protein
1	I-protein
,	I-protein
25	I-protein
(	I-protein
OH	I-protein
)	I-protein
2D3	I-protein
receptors	I-protein
.	O

We	O
conclude	O
that	O
1	O
,	O
25	O
(	O
OH	O
)	O
2D3	O
may	O
be	O
important	O
in	O
the	O
physiology	O
of	O
hematopoiesis	O
.	O

-DOCSTART-	O

Altered	O
interaction	O
between	O
triiodothyronine	O
and	O
its	O
nuclear	B-protein
receptors	I-protein
in	O
absence	O
of	O
cortisol	O
:	O
a	O
proposed	O
mechanism	O
for	O
increased	O
thyrotropin	O
secretion	O
in	O
corticosteroid	O
deficiency	O
states	O
.	O

Thyroid	O
hormones	O
occasionally	O
appear	O
less	O
effective	O
when	O
administered	O
alone	O
to	O
patients	O
with	O
panhypopituitarism	O
,	O
and	O
manifestations	O
suggestive	O
of	O
hypothyroidism	O
have	O
been	O
reported	O
in	O
patients	O
suffering	O
from	O
untreated	O
Addison	O
's	O
disease	O
.	O

In	O
the	O
latter	O
condition	O
,	O
thyrotropin	O
secretion	O
is	O
increased	O
:	O
this	O
occurs	O
already	O
after	O
as	O
little	O
as	O
2	O
days	O
of	O
temporary	O
withdrawal	O
of	O
therapy	O
with	O
substitution	O
doses	O
of	O
corticosteroids	O
while	O
circulating	O
levels	O
of	O
thyroid	O
hormones	O
remain	O
within	O
normal	O
limits	O
.	O

Therefore	O
,	O
a	O
possible	O
role	O
of	O
cortisol	O
in	O
interaction	O
between	O
triiodothyronine	O
and	O
its	O
nuclear	B-protein
receptors	I-protein
was	O
examined	O
at	O
the	O
level	O
of	O
circulating	O
lymphocytes	O
obtained	O
from	O
patients	O
with	O
primary	O
or	O
secondary	O
adrenocortical	O
failure	O
.	O

The	O
affinity	O
of	O
these	O
receptors	O
was	O
found	O
to	O
be	O
decreased	O
,	O
by	O
more	O
than	O
50	O
%	O
on	O
average	O
,	O
in	O
the	O
absence	O
of	O
cortisol	O
treatments	O
.	O

This	O
change	O
was	O
promptly	O
corrected	O
upon	O
resumption	O
of	O
therapy	O
.	O

The	O
number	O
of	O
binding	O
sites	O
was	O
not	O
significantly	O
modified	O
.	O

The	O
influence	O
of	O
cortisol	O
on	O
thyroid	B-protein
hormone	I-protein
receptors	I-protein
discussed	O
here	O
might	O
account	O
for	O
the	O
clinical	O
observations	O
mentioned	O
above	O
.	O

-DOCSTART-	O

Inhibition	O
by	O
cortisol	O
of	O
human	B-cell_type
natural	I-cell_type
killer	I-cell_type
(	I-cell_type
NK	I-cell_type
)	I-cell_type
cell	I-cell_type
activity	O
.	O

The	O
effects	O
of	O
cortisol	O
on	O
the	O
natural	B-cell_type
killer	I-cell_type
(	O
NK	O
)	O
activity	O
of	O
human	B-cell_type
peripheral	I-cell_type
blood	I-cell_type
mononuclear	I-cell_type
(	I-cell_type
PBM	I-cell_type
)	I-cell_type
cells	I-cell_type
were	O
studied	O
in	O
vitro	O
using	O
a	O
direct	O
4-h	O
51Cr-release	O
assay	O
and	O
K	B-cell_line
562	I-cell_line
cell	I-cell_line
line	I-cell_line
as	O
a	O
target	O
.	O

Preincubation	O
for	O
20	O
h	O
of	O
PBM	B-cell_type
cells	I-cell_type
drawn	O
from	O
healthy	O
donors	O
with	O
1	O
X	O
10	O
(	O
-8	O
)	O
to	O
1	O
X	O
10	O
(	O
-5	O
)	O
M	O
cortisol	O
resulted	O
in	O
a	O
significant	O
decrease	O
of	O
NK	B-cell_type
cell	I-cell_type
activity	O
.	O

The	O
magnitude	O
of	O
the	O
suppression	O
was	O
directly	O
related	O
to	O
the	O
steroid	O
concentration	O
and	O
inversely	O
related	O
to	O
the	O
number	O
of	O
effector	B-cell_type
cells	I-cell_type
.	O

Cortisol	O
was	O
able	O
to	O
minimize	O
the	O
enhancement	O
of	O
NK	O
cytotoxicity	O
obtainable	O
in	O
the	O
presence	O
of	O
immune	O
interferon	O
(	O
IFN-gamma	O
)	O
.	O

A	O
significantly	O
higher	O
suppression	O
was	O
achieved	O
after	O
sequential	O
exposure	O
of	O
PBM	B-cell_type
cells	I-cell_type
to	O
cortisol	O
and	O
equimolar	O
levels	O
of	O
prostaglandin	O
E2	O
(	O
PgE2	O
)	O
.	O

The	O
concomitant	O
incubation	O
with	O
theophylline	O
and	O
isobutyl-methylxanthine	O
failed	O
to	O
enhance	O
the	O
cortisol-induced	O
suppression	O
,	O
whereas	O
PgE2-dependent	O
inhibition	O
significantly	O
increased	O
after	O
exposure	O
of	O
PBM	B-cell_type
cells	I-cell_type
to	O
methyl-xanthines	O
.	O

The	O
inhibitory	O
effect	O
of	O
cortisol	O
was	O
partially	O
or	O
totally	O
prevented	O
by	O
the	O
concomitant	O
incubation	O
with	O
equimolar	O
amounts	O
of	O
11-deoxycortisol	O
and	O
RU	O
486	O
but	O
not	O
of	O
progesterone	O
.	O

Treatment	O
of	O
NK	B-cell_type
effectors	I-cell_type
with	O
a	O
monoclonal	B-protein
anti-human	I-protein
corticosteroid-binding	I-protein
globulin	I-protein
(	I-protein
CBG	I-protein
)	I-protein
antibody	I-protein
produced	O
an	O
enhancement	O
of	O
the	O
spontaneous	O
NK	B-cell_type
activity	O
and	O
a	O
partial	O
suppression	O
of	O
cortisol-mediated	O
effects	O
.	O

Our	O
results	O
suggest	O
that	O
endogenous	O
glucocorticoids	O
play	O
a	O
role	O
in	O
the	O
regulation	O
of	O
NK	O
cell-mediated	O
cytotoxicity	O
.	O

Since	O
the	O
effect	O
of	O
cortisol	O
was	O
additive	O
to	O
that	O
of	O
PgE2	O
and	O
was	O
not	O
changed	O
by	O
phosphodiesterase	B-protein
inhibitors	O
,	O
it	O
is	O
conceivable	O
that	O
the	O
hormone	O
acts	O
at	O
a	O
level	O
different	O
from	O
the	O
adenylate	B-protein
cyclase	I-protein
-	O
phosphodiesterase	B-protein
system	O
.	O

Data	O
obtained	O
with	O
the	O
use	O
of	O
antiglucocorticoids	O
and	O
the	O
anti-CBG	B-protein
antibody	I-protein
are	O
compatible	O
with	O
a	O
role	O
both	O
of	O
high-affinity	O
glucocorticoid	B-protein
receptors	I-protein
and	O
of	O
CBG	B-protein
in	O
mediating	O
cortisol	O
action	O
on	O
the	O
human	B-cell_type
NK	I-cell_type
cell	I-cell_type
activity	O
.	O

-DOCSTART-	O

Interaction	O
of	O
cell-type-specific	B-protein
nuclear	I-protein
proteins	I-protein
with	O
immunoglobulin	B-DNA
VH	I-DNA
promoter	I-DNA
region	I-DNA
sequences	I-DNA
.	O

All	O
human	B-DNA
and	I-DNA
murine	I-DNA
immunoglobulin	I-DNA
heavy	I-DNA
chain	I-DNA
variable	I-DNA
region	I-DNA
(	I-DNA
VH	I-DNA
)	I-DNA
genes	I-DNA
contain	O
the	O
sequence	O
ATGCAAAT	O
approximately	O
70	O
nucleotides	O
5	O
'	O
from	O
the	O
site	O
of	O
transcription	O
initiation	O
.	O

This	O
octanucleotide	O
,	O
in	O
reverse	O
orientation	O
,	O
is	O
also	O
found	O
in	O
all	O
light	B-DNA
chain	I-DNA
variable	I-DNA
region	I-DNA
(	I-DNA
VL	I-DNA
)	I-DNA
genes	I-DNA
,	O
and	O
in	O
the	O
immunoglobulin	B-DNA
heavy	I-DNA
chain	I-DNA
transcriptional	I-DNA
enhancer	I-DNA
.	O

Transfection	O
studies	O
have	O
established	O
that	O
this	O
octamer	O
is	O
involved	O
in	O
the	O
lymphoid-specific	O
transcription	O
of	O
immunoglobulin	B-DNA
genes	I-DNA
.	O

Octamer-containing	B-DNA
fragments	I-DNA
have	O
been	O
reported	O
to	O
bind	O
a	O
factor	O
present	O
in	O
nuclear	O
extracts	O
of	O
human	B-cell_line
cell	I-cell_line
lines	I-cell_line
;	O
however	O
,	O
identical	O
binding	O
activity	O
was	O
detected	O
in	O
both	O
B	B-cell_line
lymphoid	I-cell_line
and	I-cell_line
non-lymphoid	I-cell_line
cells	I-cell_line
.	O

Here	O
we	O
establish	O
that	O
nuclear	O
extracts	O
from	O
distinct	O
cell	O
types	O
differ	O
in	O
their	O
ability	O
to	O
interact	O
with	O
octamer-containing	B-DNA
fragments	I-DNA
.	O

We	O
have	O
also	O
detected	O
a	O
DNA-protein	O
interaction	O
that	O
may	O
be	O
involved	O
in	O
the	O
cell-type	O
specificity	O
of	O
immunoglobulin	B-protein
expression	O
,	O
and	O
we	O
have	O
determined	O
that	O
a	O
sequence	O
upstream	O
of	O
the	O
octamer	O
participates	O
in	O
an	O
interaction	O
with	O
a	O
nuclear	B-protein
protein	I-protein
(	O
s	O
)	O
.	O

-DOCSTART-	O

Preferential	O
transcription	O
of	O
HTLV-I	B-DNA
LTR	I-DNA
in	O
cell-free	O
extracts	O
of	O
human	B-cell_type
T	I-cell_type
cells	I-cell_type
producing	O
HTLV-I	B-protein
viral	I-protein
proteins	I-protein
.	O

The	O
promoters	B-DNA
of	O
the	O
adenovirus	B-DNA
2	I-DNA
major	I-DNA
late	I-DNA
gene	I-DNA
,	O
the	O
mouse	B-DNA
beta-globin	I-DNA
gene	I-DNA
,	O
the	O
mouse	B-DNA
immunoglobulin	I-DNA
VH	I-DNA
gene	I-DNA
and	O
the	O
LTR	B-DNA
of	O
the	O
human	O
T-lymphotropic	O
retrovirus	O
type	O
I	O
were	O
tested	O
for	O
their	O
transcription	O
activities	O
in	O
cell-free	O
extracts	O
of	O
four	O
cell	O
lines	O
;	O
HeLa	B-cell_line
,	O
CESS	B-cell_line
(	O
Epstein-Barr	B-cell_line
virus-transformed	I-cell_line
human	I-cell_line
B	I-cell_line
cell	I-cell_line
line	I-cell_line
)	O
,	O
MT-1	B-cell_line
(	O
HTLV-I-infected	B-cell_line
human	I-cell_line
T	I-cell_line
cell	I-cell_line
line	I-cell_line
without	O
viral	O
protein	O
synthesis	O
)	O
,	O
and	O
MT-2	B-cell_line
(	O
HTLV-I-infected	B-cell_line
human	I-cell_line
T	I-cell_line
cell	I-cell_line
line	I-cell_line
producing	O
viral	B-protein
proteins	I-protein
)	O
.	O

LTR	B-DNA
was	O
preferentially	O
transcribed	O
in	O
the	O
extracts	O
of	O
MT-2	B-cell_line
although	O
the	O
other	O
three	O
genes	O
were	O
transcribed	O
with	O
relatively	O
constant	O
efficiencies	O
in	O
different	O
extracts	O
.	O

The	O
results	O
agree	O
well	O
with	O
the	O
previous	O
in	O
vivo	O
studies	O
on	O
the	O
promoter	O
activity	O
of	O
HTLV-I	B-DNA
LTR	I-DNA
.	O

Mixing	O
of	O
HeLa	O
and	O
MT-2	O
extracts	O
revealed	O
the	O
presence	O
of	O
a	O
LTR-specific	O
stimulating	O
activity	O
in	O
MT-2	O
extracts	O
.	O

-DOCSTART-	O

A	O
nuclear	B-protein
factor	I-protein
that	O
binds	O
to	O
a	O
conserved	O
sequence	O
motif	O
in	O
transcriptional	B-DNA
control	I-DNA
elements	I-DNA
of	O
immunoglobulin	B-DNA
genes	I-DNA
.	O

Trans-acting	B-protein
factors	I-protein
that	O
mediate	O
B-cell	O
specific	O
transcription	O
of	O
immunoglobulin	B-DNA
genes	I-DNA
have	O
been	O
postulated	O
based	O
on	O
an	O
analysis	O
of	O
the	O
expression	O
of	O
exogenously	O
introduced	O
immunoglobulin	B-DNA
gene	I-DNA
recombinants	I-DNA
in	O
lymphoid	B-cell_type
and	I-cell_type
non-lymphoid	I-cell_type
cells	I-cell_type
.	O

Two	O
B-cell-specific	O
,	O
cis-acting	B-DNA
transcriptional	I-DNA
regulatory	I-DNA
elements	I-DNA
have	O
been	O
identified	O
.	O

One	O
element	O
is	O
located	O
in	O
the	O
intron	O
between	O
the	O
variable	O
(	O
V	O
)	O
and	O
constant	O
(	O
C	O
)	O
regions	O
of	O
both	O
heavy	B-DNA
and	I-DNA
kappa	I-DNA
light-chain	I-DNA
genes	I-DNA
and	O
acts	O
as	O
a	O
transcriptional	O
enhancer	O
.	O

The	O
second	O
element	B-DNA
is	O
found	O
upstream	O
of	O
both	O
heavy	B-DNA
and	I-DNA
kappa	I-DNA
light-chain	I-DNA
gene	I-DNA
promoters	I-DNA
.	O

This	O
element	O
directs	O
lymphoid-specific	O
transcription	O
even	O
in	O
the	O
presence	O
of	O
viral	B-DNA
enhancers	I-DNA
.	O

We	O
have	O
sought	O
nuclear	O
factors	O
that	O
might	O
bind	O
specifically	O
to	O
these	O
two	O
regulatory	B-DNA
elements	I-DNA
by	O
application	O
of	O
a	O
modified	O
gel	O
electrophoresis	O
DNA	O
binding	O
assay	O
.	O

We	O
report	O
here	O
the	O
identification	O
of	O
a	O
human	B-protein
B-cell	I-protein
nuclear	I-protein
factor	I-protein
(	O
IgNF-A	B-protein
)	O
that	O
binds	O
to	O
DNA	B-DNA
sequences	I-DNA
in	O
the	O
upstream	B-DNA
regions	I-DNA
of	O
both	O
the	O
mouse	B-DNA
heavy	I-DNA
and	I-DNA
kappa	I-DNA
light-chain	I-DNA
gene	I-DNA
promoters	I-DNA
and	O
also	O
to	O
the	O
mouse	B-DNA
heavy-chain	I-DNA
gene	I-DNA
enhancer	I-DNA
.	O

This	O
sequence-specific	O
binding	O
is	O
probably	O
mediated	O
by	O
a	O
highly	O
conserved	O
sequence	O
motif	O
,	O
ATTTGCAT	O
,	O
present	O
in	O
all	O
three	O
transcriptional	B-DNA
elements	I-DNA
.	O

Interestingly	O
,	O
a	O
factor	O
showing	O
similar	O
binding	O
specificity	O
to	O
IgNF-A	B-DNA
is	O
also	O
present	O
in	O
human	B-cell_line
HeLa	I-cell_line
cells	I-cell_line
.	O

-DOCSTART-	O

The	O
new	O
world	O
primates	O
as	O
animal	O
models	O
of	O
glucocorticoid	O
resistance	O
.	O

Many	O
New	O
World	O
primate	O
species	O
have	O
greatly	O
increased	O
plasma	O
cortisol	O
concentrations	O
,	O
decreased	O
plasma	O
cortisol	O
binding	O
globulin	O
capacity	O
and	O
affinity	O
,	O
marked	O
resistance	O
of	O
the	O
hypothalamic-pituitary-adrenal	O
axis	O
to	O
suppression	O
by	O
dexamethasone	O
,	O
and	O
no	O
biological	O
evidence	O
of	O
glucocorticoid	O
excess	O
.	O

These	O
primates	O
also	O
have	O
high	O
levels	O
of	O
circulating	O
progesterone	O
,	O
estrogen	O
,	O
mineralocorticoid	O
,	O
androgen	O
and	O
vitamin	O
D	O
.	O

The	O
glucocorticoid	O
target	O
tissues	O
that	O
have	O
been	O
examined	O
(	O
circulating	B-cell_type
mononuclear	I-cell_type
lymphocytes	I-cell_type
and	O
cultured	B-cell_line
skin	I-cell_line
fibroblasts	I-cell_line
)	O
have	O
normal	O
concentrations	O
of	O
glucocorticoid	B-protein
receptors	I-protein
with	O
decreased	O
affinity	O
for	O
dexamethasone	O
.	O

Transformation	O
of	O
B-lymphocytes	B-cell_type
with	O
the	O
Epstein-Barr	O
virus	O
leads	O
to	O
glucocorticoid	B-protein
receptor	I-protein
induction	O
that	O
is	O
less	O
than	O
that	O
observed	O
with	O
cells	O
from	O
Old	O
World	O
primates	O
.	O

The	O
receptor	O
in	O
these	O
cells	O
has	O
a	O
low	O
affinity	O
for	O
dexamethasone	O
.	O

The	O
low	O
affinity	O
leads	O
to	O
an	O
increased	O
loss	O
of	O
specific	O
bound	O
ligand	O
during	O
thermal	O
activation	O
.	O

Meroreceptor	O
generation	O
is	O
normal	O
.	O

The	O
molecular	O
weight	O
of	O
the	O
receptor	O
,	O
determined	O
by	O
SDS-PAGE	O
,	O
is	O
similar	O
to	O
that	O
of	O
Old	O
World	O
primates	O
(	O
approximately	O
92	O
,	O
000	O
)	O
and	O
the	O
activation	O
pattern	O
per	O
se	O
,	O
examined	O
in	O
vitro	O
by	O
heating	O
cytosol	O
and	O
performing	O
phosphocellulose	O
chromatography	O
,	O
appears	O
similar	O
to	O
that	O
of	O
human	O
controls	O
.	O

The	O
ratios	O
of	O
nuclear	O
to	O
cytosolic	B-protein
hormone-receptor-complexes	I-protein
and	O
of	O
cytosolic	O
activated	O
to	O
unactivated	B-protein
receptor	I-protein
complexes	I-protein
in	O
intact	O
cells	O
are	O
similar	O
to	O
Old	O
World	O
primates	O
.	O

Results	O
from	O
mixing	O
studies	O
do	O
not	O
support	O
the	O
hypothesis	O
that	O
a	O
binding	O
inhibitor	O
(	O
s	O
)	O
or	O
a	O
deficient	O
cytosolic	O
positive	O
modifier	O
(	O
s	O
)	O
of	O
binding	O
underlies	O
the	O
findings	O
in	O
these	O
primates	O
.	O

The	O
New	O
World	O
primates	O
,	O
unlike	O
men	O
with	O
the	O
syndrome	O
of	O
primary	O
cortisol	O
resistance	O
,	O
have	O
compensated	O
for	O
their	O
condition	O
with	O
intra-adrenal	O
and	O
mineralocorticoid	O
receptor	O
adaptations	O
.	O

Thus	O
,	O
unlike	O
Old	O
World	O
primates	O
,	O
cortisol	O
in	O
New	O
World	O
primates	O
has	O
only	O
weak	O
sodium-retaining	O
potency	O
because	O
the	O
aldosterone	B-protein
receptor	I-protein
has	O
a	O
low	O
affinity	O
for	O
cortisol	O
.	O

The	O
common	O
element	O
that	O
would	O
explain	O
the	O
apparent	O
resistance	O
to	O
six	O
steroid	O
hormones	O
in	O
New	O
World	O
primates	O

-DOCSTART-	O

Acetylation	O
and	O
modulation	O
of	O
erythroid	B-protein
Kruppel-like	I-protein
factor	I-protein
(	O
EKLF	B-protein
)	O
activity	O
by	O
interaction	O
with	O
histone	B-protein
acetyltransferases	I-protein
.	O

Erythroid	B-protein
Kruppel-like	I-protein
factor	I-protein
(	O
EKLF	B-protein
)	O
is	O
a	O
red	B-protein
cell-specific	I-protein
transcriptional	I-protein
activator	I-protein
that	O
is	O
crucial	O
for	O
consolidating	O
the	O
switch	O
to	O
high	O
levels	O
of	O
adult	O
beta-globin	O
expression	O
during	O
erythroid	O
ontogeny	O
.	O

EKLF	B-protein
is	O
required	O
for	O
integrity	O
of	O
the	O
chromatin	O
structure	O
at	O
the	O
beta-like	B-DNA
globin	I-DNA
locus	I-DNA
,	O
and	O
it	O
interacts	O
with	O
a	O
positive-acting	B-protein
factor	I-protein
in	O
vivo	O
.	O

We	O
find	O
that	O
EKLF	B-protein
is	O
an	O
acetylated	B-protein
transcription	I-protein
factor	I-protein
,	O
and	O
that	O
it	O
interacts	O
in	O
vivo	O
with	O
CBP	B-protein
,	O
p300	B-protein
,	O
and	O
P/CAF	B-protein
.	O

However	O
,	O
its	O
interactions	O
with	O
these	O
histone	B-protein
acetyltransferases	I-protein
are	O
not	O
equivalent	O
,	O
as	O
CBP	B-protein
and	O
p300	B-protein
,	O
but	O
not	O
P/CAF	B-protein
,	O
utilize	O
EKLF	B-protein
as	O
a	O
substrate	O
for	O
in	O
vitro	O
acetylation	O
within	O
its	O
trans-activation	O
region	O
.	O

The	O
functional	O
effects	O
of	O
these	O
interactions	O
are	O
that	O
CBP	B-protein
and	O
p300	B-protein
,	O
but	O
not	O
P/CAF	B-protein
,	O
enhance	O
EKLF	B-protein
's	O
transcriptional	O
activation	O
of	O
the	O
beta-globin	B-DNA
promoter	I-DNA
in	O
erythroid	B-cell_type
cells	I-cell_type
.	O

These	O
results	O
establish	O
EKLF	B-protein
as	O
a	O
tissue-specific	B-protein
transcription	I-protein
factor	I-protein
that	O
undergoes	O
post-translational	O
acetylation	O
and	O
suggest	O
a	O
mechanism	O
by	O
which	O
EKLF	B-protein
is	O
able	O
to	O
alter	O
chromatin	O
structure	O
and	O
induce	O
beta-globin	O
expression	O
within	O
the	O
beta-like	B-DNA
globin	I-DNA
cluster	I-DNA
.	O

-DOCSTART-	O

Recognition	O
of	O
herpes	B-protein
simplex	I-protein
virus	I-protein
type	I-protein
2	I-protein
tegument	I-protein
proteins	I-protein
by	O
CD4	B-cell_type
T	I-cell_type
cells	I-cell_type
infiltrating	O
human	O
genital	O
herpes	O
lesions	O
.	O

The	O
local	O
cellular	O
immune	O
response	O
to	O
herpes	O
simplex	O
virus	O
(	O
HSV	O
)	O
is	O
important	O
in	O
the	O
control	O
of	O
recurrent	O
HSV	O
infection	O
.	O

The	O
antiviral	O
functions	O
of	O
infiltrating	B-cell_type
CD4-bearing	I-cell_type
T	I-cell_type
cells	I-cell_type
may	O
include	O
cytotoxicity	O
,	O
inhibition	O
of	O
viral	O
growth	O
,	O
lymphokine	O
secretion	O
,	O
and	O
support	O
of	O
humoral	O
and	O
CD8	O
responses	O
.	O

The	O
antigens	O
recognized	O
by	O
many	O
HSV-specific	B-cell_type
CD4	I-cell_type
T	I-cell_type
cells	I-cell_type
localizing	O
to	O
genital	O
HSV-2	O
lesions	O
are	O
unknown	O
.	O

T	B-cell_type
cells	I-cell_type
recognizing	O
antigens	O
encoded	O
within	O
map	O
units	O
0.67	O
to	O
0.73	O
of	O
HSV	B-DNA
DNA	I-DNA
are	O
frequently	O
recovered	O
from	O
herpetic	O
lesions	O
.	O

Expression	O
cloning	O
with	O
this	O
region	O
of	O
DNA	O
now	O
shows	O
that	O
tegument	B-protein
protein	I-protein
VP22	B-protein
and	O
the	O
viral	B-protein
dUTPase	I-protein
,	O
encoded	O
by	O
genes	B-DNA
UL49	I-DNA
and	I-DNA
UL50	I-DNA
,	O
respectively	O
,	O
are	O
T-cell	B-protein
antigens	I-protein
.	O

Separate	O
epitopes	B-protein
in	O
VP22	B-protein
were	O
defined	O
for	O
T-cell	B-cell_line
clones	I-cell_line
from	O
each	O
of	O
three	O
patients	O
.	O

Reactivity	O
with	O
the	O
tegument	B-protein
protein	I-protein
encoded	O
by	O
UL21	B-DNA
was	O
identified	O
for	O
an	O
additional	O
patient	O
.	O

Three	O
new	O
epitopes	O
were	O
identified	O
in	O
VP16	B-protein
,	O
a	O
tegument	B-protein
protein	I-protein
associated	O
with	O
VP22	B-protein
.	O

Some	O
tegument-specific	B-cell_line
CD4	I-cell_line
T-cell	I-cell_line
clones	I-cell_line
exhibited	O
cytotoxic	O
activity	O
against	O
HSV-infected	B-cell_type
cells	I-cell_type
.	O

These	O
results	O
suggest	O
that	O
herpes	B-protein
simplex	I-protein
tegument	I-protein
proteins	I-protein
are	O
processed	O
for	O
antigen	O
presentation	O
in	O
vivo	O
and	O
are	O
possible	O
candidate	O
compounds	O
for	O
herpes	O
simplex	O
vaccines	O
.	O

-DOCSTART-	O

Fibrinogen	B-protein
activates	O
NF-kappa	B-protein
B	I-protein
transcription	I-protein
factors	I-protein
in	O
mononuclear	B-cell_type
phagocytes	I-cell_type
.	O

Adhesion	O
to	O
extracellular	O
matrices	O
is	O
known	O
to	O
modulate	O
leukocyte	O
activation	O
,	O
although	O
the	O
mechanisms	O
are	O
not	O
fully	O
understood	O
.	O

Mononuclear	B-cell_type
phagocytes	I-cell_type
are	O
exposed	O
to	O
fibrinous	O
provisional	O
matrix	O
throughout	O
migration	O
into	O
inflammatory	O
foci	O
,	O
so	O
this	O
study	O
was	O
undertaken	O
to	O
determine	O
whether	O
fibrinogen	O
triggers	O
activation	O
of	O
NF-kappa	B-protein
B	I-protein
transcription	I-protein
factors	I-protein
.	O

U937	B-cell_line
cells	I-cell_line
differentiated	O
with	O
PMA	O
in	O
nonadherent	B-cell_line
culture	I-cell_line
were	O
shown	O
to	O
express	O
two	O
fibrinogen-binding	B-protein
integrins	I-protein
,	O
predominately	O
CD11b/CD18	B-protein
,	O
and	O
to	O
a	O
lesser	O
extent	O
,	O
CD11c/CD18	B-protein
.	O

Cells	O
stimulated	O
with	O
fibrinogen	B-protein
(	O
10-100	O
microg/ml	O
)	O
/Mn2+	O
(	O
50	O
microM	O
)	O
for	O
2	O
h	O
were	O
examined	O
by	O
electrophoretic	O
mobility	O
shift	O
assay	O
.	O

NF-kappa	B-protein
B	I-protein
activation	O
,	O
minimal	O
in	O
unstimulated	B-cell_type
cells	I-cell_type
,	O
was	O
substantially	O
up-regulated	O
by	O
fibrinogen	B-protein
.	O

Fibrinogen	B-protein
also	O
caused	O
activation	O
of	O
AP-1	B-protein
,	O
but	O
not	O
SP1	B-protein
or	O
cAMP	B-protein
response	I-protein
element-binding	I-protein
protein	I-protein
(	I-protein
CREB	I-protein
)	I-protein
factors	I-protein
.	O

Blocking	O
mAbs	B-protein
against	O
CD18	B-protein
and	O
CD11b	B-protein
abrogated	O
fibrinogen	B-protein
-induced	O
NF-kappa	B-protein
B	I-protein
activation	O
.	O

To	O
determine	O
the	O
effects	O
on	O
transcriptional	O
regulation	O
,	O
U937	B-cell_line
cells	I-cell_line
were	O
transfected	O
with	O
a	O
plasmid	O
containing	O
the	O
HIV-1	B-DNA
enhancer	I-DNA
(	O
bearing	O
two	O
NF-kappa	B-DNA
B	I-DNA
sites	I-DNA
)	O
coupled	O
to	O
a	O
chloramphenicol	B-DNA
acetyltransferase	I-DNA
(	I-DNA
CAT	I-DNA
)	I-DNA
reporter	I-DNA
.	O

Cells	O
were	O
subsequently	O
stimulated	O
with	O
1	O
)	O
PMA	O
for	O
24	O
h	O
,	O
inducing	O
CAT	O
activity	O
by	O
2.6-fold	O
,	O
2	O
)	O
fibrinogen	B-protein
/Mn2+	O
for	O
2	O
h	O
,	O
inducing	O
CAT	O
activity	O
by	O
3.2-fold	O
,	O
or	O
3	O
)	O
costimulation	O
with	O
fibrinogen	B-protein
and	O
PMA	O
,	O
inducing	O
5.7-fold	O
the	O
CAT	O
activity	O
induced	O
by	O
PMA	O
alone	O
.	O

We	O
conclude	O
that	O
contact	O
with	O
fibrinogen-derived	B-protein
proteins	I-protein
may	O
contribute	O
to	O
mononuclear	O
phagocyte	O
activation	O
by	O
signaling	O
through	O
CD11b/CD18	B-protein
,	O
resulting	O
in	O
selective	O
activation	O
of	O
transcriptional	B-protein
regulatory	I-protein
factors	I-protein
,	O
including	O
NF-kappa	B-protein
B	I-protein
.	O

-DOCSTART-	O

Peripheral	B-cell_type
blood	I-cell_type
T	I-cell_type
cells	I-cell_type
and	I-cell_type
monocytes	I-cell_type
and	O
B	B-cell_line
cell	I-cell_line
lines	I-cell_line
derived	O
from	O
patients	O
with	O
lupus	O
express	O
estrogen	O
receptor	O
transcripts	O
similar	O
to	O
those	O
of	O
normal	B-cell_type
cells	I-cell_type
.	O

OBJECTIVE	O
:	O
To	O
identify	O
and	O
characterize	O
estrogen	B-RNA
receptor	I-RNA
(	I-RNA
ER	I-RNA
)	I-RNA
transcripts	I-RNA
expressed	O
in	O
immune	B-cell_type
cells	I-cell_type
of	O
patients	O
with	O
systemic	O
lupus	O
erythematosus	O
(	O
SLE	O
)	O
and	O
healthy	O
donors	O
.	O

METHODS	O
:	O
Peripheral	B-cell_type
blood	I-cell_type
monocytes	I-cell_type
and	I-cell_type
T	I-cell_type
cells	I-cell_type
were	O
prepared	O
from	O
patients	O
with	O
SLE	O
(	O
n	O
=	O
6	O
)	O
and	O
healthy	O
donors	O
(	O
n	O
=	O
8	O
)	O
.	O

T	B-cell_type
cells	I-cell_type
were	O
separated	O
into	O
CD4	B-cell_type
and	O
CD8	B-cell_type
.	O

Some	O
monocytes	B-cell_type
and	O
T	B-cell_type
cells	I-cell_type
were	O
stimulated	O
with	O
estradiol	O
,	O
PMA	O
,	O
and	O
ionomycin	O
.	O

Epstein-Barr	O
virus-transformed	O
B	B-cell_line
cell	I-cell_line
lines	I-cell_line
(	O
n	O
=	O
7	O
)	O
and	O
B	B-cell_line
cell	I-cell_line
hybridomas	I-cell_line
(	O
n	O
=	O
2	O
)	O
established	O
from	O
patients	O
with	O
SLE	O
and	O
a	O
healthy	O
individual	O
were	O
used	O
as	O
a	O
B	O
cell	O
source	O
.	O

These	O
cells	O
were	O
examined	O
for	O
ER	B-RNA
mRNA	I-RNA
by	O
reverse	O
transcription	O
nested	O
polymerase	O
chain	O
reaction	O
.	O

Amplified	O
cDNA	B-DNA
were	O
sequenced	O
by	O
standard	O
methods	O
.	O

RESULTS	O
:	O
In	O
all	O
cells	O
tested	O
,	O
ER	B-RNA
mRNA	I-RNA
was	O
expressed	O
without	O
prior	O
in	O
vitro	O
stimulation	O
.	O

Partial	O
sequences	O
from	O
exons	B-DNA
1-8	I-DNA
were	O
nearly	O
identical	O
to	O
the	O
published	O
sequence	O
of	O
the	O
human	B-RNA
ER	I-RNA
mRNA	I-RNA
.	O

There	O
were	O
no	O
notable	O
differences	O
in	O
the	O
ER	B-RNA
transcripts	I-RNA
between	O
patients	O
and	O
healthy	O
controls	O
.	O

Variant	O
receptor	O
transcripts	O
lacking	O
exon	B-DNA
5	I-DNA
or	O
exon	B-DNA
7	I-DNA
,	O
which	O
encodes	O
the	O
hormone	B-protein
binding	I-protein
domain	I-protein
,	O
were	O
identified	O
in	O
the	O
majority	O
of	O
the	O
cells	O
.	O

Precise	O
deletion	O
of	O
the	O
exons	O
suggests	O
that	O
they	O
are	O
alternatively	O
spliced	O
transcripts	O
.	O

Whether	O
the	O
detected	O
transcripts	O
are	O
translated	O
into	O
functional	B-protein
receptor	I-protein
proteins	I-protein
remains	O
to	O
be	O
determined	O
.	O

In	O
vitro	O
stimulation	O
did	O
not	O
affect	O
ER	B-RNA
mRNA	I-RNA
expression	O
.	O

The	O
presence	O
of	O
variants	O
did	O
not	O
correlate	O
with	O
disease	O
activity	O
or	O
medication	O
.	O

CONCLUSION	O
:	O
Monocytes	B-cell_type
,	O
T	B-cell_type
cells	I-cell_type
,	O
and	O
B	B-cell_type
cells	I-cell_type
in	O
patients	O
express	O
transcripts	O
of	O
the	O
normal	O
wild	B-protein
type	I-protein
ER	I-protein
and	O
the	O
hormone	B-protein
binding	I-protein
domain	I-protein
variants	I-protein
in	O
vivo	O
.	O

-DOCSTART-	O

DNA	O
damaging	O
agents	O
induce	O
expression	O
of	O
Fas	B-protein
ligand	I-protein
and	O
subsequent	O
apoptosis	O
in	O
T	B-cell_type
lymphocytes	I-cell_type
via	O
the	O
activation	O
of	O
NF-kappa	B-protein
B	I-protein
and	O
AP-1	B-protein
.	O

Apoptosis	O
induced	O
by	O
DNA	O
damage	O
and	O
other	O
stresses	O
can	O
proceed	O
via	O
expression	O
of	O
Fas	B-protein
ligand	I-protein
(	O
FasL	B-protein
)	O
and	O
ligation	O
of	O
its	O
receptor	O
,	O
Fas	O
(	O
CD95	O
)	O
.	O

We	O
report	O
that	O
activation	O
of	O
the	O
two	O
transcription	B-protein
factors	I-protein
NF-kappa	I-protein
B	I-protein
and	I-protein
AP-1	I-protein
is	O
crucially	O
involved	O
in	O
FasL	B-protein
expression	O
induced	O
by	O
etoposide	O
,	O
teniposide	O
,	O
and	O
UV	O
irradiation	O
.	O

A	O
nondegradable	B-protein
mutant	I-protein
of	O
I	B-protein
kappa	I-protein
B	I-protein
blocked	O
both	O
FasL	B-protein
expression	O
and	O
apoptosis	O
induced	O
by	O
DNA	O
damage	O
but	O
not	O
Fas	O
ligation	O
.	O

These	O
stimuli	O
also	O
induced	O
the	O
stress-activated	O
kinase	O
pathway	O
(	O
SAPK/JNK	O
)	O
,	O
which	O
was	O
required	O
for	O
the	O
maximal	O
induction	O
of	O
apoptosis	O
.	O

A	O
1.2	O
kb	O
FasL	B-DNA
promoter	I-DNA
responded	O
to	O
DNA	O
damage	O
,	O
as	O
well	O
as	O
coexpression	O
with	O
p65	B-protein
Rel	I-protein
or	O
Fos/Jun	B-protein
.	O

Mutations	O
in	O
the	O
relevant	O
NF-kappa	B-DNA
B	I-DNA
and	I-DNA
AP-1	I-DNA
binding	I-DNA
sites	I-DNA
eliminated	O
these	O
responses	O
.	O

Thus	O
,	O
activation	O
of	O
NF-kappa	B-protein
B	I-protein
and	O
AP-1	B-protein
contributes	O
to	O
stress-induced	O
apoptosis	O
via	O
the	O
expression	O
of	O
FasL	B-protein
.	O

-DOCSTART-	O

A	O
small	O
,	O
nonpeptidyl	O
mimic	O
of	O
granulocyte-colony-stimulating	B-protein
factor	I-protein
[	O
see	O
commetns	O
]	O

A	O
nonpeptidyl	O
small	O
molecule	O
SB	O
247464	O
,	O
capable	O
of	O
activating	O
granulocyte-colony-stimulating	B-protein
factor	I-protein
(	O
G-CSF	B-protein
)	O
signal	O
transduction	O
pathways	O
,	O
was	O
identified	O
in	O
a	O
high-throughput	O
assay	O
in	O
cultured	B-cell_type
cells	I-cell_type
.	O

Like	O
G-CSF	B-protein
,	O
SB	O
247464	O
induced	O
tyrosine	O
phosphorylation	O
of	O
multiple	B-protein
signaling	I-protein
proteins	I-protein
and	O
stimulated	O
primary	B-cell_type
murine	I-cell_type
bone	I-cell_type
marrow	I-cell_type
cells	I-cell_type
to	O
form	O
granulocytic	B-cell_type
colonies	I-cell_type
in	O
vitro	O
.	O

It	O
also	O
elevated	O
peripheral	B-cell_type
blood	I-cell_type
neutrophil	I-cell_type
counts	O
in	O
mice	O
.	O

The	O
extracellular	O
domain	O
of	O
the	O
murine	B-protein
G-CSF	I-protein
receptor	I-protein
was	O
required	O
for	O
the	O
activity	O
of	O
SB	O
247464	O
,	O
suggesting	O
that	O
the	O
compound	O
acts	O
by	O
oligomerizing	B-protein
receptor	I-protein
chains	I-protein
.	O

The	O
results	O
indicate	O
that	O
a	O
small	O
molecule	O
can	O
activate	O
a	O
receptor	O
that	O
normally	O
binds	O
a	O
relatively	O
large	O
protein	O
ligand	O
.	O

-DOCSTART-	O

Minimal	O
residual	O
disease	O
in	O
acute	O
myelogenous	O
leukemia	O
with	O
PML/RAR	B-protein
alpha	I-protein
or	O
AML1/ETO	B-RNA
mRNA	I-RNA
and	O
phenotypic	O
analysis	O
of	O
possible	O
T	B-cell_type
and	I-cell_type
natural	I-cell_type
killer	I-cell_type
cells	I-cell_type
in	O
bone	O
marrow	O
.	O

Here	O
we	O
studied	O
minimal	O
residual	O
disease	O
(	O
MRD	O
)	O
of	O
patients	O
with	O
acute	O
myeloid	O
leukemia	O
(	O
AML	O
)	O
who	O
have	O
PML/RAR	B-protein
alpha	I-protein
or	O
AML1/ETO	B-protein
as	O
well	O
as	O
the	O
phenotypic	O
analysis	O
of	O
lymphocyte	B-cell_type
subsets	I-cell_type
involved	O
in	O
antitumor	O
immunity	O
.	O

Eight	O
patients	O
in	O
long-term	O
(	O
LT	O
;	O
3	O
to	O
15	O
years	O
)	O
and	O
15	O
patients	O
in	O
short-term	O
(	O
ST	O
;	O
up	O
to	O
3	O
years	O
)	O
remission	O
were	O
studied	O
.	O

Using	O
the	O
reverse	O
transcription-polymerase	O
chain	O
reaction	O
(	O
RT	O
)	O
assay	O
,	O
the	O
limit	O
of	O
detection	O
was	O
10	O
(	O
-5	O
)	O
to	O
10	O
(	O
-6	O
)	O
for	O
PML/RAR	B-RNA
alpha	I-RNA
transcript	I-RNA
and	O
10	O
(	O
-4	O
)	O
to	O
10	O
(	O
-5	O
)	O
for	O
the	O
AML1/ETO	B-RNA
transcript	I-RNA
.	O

Simultaneously	O
,	O
T	B-cell_type
lymphocyte	I-cell_type
subsets	I-cell_type
and	O
NK	B-cell_type
cells	I-cell_type
from	O
the	O
peripheral	O
blood	O
(	O
PB	O
)	O
and	O
bone	O
marrow	O
(	O
BM	O
)	O
were	O
investigated	O
by	O
flow	O
cytometric	O
analysis	O
.	O

Four	O
of	O
the	O
eight	O
patients	O
in	O
LT	O
and	O
7	O
of	O
the	O
15	O
patients	O
in	O
ST	O
remission	O
were	O
MRD-positive	O
.	O

Although	O
all	O
MRD-positive	O
patients	O
in	O
LT	O
remission	O
are	O
still	O
until	O
now	O
event-free	O
,	O
3	O
of	O
the	O
7	O
MRD-positive	O
(	O
MRD+	O
)	O
patients	O
in	O
ST	O
remission	O
soon	O
relapsed	O
.	O

The	O
total	O
populations	O
of	O
CD4	B-protein
+	O
,	O
CD8	B-protein
+	O
and	O
CD56	B-protein
+	O
[	O
possible	O
T-cell	B-cell_type
and	I-cell_type
natural	I-cell_type
killer	I-cell_type
(	I-cell_type
T/NK	I-cell_type
)	I-cell_type
populations	I-cell_type
]	O
in	O
the	O
BM	O
of	O
ST	O
patients	O
and	O
MRD+/LT	O
patients	O
were	O
significantly	O
(	O
p	O
<	O
.01	O
)	O
low	O
.	O

The	O
CD8+	B-cell_type
CD28+	I-cell_type
population	I-cell_type
showed	O
the	O
same	O
tendency	O
(	O
p	O
<	O
.01-.02	O
)	O
.	O

The	O
T/NK	B-cell_type
subsets	I-cell_type
in	O
the	O
BM	O
of	O
MRD-negative	O
(	O
MRD-	O
)	O
LT	O
(	O
MRD-/LT	O
)	O
patients	O
showed	O
similar	O
numbers	O
of	O
cells	O
as	O
normal	O
volunteers	O
.	O

Basically	O
,	O
the	O
total	O
percentage	O
of	O
the	O
CD4+	B-cell_type
,	I-cell_type
CD8+	I-cell_type
and	I-cell_type
CD56+	I-cell_type
cell	I-cell_type
populations	I-cell_type
in	O
the	O
BM	O
was	O
increased	O
and	O
in	O
the	O
following	O
order	O
:	O
MRD-/LT	O
patients	O
,	O
normal	O
volunteers	O
,	O
MRD+/LT	O
patients	O
and	O
MRD+	O
or	O
-/ST	O
patients	O
.	O

The	O
percentages	O
of	O
the	O
T/NK-cell	B-cell_type
subsets	I-cell_type
in	O
the	O
PB	O
were	O
not	O
significantly	O
different	O
among	O
these	O
groups	O
.	O

Thus	O
,	O
the	O
difference	O
of	O
the	O
possible	O
T/NK-cell	B-cell_type
phenotype	O
in	O
the	O
BM	O
may	O
strongly	O
influence	O
clinical	O
and	O
molecular	O
remission	O
.	O

These	O
results	O
still	O
remain	O
to	O
be	O
confirmed	O
by	O
further	O
studies	O
of	O
the	O
functional	O
anti-tumor	O
immunity	O
of	O
T/NK	B-cell_type
cells	I-cell_type
of	O
AML	B-cell_type
in	O
remission	O
.	O

-DOCSTART-	O

Mycobacterium	O
tuberculosis	O
mannose-capped	O
lipoarabinomannan	O
can	O
induce	O
NF-kappaB	B-protein
-dependent	O
activation	O
of	O
human	B-DNA
immunodeficiency	I-DNA
virus	I-DNA
type	I-DNA
1	I-DNA
long	I-DNA
terminal	I-DNA
repeat	I-DNA
in	O
T	B-cell_type
cells	I-cell_type
.	O

Tuberculosis	O
has	O
emerged	O
as	O
an	O
epidemic	O
,	O
extended	O
by	O
the	O
large	O
number	O
of	O
individuals	O
infected	O
with	O
human	O
immunodeficiency	O
virus	O
type	O
1	O
(	O
HIV-1	O
)	O
.	O

The	O
major	O
goal	O
of	O
this	O
study	O
was	O
to	O
determine	O
whether	O
the	O
mycobacterial	O
cell	O
wall	O
component	O
mannose-capped	O
lipoarabinomannan	O
(	O
ManLAM	O
)	O
of	O
Mycobacterium	O
tuberculosis	O
(	O
M.	O
tuberculosis	O
)	O
could	O
activate	O
transcription	O
of	O
HIV-1	O
in	O
T	B-cell_type
cells	I-cell_type
with	O
the	O
use	O
of	O
an	O
in	O
vitro	O
cell	O
culture	O
system	O
.	O

These	O
experiments	O
are	O
of	O
prime	O
importance	O
considering	O
that	O
CD4	B-protein
-expressing	O
T	B-cell_type
lymphocytes	I-cell_type
represent	O
the	O
major	O
virus	O
reservoir	O
in	O
the	O
peripheral	O
blood	O
of	O
infected	O
individuals	O
.	O

Using	O
the	O
1G5	B-cell_line
cell	I-cell_line
line	I-cell_line
harbouring	O
the	O
luciferase	O
reporter	O
gene	O
under	O
the	O
control	O
of	O
the	O
HIV-1	B-DNA
LTR	I-DNA
,	O
it	O
was	O
first	O
found	O
that	O
culture	O
protein	O
filtrates	O
(	O
CFP	O
)	O
from	O
M.	O
tuberculosis	O
or	O
purified	O
ManLAM	O
could	O
activate	O
HIV-1	B-DNA
LTR-dependent	I-DNA
gene	I-DNA
expression	O
unlike	O
similarly	O
prepared	O
CFP	O
extracts	O
devoid	O
of	O
ManLAM	O
.	O

The	O
implication	O
of	O
protein	B-protein
tyrosine	I-protein
kinase	I-protein
(	I-protein
s	I-protein
)	I-protein
,	O
protein	B-protein
kinase	I-protein
A	I-protein
and/or	O
protein	B-protein
kinase	I-protein
C	I-protein
was	O
highlighted	O
by	O
the	O
abrogation	O
of	O
the	O
ManLAM-mediated	O
activation	O
of	O
HIV-1	O
LTR-driven	O
gene	O
expression	O
using	O
herbimycin	O
A	O
and	O
H7	O
.	O

It	O
was	O
also	O
determined	O
,	O
using	O
electrophoresis	O
mobility	O
shift	O
assays	O
,	O
that	O
M.	O
tuberculosis	O
ManLAM	O
led	O
to	O
the	O
nuclear	O
translocation	O
of	O
the	O
transcription	B-protein
factor	I-protein
NF-kappaB	I-protein
.	O

M.	O
tuberculosis	O
ManLAM	O
resulted	O
in	O
clear	O
induction	O
of	O
the	O
luciferase	B-DNA
gene	I-DNA
placed	O
under	O
the	O
control	O
of	O
the	O
wild-type	B-DNA
,	I-DNA
but	I-DNA
not	I-DNA
the	I-DNA
kappaB-mutated	I-DNA
,	I-DNA
HIV-1	I-DNA
LTR	I-DNA
region	I-DNA
.	O

Finally	O
,	O
the	O
ManLAM-mediated	O
activation	O
of	O
HIV-1	O
LTR	O
transcription	O
was	O
found	O
to	O
be	O
independent	O
of	O
the	O
autocrine	O
or	O
paracrine	O
action	O
of	O
endogenous	B-protein
TNF-alpha	I-protein
.	O

The	O
results	O
suggest	O
that	O
M.	O
tuberculosis	O
can	O
upregulate	O
HIV-1	O
expression	O
in	O
T	B-cell_type
cells	I-cell_type
and	O
could	O
thus	O
have	O
the	O
potential	O
to	O
influence	O
the	O
pathogenesis	O
of	O
HIV-1	O
infection	O
.	O

-DOCSTART-	O

Human	B-DNA
immunodeficiency	I-DNA
virus	I-DNA
type	I-DNA
1	I-DNA
long	I-DNA
terminal	I-DNA
repeat	I-DNA
quasispecies	O
differ	O
in	O
basal	O
transcription	O
and	O
nuclear	O
factor	O
recruitment	O
in	O
human	B-cell_type
glial	I-cell_type
cells	I-cell_type
and	I-cell_type
lymphocytes	I-cell_type
.	O

The	O
generation	O
of	O
genomic	O
diversity	O
during	O
the	O
course	O
of	O
infection	O
has	O
the	O
potential	O
to	O
affect	O
all	O
aspects	O
of	O
HIV-1	O
replication	O
,	O
including	O
expression	O
of	O
the	O
proviral	O
genome	O
.	O

To	O
gain	O
a	O
better	O
understanding	O
of	O
the	O
impact	O
of	O
long	B-DNA
terminal	I-DNA
repeat	I-DNA
(	O
LTR	B-DNA
)	O
sequence	O
diversity	O
on	O
LTR-directed	B-DNA
gene	I-DNA
expression	O
in	O
cells	O
of	O
the	O
central	O
nervous	O
system	O
(	O
CNS	O
)	O
and	O
immune	O
system	O
,	O
we	O
amplified	O
and	O
cloned	O
LTRs	B-DNA
from	O
proviral	O
DNA	O
in	O
HIV-1-infected	O
peripheral	O
blood	O
.	O

Sequence	O
analysis	O
of	O
nineteen	O
LTRs	B-DNA
cloned	O
from	O
2	O
adult	O
and	O
3	O
pediatric	O
patients	O
revealed	O
an	O
average	O
of	O
33	O
nucleotide	O
changes	O
(	O
with	O
respect	O
to	O
the	O
sequence	O
of	O
the	O
LAI	B-DNA
LTR	I-DNA
)	O
within	O
the	O
455-bp	B-DNA
U3	I-DNA
region	I-DNA
.	O

Transient	O
expression	O
analyses	O
in	O
cells	O
of	O
neuroglial	O
and	O
lymphocytic	O
origin	O
demonstrated	O
that	O
some	O
of	O
these	O
LTRs	B-DNA
had	O
activities	O
which	O
varied	O
significantly	O
from	O
the	O
LAI	B-DNA
LTR	I-DNA
in	O
U-373	B-cell_line
MG	I-cell_line
cells	I-cell_line
(	O
an	O
astrocytoma	B-cell_line
cell	I-cell_line
line	I-cell_line
)	O
as	O
well	O
as	O
in	O
Jurkat	B-cell_line
cells	I-cell_line
(	O
a	O
CD4-positive	B-cell_line
lymphocyte	I-cell_line
cell	I-cell_line
line	I-cell_line
)	O
.	O

While	O
LTRs	B-DNA
which	O
demonstrated	O
the	O
highest	O
activities	O
in	O
U-373	B-cell_line
MG	I-cell_line
cells	I-cell_line
also	O
yielded	O
high	O
activities	O
in	O
Jurkat	B-cell_line
cells	I-cell_line
,	O
the	O
LTRs	B-DNA
were	O
generally	O
more	O
active	O
in	O
Jurkat	B-cell_line
cells	I-cell_line
when	O
compared	O
to	O
the	O
LAI	B-DNA
LTR	I-DNA
.	O

Differences	O
in	O
LTR	B-DNA
sequence	I-DNA
also	O
resulted	O
in	O
differences	O
in	O
transcription	O
factor	O
recruitment	O
to	O
cis-acting	B-DNA
sites	I-DNA
within	O
the	O
U3	B-DNA
region	I-DNA
of	O
the	O
LTR	B-DNA
,	O
as	O
demonstrated	O
by	O
electrophoretic	O
mobility	O
shift	O
assays	O
.	O

In	O
particular	O
,	O
naturally	O
occurring	O
sequence	O
variation	O
impacted	O
transcription	B-protein
factor	I-protein
binding	O
to	O
an	O
activating	B-protein
transcription	I-protein
factor	I-protein
/	O
cAMP	B-protein
response	I-protein
element	I-protein
binding	I-protein
(	O
ATF	B-protein
/	O
CREB	B-protein
)	O
binding	O
site	O
(	O
located	O
between	O
the	O
LEF-1	B-DNA
and	I-DNA
distal	I-DNA
NF-kappaB	I-DNA
transcription	I-DNA
factor	I-DNA
binding	I-DNA
sites	I-DNA
)	O
that	O
we	O
identified	O
in	O
previous	O
studies	O
of	O
the	O
HIV-1	B-DNA
LTR	I-DNA
.	O

These	O
findings	O
suggest	O
that	O
LTR	B-DNA
sequence	I-DNA
changes	O
can	O
significantly	O
affect	O
basal	O
LTR	O
function	O
and	O
transcription	O
factor	O
recruitment	O
,	O
which	O
may	O
,	O
in	O
turn	O
,	O
alter	O
the	O
course	O
of	O
viral	O
replication	O
in	O
cells	O
of	O
CNS	O
and	O
immune	O
system	O
origin	O
.	O

-DOCSTART-	O

HMG	B-protein
box	I-protein
containing	I-protein
transcription	I-protein
factors	I-protein
in	O
lymphocyte	B-cell_type
differentiation	O
.	O

The	O
identification	O
of	O
the	O
mammalian	B-DNA
sex-determining	I-DNA
gene	I-DNA
Sry	I-DNA
has	O
led	O
to	O
the	O
discovery	O
of	O
a	O
large	O
family	O
of	O
related	O
(	O
'	O
HMG	B-protein
box	I-protein
'	O
)	O
transcription	B-protein
factors	I-protein
that	O
control	O
developmental	O
events	O
in	O
yeast	O
,	O
C.	O
elegans	O
,	O
Drosophila	O
and	O
vertebrates	O
.	O

In	O
lymphocyte	B-cell_type
differentiation	O
,	O
several	O
HMG	B-protein
box	I-protein
proteins	I-protein
play	O
a	O
decisive	O
role	O
.	O

Sox-4	B-protein
is	O
important	O
for	O
very	O
early	O
B-cell	O
differentiation	O
,	O
while	O
TCF-1	B-protein
/	O
LEF-1	B-protein
play	O
a	O
crucial	O
role	O
in	O
early	O
thymocyte	O
development	O
.	O

TCF/LEF	B-protein
proteins	I-protein
have	O
recently	O
been	O
found	O
to	O
constitute	O
a	O
downstream	O
component	O
of	O
the	O
Wingless/Wnt	O
signal	O
transduction	O
pathway	O
.	O

In	O
flies	O
,	O
this	O
pathway	O
controls	O
segment	O
polarity	O
;	O
in	O
Xenopus	O
it	O
controls	O
the	O
definition	O
of	O
the	O
body	O
axis	O
.	O

Deregulation	O
of	O
the	O
pathway	O
occurs	O
in	O
several	O
human	B-cell_type
tumors	I-cell_type
.	O

These	O
insights	O
in	O
the	O
molecular	O
events	O
that	O
are	O
involved	O
in	O
TCF/LEF	B-protein
function	O
in	O
these	O
organisms	O
may	O
eventually	O
lead	O
to	O
the	O
understanding	O
of	O
the	O
function	O
of	O
these	O
HMG	B-protein
box	I-protein
proteins	I-protein
in	O
lymphoid	O
development	O

-DOCSTART-	O

Transcriptional	O
regulation	O
by	O
C/EBP	B-protein
alpha	I-protein
and	I-protein
-beta	I-protein
in	O
the	O
expression	O
of	O
the	O
gene	O
for	O
the	O
MRP14	B-protein
myeloid	I-protein
calcium	I-protein
binding	I-protein
protein	I-protein
.	O

Transcriptional	O
regulation	O
of	O
the	O
gene	O
for	O
the	O
myeloid	B-protein
calcium	I-protein
binding	I-protein
protein	I-protein
,	O
MRP14	B-protein
,	O
was	O
investigated	O
in	O
human	B-cell_line
monocytic	I-cell_line
leukemia	I-cell_line
cell	I-cell_line
lines	I-cell_line
.	O

The	O
MRP14	B-DNA
gene	I-DNA
was	O
not	O
expressed	O
in	O
monoblastic	B-cell_line
ML-1	I-cell_line
cells	I-cell_line
,	O
promonocytic	B-cell_line
U-937	I-cell_line
cells	I-cell_line
,	O
or	O
promyelocytic	B-cell_line
HL-60	I-cell_line
cells	I-cell_line
.	O

On	O
the	O
other	O
hand	O
,	O
the	O
gene	O
was	O
expressed	O
in	O
monocytic	B-cell_line
THP-1	I-cell_line
cells	I-cell_line
and	O
in	O
the	O
HL-60	B-cell_line
cells	I-cell_line
treated	O
with	O
1	O
,	O
25-dihydroxyvitamin	O
D3	O
(	O
VD3	O
)	O
.	O

The	O
level	O
of	O
MRP14	B-protein
in	O
VD3-treated	O
HL-60	B-cell_line
cells	I-cell_line
was	O
two-fold	O
higher	O
than	O
that	O
in	O
THP-1	B-cell_line
cells	I-cell_line
.	O

Among	O
several	O
known	O
transcription	B-DNA
factor	I-DNA
binding	I-DNA
motifs	I-DNA
,	O
nuclear	B-protein
protein	I-protein
(	I-protein
s	I-protein
)	I-protein
of	O
VD3-treated	B-cell_line
HL-60	I-cell_line
cells	I-cell_line
and	I-cell_line
THP-1	I-cell_line
cells	I-cell_line
bound	O
to	O
the	O
CCAAT/enhancer	B-DNA
binding	I-DNA
protein	I-DNA
(	I-DNA
C/EBP	I-DNA
)	I-DNA
-binding	I-DNA
motif	I-DNA
that	O
was	O
located	O
in	O
the	O
upstream	B-DNA
region	I-DNA
of	O
the	O
MRP14	B-DNA
gene	I-DNA
(	O
-81	O
)	O
,	O
as	O
evidenced	O
by	O
the	O
competitive	O
gel	O
mobility-shift	O
assay	O
.	O

An	O
antibody	O
for	O
C/EBP	B-protein
alpha	I-protein
super-shifted	O
the	O
nucleoprotein	B-protein
complex	I-protein
in	O
THP-1	B-cell_line
cells	I-cell_line
but	O
not	O
in	O
the	O
VD3-treated	B-cell_line
HL-60	I-cell_line
cells	I-cell_line
,	O
whereas	O
an	O
antibody	O
for	O
C/EBP	B-protein
beta	I-protein
blocked	O
the	O
formation	O
of	O
the	O
complex	O
with	O
the	O
nuclear	B-protein
factor	I-protein
of	O
the	O
HL-60	B-cell_line
cells	I-cell_line
but	O
not	O
with	O
that	O
of	O
THP-1	B-cell_line
cells	I-cell_line
.	O

An	O
anti-C/EBP	B-protein
delta	I-protein
antibody	I-protein
had	O
no	O
effect	O
on	O
the	O
complex	O
in	O
either	O
cell	O
.	O

Thus	O
,	O
it	O
was	O
concluded	O
that	O
C/EBP	B-protein
alpha	I-protein
and	I-protein
-beta	I-protein
were	O
able	O
to	O
bind	O
to	O
the	O
C/EBP	B-protein
motif	I-protein
,	O
and	O
that	O
C/EBP	B-protein
alpha	I-protein
bound	O
to	O
the	O
motif	O
in	O
THP-1	B-cell_line
cells	I-cell_line
and	O
C/EBP	B-protein
beta	I-protein
bound	O
to	O
that	O
in	O
the	O
VD3-treated	B-cell_line
HL-60	I-cell_line
cells	I-cell_line
.	O

Furthermore	O
,	O
to	O
examine	O
the	O
transcriptional	O
activity	O
of	O
the	O
C/EBP	B-protein
motif	I-protein
,	O
we	O
transfected	O
several	O
constructed	O
luciferase	B-DNA
reporter	I-DNA
DNAs	I-DNA
into	O
HL-60	B-cell_line
cells	I-cell_line
and	O
THP-1	B-cell_line
cells	I-cell_line
.	O

The	O
luciferase	O
activity	O
of	O
the	O
C/EBP	B-protein
motif	I-protein
in	O
HL-60	B-cell_line
cells	I-cell_line
was	O
increased	O
by	O
VD3	O
treatment	O
.	O

The	O
C/EBP	B-protein
motif	I-protein
in	O
the	O
MRP14	B-DNA
gene	I-DNA
was	O
confirmed	O
to	O
function	O
as	O
a	O
regulatory	O
region	O
in	O
VD3-treated	B-cell_line
HL-60	I-cell_line
cells	I-cell_line
and	I-cell_line
THP-1	I-cell_line
cells	I-cell_line
by	O
the	O
assay	O
.	O

Since	O
C/EBP	B-protein
beta	I-protein
was	O
also	O
detected	O
in	O
VD3-untreated	B-cell_line
HL-60	I-cell_line
cells	I-cell_line
by	O
immunoblotting	O
,	O
VD3	O
activated	O
C/EBP	B-protein
beta	I-protein
to	O
bind	O
to	O
the	O
motif	O
,	O
probably	O
through	O
post-translational	O
modification	O
.	O

-DOCSTART-	O

Patients	O
with	O
high-risk	O
myelodysplastic	O
syndrome	O
can	O
have	O
polyclonal	O
or	O
clonal	O
haemopoiesis	O
in	O
complete	O
haematological	O
remission	O
.	O

The	O
clonality	O
of	O
mature	B-cell_type
peripheral	I-cell_type
blood-derived	I-cell_type
myeloid	I-cell_type
and	I-cell_type
lymphoid	I-cell_type
cells	I-cell_type
and	O
bone	B-cell_type
marrow	I-cell_type
haemopoietic	I-cell_type
progenitors	I-cell_type
from	O
18	O
females	O
with	O
myelodysplasia	O
(	O
MDS	O
)	O
(	O
five	O
refractory	O
anaemia	O
,	O
RA	O
;	O
one	O
RA	O
with	O
ringed	B-cell_type
sideroblasts	I-cell_type
,	O
RARS	O
;	O
three	O
chronic	O
myelomonocytic	O
leukaemia	O
,	O
CMML	O
;	O
four	O
RA	O
with	O
excess	O
of	O
blasts	O
,	O
RAEB	O
;	O
five	O
RAEB	O
in	O
transformation	O
,	O
RAEB-t	O
)	O
was	O
studied	O
by	O
X-chromosome	B-DNA
inactivation	O
analysis	O
.	O

Using	O
the	O
human	O
androgen-receptor	O
(	O
HUMARA	O
)	O
assay	O
,	O
we	O
analysed	O
the	O
clonal	O
patterns	O
of	O
highly	B-cell_type
purified	I-cell_type
immature	I-cell_type
CD34+	I-cell_type
38-	I-cell_type
and	I-cell_type
committed	I-cell_type
CD34+	I-cell_type
38+	I-cell_type
marrow-derived	I-cell_type
progenitors	I-cell_type
,	O
and	O
CD16+	B-cell_type
14-	I-cell_type
granulocytes	I-cell_type
,	O
CD14+	B-cell_type
monocytes	I-cell_type
,	O
CD3+	B-cell_type
T	I-cell_type
and	I-cell_type
CD19+	I-cell_type
B	I-cell_type
lymphocytes	I-cell_type
from	O
peripheral	O
blood	O
.	O

In	O
high-risk	O
patients	O
(	O
RAEB	O
,	O
RAEB-t	O
)	O
,	O
clonality	O
analysis	O
was	O
performed	O
before	O
and	O
after	O
intensive	O
remission-induction	O
treatment	O
.	O

All	O
patients	O
,	O
except	O
one	O
with	O
RA	O
,	O
had	O
predominance	O
of	O
a	O
single	O
clone	O
in	O
their	O
granulocytes	B-cell_type
and	O
monocytes	B-cell_type
.	O

The	O
same	O
clonal	O
pattern	O
was	O
found	O
in	O
CD34+	B-cell_type
progenitor	I-cell_type
cells	I-cell_type
.	O

In	O
contrast	O
,	O
CD3+	B-cell_type
T	I-cell_type
lymphocytes	I-cell_type
were	O
polyclonal	O
or	O
oligoclonal	O
in	O
14/18	O
patients	O
.	O

X-chromosome	B-DNA
inactivation	O
patterns	O
of	O
CD19+	B-cell_type
B	I-cell_type
cells	I-cell_type
were	O
highly	O
concordant	O
with	O
CD3+	B-cell_type
T	I-cell_type
cells	I-cell_type
except	O
for	O
two	O
patients	O
(	O
one	O
RA	O
,	O
one	O
CMML	O
)	O
with	O
monoclonal	B-cell_type
B	I-cell_type
and	I-cell_type
polyclonal	I-cell_type
T	I-cell_type
lymphocytes	I-cell_type
,	O
therefore	O
suggesting	O
a	O
clonal	O
mutation	O
in	O
a	O
progenitor	B-cell_type
common	O
to	O
the	O
myeloid	B-cell_type
and	I-cell_type
B-lymphoid	I-cell_type
lineages	I-cell_type
or	O
the	O
coexistence	O
of	O
MDS	O
and	O
a	O
B-cell	O
disorder	O
in	O
these	O
particular	O
patients	O
.	O

After	O
high-dose	O
non-myeloablative	O
chemotherapy	O
,	O
polyclonal	O
haemopoiesis	O
was	O
reinstalled	O
in	O
the	O
mature	B-cell_type
myeloid	I-cell_type
cells	I-cell_type
and	O
immature	O
and	O
committed	B-cell_type
marrow	I-cell_type
progenitors	I-cell_type
in	O
three	O
of	O
four	O
patients	O
achieving	O
complete	O
haematological	O
remission	O
.	O

Therefore	O
we	O
conclude	O
that	O
most	O
haematological	O
remissions	O
in	O
MDS	O
are	O
associated	O
with	O
restoration	O
of	O
polyclonal	O
haemopoiesis	O
.	O

-DOCSTART-	O

Prominent	O
sex	O
steroid	O
metabolism	O
in	O
human	B-cell_type
lymphocytes	I-cell_type
.	O

Steroid	O
metabolism	O
was	O
investigated	O
in	O
cultured	B-cell_line
human	I-cell_line
B-lymphoblastoid	I-cell_line
cells	I-cell_line
(	O
B-LCL	B-cell_line
)	O
,	O
and	O
peripheral	B-cell_type
blood	I-cell_type
T	I-cell_type
and	I-cell_type
B	I-cell_type
cells	I-cell_type
.	O

Gene	O
expression	O
was	O
examined	O
by	O
reverse-transcription	O
polymerase	O
chain	O
reaction	O
amplification	O
(	O
RT-PCR	O
)	O
.	O

Appropriate	O
sized	O
transcripts	B-RNA
were	O
detected	O
in	O
both	O
cultured	O
and	O
fresh	O
peripheral	B-cell_type
lymphocytes	I-cell_type
for	O
CYP11A	B-protein
,	O
CYP17	B-protein
,	O
HSD11L	B-protein
(	O
11beta-hydroxysteroid	B-protein
dehydrogenase	I-protein
I	I-protein
)	O
,	O
HSD17B1	B-protein
(	O
17beta-hydroxysteroid	B-protein
dehydrogenase	I-protein
type	I-protein
I	I-protein
)	O
and	O
SRD5A1	B-protein
(	O
5alpha-reductase	B-protein
I	I-protein
)	O
.	O

B-LCL	O
,	O
but	O
not	O
T	B-cell_type
and	I-cell_type
B	I-cell_type
cells	I-cell_type
,	O
expressed	O
CYP11B	B-protein
.	O

There	O
was	O
minimal	O
expression	O
of	O
HSD3B1	B-protein
and	O
HSD3B2	B-protein
(	O
3beta-hydroxysteroid	B-protein
dehydrogenase	I-protein
I	I-protein
and	I-protein
II	I-protein
)	O
in	O
B-LCL	B-cell_line
and	O
T	B-cell_type
cells	I-cell_type
.	O

Transcripts	O
for	O
CYP19	B-protein
and	O
HSD11K	B-protein
were	O
not	O
detected	O
.	O

Corresponding	O
enzymatic	O
activity	O
was	O
detectable	O
only	O
for	O
17-hydroxysteroid	B-protein
dehydrogenase	I-protein
and	O
5alpha-reductase	B-protein
,	O
respectively	O
producing	O
testosterone	O
and	O
5alpha-dihydrotestosterone	O
.	O

Steroid	O
identities	O
were	O
confirmed	O
by	O
gas	O
chromatography/mass	O
spectrometry	O
(	O
GC/MS	O
)	O
.	O

One	O
metabolite	O
thought	O
to	O
be	O
deoxycorticosterone	O
was	O
identified	O
by	O
GC/MS	O
as	O
6alpha-hydroxypregnanolone	O
.	O

It	O
was	O
concluded	O
that	O
sex	O
hormone	O
metabolism	O
,	O
including	O
androgen	O
synthesis	O
,	O
occurs	O
in	O
lymphocytes	O
,	O
and	O
may	O
modulate	O
immune	O
response	O
.	O

-DOCSTART-	O

Human	B-cell_type
white	I-cell_type
blood	I-cell_type
cells	I-cell_type
and	O
hair	O
follicles	O
are	O
good	O
sources	O
of	O
mRNA	B-RNA
for	O
the	O
pterin	B-protein
carbinolamine	I-protein
dehydratase/dimerization	I-protein
cofactor	I-protein
of	O
HNF1	B-protein
for	O
mutation	O
detection	O
.	O

Pterin	B-protein
carbinolamine	I-protein
dehydratase/dimerization	I-protein
cofactor	I-protein
of	O
HNF1	B-protein
(	O
PCD/DCoH	B-protein
)	O
is	O
a	O
protein	O
that	O
has	O
a	O
dual	O
function	O
.	O

It	O
is	O
a	O
pterin	B-protein
4alpha-carbinolamine	I-protein
dehydratase	I-protein
that	O
is	O
involved	O
in	O
the	O
regeneration	O
of	O
the	O
cofactor	B-protein
tetrahydrobiopterin	I-protein
during	O
the	O
phenylalanine	O
hydroxylase-	O
catalyzed	O
hydroxylation	O
of	O
phenylalanine	O
.	O

In	O
addition	O
,	O
it	O
is	O
the	O
dimerization	O
cofactor	O
of	O
HNF1	B-protein
that	O
is	O
able	O
to	O
activate	O
the	O
transcriptional	O
activity	O
of	O
HNF1	B-protein
.	O

Deficiencies	O
in	O
the	O
gene	O
for	O
this	O
dual	B-protein
functional	I-protein
protein	I-protein
result	O
in	O
hyperphenylalaninemia	O
.	O

Here	O
we	O
report	O
for	O
the	O
first	O
time	O
that	O
the	O
PCD/DCoH	B-RNA
mRNA	I-RNA
is	O
present	O
in	O
human	B-cell_type
white	I-cell_type
blood	I-cell_type
cells	I-cell_type
and	O
hair	O
follicles	O
.	O

Taking	O
advantage	O
of	O
this	O
finding	O
,	O
a	O
sensitive	O
,	O
rapid	O
and	O
convenient	O
method	O
for	O
screening	O
mutations	O
occurring	O
in	O
the	O
coding	O
region	O
of	O
this	O
gene	O
has	O
been	O
described	O
.	O

Copyright	O
1998	O
Academic	O
Press	O
.	O

-DOCSTART-	O

Biochemical	O
characterization	O
of	O
the	O
NF-Y	B-protein
transcription	I-protein
factor	I-protein
complex	I-protein
during	O
B	O
lymphocyte	O
development	O
.	O

The	O
transcription	B-protein
factor	I-protein
,	O
NF-Y	B-protein
,	O
plays	O
a	O
critical	O
role	O
in	O
tissue-specific	O
major	B-DNA
histocompatibility	I-DNA
complex	I-DNA
class	I-DNA
II	I-DNA
gene	I-DNA
transcription	O
.	O

In	O
this	O
report	O
the	O
biochemical	O
properties	O
of	O
the	O
heterotrimeric	B-protein
NF-Y	I-protein
complex	I-protein
have	O
been	O
characterized	O
during	O
stage-specific	O
B-cell	O
development	O
,	O
and	O
in	O
several	O
class	B-cell_line
II-	I-cell_line
mutant	I-cell_line
B-cell	I-cell_line
lines	I-cell_line
,	O
which	O
represent	O
distinct	O
bare	O
lymphocyte	O
syndrome	O
class	O
II	O
genetic	O
complementation	O
groups	O
.	O

The	O
NF-Y	B-protein
complex	I-protein
derived	O
from	O
class	B-cell_type
II+	I-cell_type
mature	I-cell_type
B-cells	I-cell_type
bound	O
with	O
high	O
affinity	O
to	O
anion	B-protein
exchangers	I-protein
,	O
and	O
eluted	O
as	O
an	O
intact	O
trimeric	B-protein
complex	I-protein
,	O
whereas	O
,	O
NF-Y	B-protein
derived	O
from	O
class	B-cell_type
II-	I-cell_type
plasma	I-cell_type
B-cells	I-cell_type
,	O
and	O
from	O
bare	B-cell_line
lymphocyte	I-cell_line
syndrome	I-cell_line
group	I-cell_line
II	I-cell_line
cell	I-cell_line
lines	I-cell_line
,	O
RJ2.2.5	B-cell_line
and	O
RM3	B-cell_line
,	O
dissociated	O
into	O
discrete	O
NF-YA	B-protein
and	O
NF-YB	B-protein
:	I-protein
C	I-protein
subunit	B-protein
fractions	I-protein
.	O

Recombination	O
of	O
the	O
MPC11	B-protein
plasma	I-protein
B-cell	I-protein
derived	I-protein
NF-Y	I-protein
A	I-protein
:	I-protein
B	I-protein
:	I-protein
C	I-protein
complex	I-protein
with	O
the	O
low	O
molecular	O
mass	O
protein	O
fraction	O
,	O
NF-Y-associated	B-protein
factors	I-protein
(	O
YAFs	B-protein
)	O
,	O
derived	O
from	O
mature	O
A20	O
B-cell	O
nuclei	O
,	O
conferred	O
high	O
affinity	O
anion	O
exchange	O
binding	O
to	O
NF-Y	B-protein
as	O
an	O
intact	B-protein
trimeric	I-protein
complex	I-protein
.	O

Recombination	O
of	O
the	O
native	B-protein
NF-YA	I-protein
:	I-protein
B	I-protein
:	I-protein
C	I-protein
complex	I-protein
with	O
the	O
transcriptional	B-protein
cofactor	I-protein
,	O
PC4	B-protein
,	O
likewise	O
conferred	O
high	B-protein
affinity	I-protein
NF-Y	I-protein
binding	O
to	O
anion	B-protein
exchangers	I-protein
,	O
and	O
stabilized	O
NF-Y	B-protein
interaction	O
with	O
CCAAT-box	B-DNA
DNA	I-DNA
motifs	I-DNA
in	O
vitro	O
.	O

Interaction	O
between	O
PC4	B-protein
and	O
NF-Y	B-protein
was	O
mapped	O
to	O
the	O
C-terminal	B-protein
region	I-protein
of	O
PC4	B-protein
,	O
and	O
the	O
subunit	B-protein
interaction	I-protein
subdomain	I-protein
of	O
the	O
highly	B-protein
conserved	I-protein
DNA	I-protein
binding-subunit	I-protein
interaction	I-protein
domain	I-protein
(	O
DBD	B-protein
)	O
of	O
NF-YA	B-protein
.	O

These	O
results	O
suggest	O
that	O
in	O
class	B-cell_type
II+	I-cell_type
mature	I-cell_type
B-cells	I-cell_type
NF-Y	B-protein
is	O
associated	O
with	O
the	O
protein	O
cofactor	O
,	O
PC4	B-protein
,	O
which	O
may	O
play	O
an	O
important	O
role	O
in	O
NF-Y-mediated	O
transcriptional	O
control	O
of	O
class	B-DNA
II	I-DNA
genes	I-DNA
.	O

-DOCSTART-	O

Arrest	O
of	O
B	O
lymphocyte	O
terminal	O
differentiation	O
by	O
CD40	O
signaling	O
:	O
mechanism	O
for	O
lack	O
of	O
antibody-secreting	B-cell_type
cells	I-cell_type
in	O
germinal	O
centers	O
.	O

Despite	O
extensive	O
research	O
,	O
the	O
role	O
of	O
CD40	B-protein
signaling	O
in	O
B	O
cell	O
terminal	O
differentiation	O
remains	O
controversial	O
.	O

Here	O
we	O
show	O
that	O
CD40	B-protein
engagement	O
arrests	O
B	O
cell	O
differentiation	O
prior	O
to	O
plasma	O
cell	O
formation	O
.	O

This	O
arrest	O
is	O
manifested	O
at	O
a	O
molecular	O
level	O
as	O
a	O
reduction	O
in	O
mRNA	O
levels	O
of	O
secretory	O
immunoglobulin	O
gene	O
products	O
such	O
as	O
mu	O
(	O
s	O
)	O
and	O
J	O
chain	O
as	O
well	O
as	O
the	O
loss	O
of	O
the	O
transcriptional	B-protein
regulator	I-protein
BLIMP-1	B-protein
.	O

Furthermore	O
,	O
the	O
inhibition	O
of	O
B	O
cell	O
differentiation	O
by	O
CD40	B-protein
engagement	O
could	O
not	O
be	O
overcome	O
by	O
either	O
mitogens	B-protein
or	O
cytokines	B-protein
,	O
but	O
could	O
be	O
reversed	O
by	O
antibodies	O
that	O
interfere	O
with	O
the	O
CD40	B-protein
/	O
gp39	B-protein
interaction	O
.	O

These	O
data	O
suggest	O
that	O
secretory	O
immunoglobulin	O
is	O
not	O
produced	O
by	O
B	B-cell_type
cells	I-cell_type
that	O
are	O
actively	O
engaged	O
by	O
gp39-expressing	B-cell_type
T	I-cell_type
cells	I-cell_type
.	O

-DOCSTART-	O

A	O
positively	O
charged	O
alpha-lipoic	O
acid	O
analogue	O
with	O
increased	O
cellular	O
uptake	O
and	O
more	O
potent	O
immunomodulatory	O
activity	O
.	O

alpha-Lipoic	O
acid	O
(	O
LA	O
)	O
is	O
taken	O
up	O
by	O
cells	O
and	O
reduced	O
to	O
its	O
potent	O
dithiol	O
form	O
,	O
dihydrolipoate	O
(	O
DHLA	O
)	O
,	O
much	O
of	O
which	O
is	O
rapidly	O
effluxed	O
out	O
from	O
cells	O
.	O

To	O
improve	O
retention	O
in	O
cells	O
,	O
the	O
LA	O
molecule	O
was	O
modified	O
to	O
confer	O
a	O
positive	O
charge	O
at	O
physiological	O
pH	O
.	O

N	O
,	O
N-dimethyl	O
,	O
N'-2-amidoethyl-lipoate	O
was	O
synthesized	O
.	O

The	O
protonated	O
form	O
of	O
the	O
new	O
molecule	O
is	O
referred	O
to	O
as	O
LA-Plus	O
.	O

The	O
uptake	O
of	O
LA-Plus	O
by	O
human	B-cell_type
Wurzburg	I-cell_type
T	I-cell_type
cells	I-cell_type
was	O
higher	O
compared	O
to	O
that	O
of	O
LA	O
.	O

Several-fold	O
higher	O
amounts	O
of	O
DHLA-Plus	O
,	O
the	O
corresponding	O
reduced	O
form	O
of	O
LA-Plus	O
,	O
were	O
detected	O
in	O
LA-Plus	O
treated	O
cells	O
compared	O
to	O
the	O
amount	O
of	O
DHLA	O
found	O
in	O
cells	O
treated	O
with	O
LA	O
.	O

At	O
100	O
microM	O
,	O
LA	O
did	O
not	O
but	O
LA-Plus	O
inhibited	O
H2O2	O
induced	O
NF-kappaB	B-protein
activation	O
and	O
NF-kappaB	B-protein
directed	O
IL-2	B-protein
receptor	I-protein
expression	O
.	O

Both	O
LA	O
and	O
LA-Plus	O
synergised	O
with	O
selenium	O
in	O
inhibiting	O
H2O2	O
induced	O
NF-kappaB	B-protein
activation	O
.	O

At	O
150	O
microM	O
LA-Plus	O
,	O
but	O
not	O
LA	O
,	O
inhibited	O
TNFalpha	B-protein
induced	O
NF-kappaB	B-protein
activation	O
.	O

At	O
5	O
microM	O
LA-Plus	O
,	O
but	O
not	O
LA	O
,	O
protected	O
against	O
both	O
spontaneous	O
and	O
etoposide	O
induced	O
apoptosis	O
in	O
rat	B-cell_type
thymocytes	I-cell_type
.	O

LA-Plus	O
is	O
thus	O
an	O
improved	O
form	O
of	O
LA	O
with	O
increased	O
therapeutic	O
potential	O
.	O

Copyright	O
1998	O
Academic	O
Press	O
.	O

-DOCSTART-	O

Carrier	O
identification	O
in	O
X-linked	O
immunodeficiency	O
diseases	O
.	O

OBJECTIVE	O
:	O
Carrier	O
identification	O
in	O
X-linked	O
immunodeficiency	O
disorders	O
can	O
be	O
based	O
on	O
the	O
demonstration	O
of	O
non-random	O
X	O
inactivation	O
(	O
NRXI	O
)	O
in	O
affected	B-cell_type
blood	I-cell_type
cell	I-cell_type
lineages	I-cell_type
when	O
growth	O
is	O
impaired	O
in	O
cells	O
expressing	O
the	O
abnormal	B-DNA
gene	I-DNA
.	O

We	O
examined	O
the	O
utility	O
of	O
seeking	O
evidence	O
of	O
NRXI	O
to	O
test	O
the	O
carrier	O
status	O
of	O
women	O
in	O
families	O
affected	O
by	O
X-linked	O
severe	O
combined	O
immunodeficiency	O
(	O
XSCID	O
)	O
and	O
X-linked	O
hypogammaglobulinaemia	O
(	O
XLH	O
)	O
,	O
to	O
identify	O
as	O
carriers	O
the	O
mothers	O
of	O
boys	O
with	O
SCID	O
or	O
hypogammaglobulinaemia	O
whose	O
phenotype	O
suggested	O
X-linkage	O
and	O
to	O
infer	O
X-linkage	O
in	O
boys	O
with	O
SCID	O
or	O
hypogammaglobulinaemia	O
whose	O
disease	O
was	O
not	O
clearly	O
X-linked	O
on	O
the	O
basis	O
either	O
of	O
family	O
history	O
or	O
clinical	O
and	O
immunological	O
characteristics	O
.	O

METHODOLOGY	O
:	O
A	O
polymerase	O
chain	O
reaction-based	O
method	O
was	O
used	O
to	O
amplify	O
a	O
polymorphic	O
CAG	B-DNA
repeat	I-DNA
in	O
the	O
first	B-DNA
exon	I-DNA
of	O
the	O
androgen	B-DNA
receptor	I-DNA
gene	I-DNA
after	O
selective	O
digestion	O
of	O
the	O
active	B-DNA
X	I-DNA
chromosome	I-DNA
with	O
a	O
methylation-sensitive	B-protein
enzyme	I-protein
,	O
HpaII	B-protein
to	O
distinguish	O
between	O
the	O
paternal	B-DNA
and	I-DNA
maternal	I-DNA
alleles	I-DNA
and	O
to	O
identify	O
their	O
methylation	O
status	O
.	O

RESULTS	O
:	O
Heterozygosity	O
was	O
found	O
in	O
24	O
of	O
31	O
female	O
subjects	O
(	O
77	O
%	O
)	O
.	O

As	O
anticipated	O
,	O
NRXI	O
could	O
be	O
demonstrated	O
in	O
all	O
lymphoid	B-cell_type
cells	I-cell_type
studied	O
from	O
obligate	O
carriers	O
of	O
XSCID	O
and	O
an	O
obligate	O
carrier	O
of	O
XLH	O
but	O
not	O
on	O
a	O
carrier	O
of	O
X-linked	O
immunodeficiency	O
with	O
hyper-IgM	B-protein
.	O

The	O
finding	O
of	O
NRXI	O
in	O
the	O
mother	O
of	O
a	O
boy	O
with	O
a	O
SCID	O
variant	O
showed	O
her	O
to	O
be	O
a	O
carrier	O
of	O
XSCID	O
and	O
establishes	O
that	O
her	O
son	O
has	O
XSCID	O
,	O
not	O
otherwise	O
evident	O
from	O
available	O
data	O
.	O

CONCLUSIONS	O
:	O
This	O
PCR	O
assay	O
provides	O
a	O
rapid	O
method	O
for	O
carrier	O
detection	O
of	O
X-linked	O
immunodeficiencies	O
,	O
and	O
has	O
allowed	O
us	O
to	O
expand	O
the	O
phenotype	O
of	O
XSCID	O

-DOCSTART-	O

Leukocyte-endothelial	O
interaction	O
is	O
augmented	O
by	O
high	O
glucose	O
concentrations	O
and	O
hyperglycemia	O
in	O
a	O
NF-kB-dependent	O
fashion	O
.	O

We	O
addressed	O
the	O
role	O
of	O
hyperglycemia	O
in	O
leukocyte-endothelium	O
interaction	O
under	O
flow	O
conditions	O
by	O
exposing	O
human	B-cell_type
umbilical	I-cell_type
vein	I-cell_type
endothelial	I-cell_type
cells	I-cell_type
for	O
24	O
h	O
to	O
normal	O
(	O
5	O
mM	O
)	O
,	O
high	O
concentration	O
of	O
glucose	O
(	O
30	O
mM	O
)	O
,	O
advanced	O
glycosylation	O
end	O
product-albumin	B-protein
(	O
100	O
microg/ml	O
)	O
,	O
or	O
hyperglycemic	O
(	O
174-316	O
mg/dl	O
)	O
sera	O
from	O
patients	O
with	O
diabetes	O
and	O
abnormal	O
hemoglobin	O
A1c	O
(	O
8.1+/-1.4	O
%	O
)	O
.	O

At	O
the	O
end	O
of	O
incubation	O
endothelial	B-cell_type
cells	I-cell_type
were	O
perfused	O
with	O
total	O
leukocyte	O
suspension	O
in	O
a	O
parallel	O
plate	O
flow	O
chamber	O
under	O
laminar	O
flow	O
(	O
1.5	O
dyn/cm2	O
)	O
.	O

Rolling	O
and	O
adherent	B-cell_type
cells	I-cell_type
were	O
evaluated	O
by	O
digital	O
image	O
processing	O
.	O

Results	O
showed	O
that	O
30	O
mM	O
glucose	O
significantly	O
(	O
P	O
<	O
0.01	O
)	O
increased	O
the	O
number	O
of	O
adherent	B-cell_type
leukocytes	I-cell_type
to	O
endothelial	B-cell_type
cells	I-cell_type
in	O
respect	O
to	O
control	O
(	O
5	O
mM	O
glucose	O
;	O
151+/-19	O
versus	O
33+/-8	O
cells/mm2	O
)	O
.	O

A	O
similar	O
response	O
was	O
induced	O
by	O
endothelial	O
stimulation	O
with	O
IL-1beta	B-protein
,	O
here	O
used	O
as	O
positive	O
control	O
(	O
195+/-20	O
cells/mm2	O
)	O
.	O

The	O
number	O
of	O
rolling	B-cell_type
cells	I-cell_type
on	O
endothelial	O
surface	O
was	O
not	O
affected	O
by	O
high	O
glucose	O
level	O
.	O

Stable	O
adhesion	O
of	O
leukocytes	B-cell_type
to	O
glucose-treated	O
as	O
well	O
as	O
to	O
IL-1beta-stimulated	B-cell_type
endothelial	I-cell_type
cells	I-cell_type
was	O
preceded	O
by	O
short	O
interaction	O
of	O
leukocytes	B-cell_type
with	O
the	O
endothelial	O
surface	O
.	O

The	O
distance	O
travelled	O
by	O
leukocytes	B-cell_type
before	O
arrest	O
on	O
30	O
mM	O
glucose	O
,	O
or	O
on	O
IL-1beta-treated	B-cell_type
endothelial	I-cell_type
cells	I-cell_type
,	O
was	O
significantly	O
(	O
P	O
<	O
0.01	O
)	O
higher	O
than	O
that	O
observed	O
for	O
leukocytes	B-cell_type
adhering	O
on	O
control	O
endothelium	O
(	O
30	O
mM	O
glucose	O
:	O
76.7+/-3.5	O
;	O
IL1beta	O
:	O
69.7+/-4	O
versus	O
5	O
mM	O
glucose	O
:	O
21.5+/-5	O
microm	O
)	O
.	O

Functional	O
blocking	O
of	O
E-selectin	B-protein
,	O
intercellular	B-protein
cell	I-protein
adhesion	I-protein
molecule-1	I-protein
,	O
and	O
vascular	B-protein
cell	I-protein
adhesion	I-protein
molecule-1	I-protein
on	O
endothelial	B-cell_type
cells	I-cell_type
with	O
the	O
corresponding	O
mouse	B-protein
mAb	I-protein
significantly	O
inhibited	O
glucose-induced	O
increase	O
in	O
leukocyte	O
adhesion	O
(	O
67+/-16	O
,	O
83+/-12	O
,	O
62+/-8	O
versus	O
144+/-21	O
cells/	O
mm2	O
)	O
.	O

Confocal	O
fluorescence	O
microscopy	O
studies	O
showed	O
that	O
30	O
mM	O
glucose	O
induced	O
an	O
increase	O
in	O
endothelial	O
surface	O
expression	O
of	O
E-selectin	B-protein
,	O
intercellular	B-protein
cell	I-protein
adhesion	I-protein
molecule-1	I-protein
,	O
and	O
vascular	B-protein
cell	I-protein
adhesion	I-protein
molecule-1	I-protein
.	O

Electrophoretic	O
mobility	O
shift	O
assay	O
of	O
nuclear	O
extracts	O
of	O
human	B-cell_type
umbilical	I-cell_type
vein	I-cell_type
endothelial	I-cell_type
cells	I-cell_type
(	O
HUVEC	O
)	O
exposed	O
for	O
1	O
h	O
to	O
30	O
mM	O
glucose	O
revealed	O
an	O
intense	O
NF-kB	B-protein
activation	O
.	O

Treatment	O
of	O
HUVEC	O
exposed	O
to	O
high	O
glucose	O
with	O
the	O
NF-kB	B-protein
inhibitors	I-protein
pyrrolidinedithiocarbamate	O
(	O
100	O
microM	O
)	O
and	O
tosyl-phe-chloromethylketone	O
(	O
25	O
microM	O
)	O
significantly	O
reduced	O
(	O
P	O
<	O
0.05	O
)	O
leukocyte	O
adhesion	O
in	O
respect	O
to	O
HUVEC	O
treated	O
with	O
glucose	O
alone	O
.	O

A	O
significant	O
(	O
P	O
<	O
0.01	O
)	O
inhibitory	O
effect	O
on	O
glucose-induced	O
leukocyte	O
adhesion	O
was	O
observed	O
after	O
blocking	O
protein	B-protein
kinase	I-protein
C	I-protein
activity	O
with	O
staurosporine	O
(	O
5	O
nM	O
)	O
.	O

When	O
HUVEC	O
were	O
treated	O
with	O
specific	O
antisense	O
oligodesoxynucleotides	O
against	O
PKCalpha	B-protein
and	I-protein
PKCepsilon	I-protein
isoforms	I-protein
before	O
the	O
addition	O
of	O
30	O
mM	O
glucose	O
,	O
a	O
significant	O
(	O
P	O
<	O
0.05	O
)	O
reduction	O
in	O
the	O
adhesion	O
was	O
also	O
seen	O
.	O

Advanced	O
glycosylation	O
end	O
product-albumin	B-protein
significantly	O
increased	O
the	O
number	O
of	O
adhering	B-cell_type
leukocytes	I-cell_type
in	O
respect	O
to	O
native	B-protein
albumin	I-protein
used	O
as	O
control	O
(	O
110+/-16	O
versus	O
66+/-7	O
,	O
P	O
<	O
0.01	O
)	O
.	O

Sera	O
from	O
diabetic	O
patients	O
significantly	O
(	O
P	O
<	O
0.01	O
)	O
enhanced	O
leukocyte	O
adhesion	O
as	O
compared	O
with	O
controls	O
,	O
despite	O
normal	O
levels	O
of	O
IL-1beta	B-protein
and	O
TNFalpha	B-protein
in	O
these	O
sera	O
.	O

These	O
data	O
indicate	O
that	O
high	O
glucose	O
concentration	O
and	O
hyperglycemia	O
promote	O
leukocyte	O
adhesion	O
to	O
the	O
endothelium	O
through	O
upregulation	O
of	O
cell	O
surface	O
expression	O
of	O
adhesive	O
proteins	O
,	O
possibly	O
depending	O
on	O
NF-kB	B-protein
activation	O
.	O

-DOCSTART-	O

Ikaros	B-protein
in	O
hemopoietic	O
lineage	O
determination	O
and	O
homeostasis	O
.	O

Studies	O
on	O
the	O
molecular	O
mechanisms	O
that	O
control	O
hemopoietic	O
differentiation	O
have	O
focused	O
on	O
signaling	O
cascades	O
and	O
nuclear	O
effectors	O
that	O
drive	O
this	O
complex	O
developmental	O
system	O
in	O
a	O
regulated	O
fashion	O
.	O

Here	O
we	O
review	O
the	O
role	O
of	O
Ikaros	B-protein
,	O
the	O
founding	O
member	O
of	O
a	O
unique	O
family	O
of	O
zinc	B-protein
finger	I-protein
transcription	I-protein
factors	I-protein
in	O
this	O
developmental	O
process	O
.	O

Studies	O
on	O
an	O
Ikaros	B-protein
null	O
mutation	O
have	O
revealed	O
an	O
essential	O
role	O
for	O
this	O
factor	O
in	O
lymphoid	O
cell	O
fate	O
determination	O
and	O
at	O
subsequent	O
branch	O
points	O
of	O
the	O
T	O
cell	O
differentiation	O
pathway	O
.	O

Differences	O
in	O
the	O
phenotypes	O
of	O
a	O
null	O
and	O
a	O
dominant	B-DNA
negative	I-DNA
(	I-DNA
DN	I-DNA
)	I-DNA
Ikaros	I-DNA
mutation	I-DNA
provide	O
insight	O
into	O
a	O
regulatory	O
network	O
through	O
which	O
Ikaros	B-protein
proteins	I-protein
exert	O
their	O
effects	O
in	O
development	O
.	O

In	O
addition	O
a	O
comparative	O
analysis	O
of	O
the	O
hemopoietic	B-cell_type
stem	I-cell_type
cell	I-cell_type
and	O
precursor	O
compartment	O
resulting	O
from	O
the	O
two	O
Ikaros	B-DNA
mutations	I-DNA
reveals	O
a	O
profound	O
yet	O
not	O
absolute	O
requirement	O
for	O
Ikaros	B-protein

-DOCSTART-	O

Induction	O
of	O
early	B-protein
B	I-protein
cell	I-protein
factor	I-protein
(	O
EBF	B-protein
)	O
and	O
multiple	B-DNA
B	I-DNA
lineage	I-DNA
genes	I-DNA
by	O
the	O
basic	B-protein
helix-loop-helix	I-protein
transcription	I-protein
factor	I-protein
E12	B-protein
.	O

The	O
transcription	B-protein
factors	I-protein
encoded	O
by	O
the	O
E2A	B-protein
and	O
early	B-protein
B	I-protein
cell	I-protein
factor	I-protein
(	O
EBF	B-protein
)	O
genes	O
are	O
required	O
for	O
the	O
proper	O
development	O
of	O
B	B-cell_type
lymphocytes	I-cell_type
.	O

However	O
,	O
the	O
absence	O
of	O
B	B-cell_type
lineage	I-cell_type
cells	I-cell_type
in	O
E2A-	O
and	O
EBF-deficient	O
mice	O
has	O
made	O
it	O
difficult	O
to	O
determine	O
the	O
function	O
or	O
relationship	O
between	O
these	O
proteins	O
.	O

We	O
report	O
the	O
identification	O
of	O
a	O
novel	O
model	O
system	O
in	O
which	O
the	O
role	O
of	O
E2A	B-protein
and	O
EBF	B-protein
in	O
the	O
regulation	O
of	O
multiple	O
B	O
lineage	O
traits	O
can	O
be	O
studied	O
.	O

We	O
found	O
that	O
the	O
conversion	O
of	O
70Z/3	B-cell_line
pre-B	I-cell_line
lymphocytes	I-cell_line
to	O
cells	O
with	O
a	O
macrophage-like	O
phenotype	O
is	O
associated	O
with	O
the	O
loss	O
of	O
E2A	B-protein
and	O
EBF	B-protein
.	O

Moreover	O
,	O
we	O
show	O
that	O
ectopic	O
expression	O
of	O
the	O
E2A	B-protein
protein	O
E12	B-protein
in	O
this	O
macrophage	B-cell_line
line	I-cell_line
results	O
in	O
the	O
induction	O
of	O
many	B-DNA
B	I-DNA
lineage	I-DNA
genes	I-DNA
,	O
including	O
EBF	B-protein
,	O
IL7Ralpha	B-protein
,	O
lambda5	B-protein
,	O
and	O
Rag-1	O
,	O
and	O
the	O
ability	O
to	O
induce	O
kappa	B-protein
light	I-protein
chain	I-protein
in	O
response	O
to	O
mitogen	O
.	O

Activation	O
of	O
EBF	B-protein
may	O
be	O
one	O
of	O
the	O
critical	O
functions	O
of	O
E12	B-protein
in	O
regulating	O
the	O
B	O
lineage	O
phenotype	O
since	O
expression	O
of	O
EBF	B-protein
alone	O
leads	O
to	O
the	O
activation	O
of	O
a	O
subset	O
of	O
E12	B-protein
-inducible	O
traits	O
.	O

Our	O
data	O
demonstrate	O
that	O
,	O
in	O
the	O
context	O
of	O
this	O
macrophage	B-cell_line
line	I-cell_line
,	O
E12	B-protein
induces	O
expression	O
of	O
EBF	B-protein
and	O
together	O
these	O
transcription	B-protein
factors	I-protein
coordinately	O
regulate	O
numerous	O
B	B-DNA
lineage-associated	I-DNA
genes	I-DNA
.	O

-DOCSTART-	O

Activation-induced	O
down-regulation	O
of	O
retinoid	B-protein
receptor	I-protein
RXRalpha	I-protein
expression	O
in	O
human	B-cell_type
T	I-cell_type
lymphocytes	I-cell_type
.	O

Role	O
of	O
cell	O
cycle	O
regulation	O
.	O

A	O
5.4-kilobase	B-RNA
mRNA	I-RNA
,	O
the	O
expression	O
of	O
which	O
is	O
down-regulated	O
after	O
treatment	O
of	O
human	B-cell_type
peripheral	I-cell_type
blood	I-cell_type
mononuclear	I-cell_type
cells	I-cell_type
(	O
PBMCs	B-cell_type
)	O
with	O
various	O
T	O
cell-activating	O
agents	O
,	O
was	O
isolated	O
using	O
an	O
mRNA	O
differential	O
display	O
method	O
.	O

Nucleotide	O
sequence	O
analysis	O
identified	O
the	O
5	B-RNA
'	I-RNA
end	I-RNA
of	O
this	O
RNA	B-RNA
as	O
human	B-RNA
retinoid	I-RNA
receptor	I-RNA
RXRalpha	I-RNA
mRNA	I-RNA
.	O

Here	O
,	O
we	O
report	O
the	O
nucleotide	O
sequence	O
of	O
3.6	B-DNA
kilobases	I-DNA
of	O
this	O
RNA	O
,	O
which	O
represents	O
the	O
3	B-RNA
'	I-RNA
end	I-RNA
of	O
RXRalpha	B-RNA
mRNA	I-RNA
,	O
the	O
sequence	O
of	O
which	O
has	O
not	O
been	O
previously	O
described	O
.	O

Activated	O
PBMCs	B-cell_type
also	O
expressed	O
lower	O
levels	O
of	O
RXRalpha	B-protein
protein	O
,	O
and	O
a	O
DNA	O
binding	O
assay	O
showed	O
that	O
the	O
activation-induced	O
loss	O
of	O
RXRalpha	B-RNA
mRNA	I-RNA
and	O
protein	O
expression	O
correlated	O
with	O
the	O
loss	O
of	O
DNA	O
binding	O
activity	O
of	O
this	O
protein	O
.	O

We	O
present	O
evidence	O
that	O
the	O
transition	O
from	O
G0/G1	O
to	O
S	O
phase	O
of	O
the	O
cell	O
cycle	O
results	O
in	O
the	O
down-regulation	O
of	O
RXRalpha	B-protein
expression	O
and	O
that	O
cell	O
cycle	O
inhibitors	O
,	O
which	O
block	O
the	O
cells	O
in	O
G1	O
phase	O
,	O
prevent	O
this	O
down-regulation	O
.	O

The	O
decrease	O
in	O
the	O
levels	O
of	O
RXRalpha	B-RNA
mRNA	I-RNA
was	O
found	O
to	O
be	O
regulated	O
at	O
the	O
post-transcriptional	O
level	O
and	O
involved	O
new	O
protein	O
synthesis	O
.	O

These	O
observations	O
indicate	O
that	O
the	O
levels	O
of	O
RXRalpha	B-protein
expression	O
in	O
T	O
lymphocytes	O
are	O
coupled	O
to	O
cell	O
cycle	O
progression	O
,	O
and	O
there	O
is	O
tight	O
regulatory	O
control	O
of	O
RXRalpha	B-protein
expression	O
during	O
the	O
transition	O
from	O
G0/G1	O
to	O
S	O
phase	O
of	O
the	O
cell	O
cycle	O
.	O

-DOCSTART-	O

Epidemiology	O
and	O
pathogenesis	O
of	O
AIDS-related	O
lymphomas	O
.	O

Among	O
patients	O
with	O
congenital	O
and	O
acquired	O
immunodeficiencies	O
,	O
non-Hodgkin	O
's	O
lymphoma	O
(	O
NHLs	O
)	O
are	O
the	O
most	O
common	O
tumors	O
of	O
the	O
immune	O
system	O
.	O

In	O
the	O
setting	O
of	O
human	O
immunodeficiency	O
virus	O
(	O
HIV	O
)	O
infection	O
,	O
as	O
many	O
as	O
10	O
%	O
to	O
20	O
%	O
of	O
people	O
ultimately	O
developed	O
NHLs	O
.	O

These	O
tumors	O
are	O
clinically	O
aggressive	O
,	O
frequently	O
involve	O
extranodal	O
sites	O
,	O
and	O
often	O
exhibit	O
unique	O
features	O
that	O
distinguish	O
them	O
from	O
NHL	O
arising	O
in	O
individuals	O
with	O
other	O
forms	O
of	O
immunosuppression	O
.	O

Important	O
in	O
the	O
development	O
of	O
HIV-associated	O
NHL	O
are	O
cytokines	B-protein
and	O
other	O
factors	O
that	O
induce	O
B-cell	O
proliferation	O
and	O
increase	O
the	O
likelihood	O
of	O
mutations	O
of	O
c-myc	B-DNA
,	O
bcl-6	B-DNA
,	O
and	O
other	O
tumor-suppressor	B-DNA
genes	I-DNA
with	O
carcinogenic	O
potential	O
.	O

Specific	O
forms	O
of	O
HIV-associated	O
NHL	O
are	O
linked	O
to	O
expression	O
of	O
Epstein-Barr	B-protein
virus	I-protein
(	I-protein
EBV	I-protein
)	I-protein
-latent	I-protein
proteins	I-protein
;	O
the	O
newly	O
described	O
DNA	O
virus	O
,	O
Karposi	O
's	O
sarcoma-associated	O
herpesvirus/human	O
herpesvirus-8	O
(	O
KSHV/HHV-8	O
)	O
;	O
and	O
perhaps	O
HIV	O
.	O

Elucidation	O
of	O
the	O
factors	O
that	O
contribute	O
to	O
the	O
high	O
incidence	O
of	O
NHL	O
among	O
patients	O
infected	O
with	O
HIV	O
provides	O
insights	O
into	O
important	O
elements	O
of	O
lymphomagenesis	O
.	O

-DOCSTART-	O

Matrix	B-protein
metalloproteinase	I-protein
expression	O
in	O
human	O
breast	O
cancer	O
:	O
an	O
immunohistochemical	O
study	O
including	O
correlation	O
with	O
cathepsin	B-protein
D	I-protein
,	O
type	B-protein
IV	I-protein
collagen	I-protein
,	O
laminin	B-protein
,	O
fibronectin	B-protein
,	O
EGFR	B-protein
,	O
c-erbB-2	B-protein
oncoprotein	I-protein
,	O
p53	B-protein
,	O
steroid	O
receptors	O
status	O
and	O
proliferative	O
indices	O
.	O

Matrix	B-protein
metalloproteinase	I-protein
s	O
(	O
MMPs	B-protein
)	O
are	O
a	O
group	O
of	O
enzymes	O
thought	O
to	O
be	O
responsible	O
for	O
both	O
normal	O
connective	O
tissue	O
matrix	O
remodelling	O
and	O
accelerated	O
breakdown	O
associated	O
with	O
tumour	O
development	O
.	O

The	O
current	O
study	O
aimed	O
to	O
investigate	O
the	O
immunohistochemical	O
expression	O
of	O
matrix	B-protein
metalloproteinase	I-protein
3	I-protein
(	O
MMP-3	B-protein
,	O
stromelysin-1	B-protein
)	O
in	O
correlation	O
with	O
the	O
expression	O
of	O
Basement	B-protein
Membrane	I-protein
(	I-protein
BM	I-protein
)	I-protein
antigen	I-protein
(	O
type	B-protein
IV	I-protein
collagen	I-protein
,	O
laminin	B-protein
)	O
,	O
fibronectin	B-protein
,	O
cathepsin	B-protein
D	I-protein
,	O
p53	B-protein
,	O
c-erbB-2	B-protein
,	O
proliferative	O
activity	O
(	O
Ki-67	B-protein
,	O
PCNA	B-protein
)	O
,	O
steroid	O
receptor	O
content	O
as	O
well	O
as	O
to	O
the	O
other	O
conventional	O
clinicopathological	O
parameters	O
in	O
breast	O
cancer	O
.	O

This	O
study	O
was	O
performed	O
on	O
a	O
series	O
of	O
frozen	O
and	O
paraffin	O
sections	O
from	O
84	O
breast	O
cancer	O
specimens	O
by	O
immunohistochemistry	O
using	O
the	O
monoclonal	B-protein
antibody	I-protein
MMP-3	I-protein
(	O
Ab-1	B-protein
)	O
.	O

Stromelysin-1	B-protein
(	O
ST1	B-protein
)	O
was	O
observed	O
in	O
about	O
10	O
%	O
of	O
epithelial	B-cell_type
cells	I-cell_type
in	O
the	O
control	O
groups	O
(	O
cases	O
of	O
fibrocystic	O
and	O
benign	O
proliferative	O
breast	O
disease	O
)	O
,	O
while	O
expression	O
(	O
>	O
10	O
%	O
of	O
expression	O
)	O
was	O
detected	O
in	O
89.7	O
%	O
of	O
tumours	O
.	O

The	O
expression	O
of	O
ST1	B-protein
in	O
carcinoma	O
cells	O
was	O
strongly	O
associated	O
with	O
its	O
presence	O
in	O
the	O
stroma	O
(	O
p	O
<	O
0.001	O
)	O
.	O

A	O
significantly	O
positive	O
correlation	O
was	O
found	O
between	O
ST1	B-protein
expression	O
,	O
and	O
p53	B-protein
tumour	O
suppressor	O
gene	O
product	O
(	O
p	O
=	O
0.004	O
)	O
,	O
and	O
a	O
relationship	O
with	O
c-erbB-2	B-protein
protein	O
and	O
progesterone	O
receptor	O
status	O
was	O
also	O
indicated	O
.	O

These	O
findings	O
suggest	O
that	O
ST1	B-protein
expression	O
in	O
breast	O
cancer	O
tissue	O
is	O
irrespective	O
of	O
the	O
expression	O
of	O
the	O
extracellular	O
matrix	O
component	O
,	O
the	O
proteolytic	B-protein
enzyme	I-protein
cathepsin	B-protein
D	I-protein
and	O
the	O
growth	O
fraction	O
of	O
the	O
tumour	O
,	O
and	O
that	O
it	O
could	O
be	O
a	O
potential	O
new	O
prognostic	O
marker	O
in	O
breast	O
cancer	O
.	O

-DOCSTART-	O

Use	O
of	O
transfected	B-cell_line
liver	I-cell_line
cells	I-cell_line
to	O
evaluate	O
potential	O
mechanisms	O
of	O
alcohol-induced	O
liver	O
injury	O
[	O
see	O
comments	O
]	O

There	O
is	O
increased	O
activity	O
of	O
the	O
proinflammatory	B-protein
cytokine	I-protein
,	O
tumor	B-protein
necrosis	I-protein
factor	I-protein
(	O
TNF	B-protein
)	O
in	O
alcoholic	O
liver	O
disease	O
(	O
ALD	O
)	O
.	O

Hepatic	O
neutrophil	O
infiltration	O
is	O
a	O
principal	O
injurious	O
manifestation	O
of	O
ALD	O
.	O

TNF	B-protein
can	O
induce	O
cellular	O
oxidative	O
injury	O
directly	O
,	O
and	O
indirectly	O
by	O
inducing	O
neutrophil	O
chemotactic	O
factor	O
(	O
IL-8	B-protein
)	O
production	O
by	O
hepatocytes	O
.	O

IL-8	B-protein
activates	O
and	O
chemotactically	O
attracts	O
neutrophils	O
to	O
the	O
liver	O
where	O
they	O
release	O
oxidizing	O
substances	O
.	O

Patients	O
with	O
ALD	O
also	O
have	O
decreased	O
protective	B-protein
factors	I-protein
for	O
cellular	O
oxidative	O
injury	O
.	O

Manganous	B-protein
superoxide	I-protein
dismutase	I-protein
(	O
MnSOD	B-protein
)	O
is	O
an	O
antioxidant	B-protein
protective	I-protein
factor	I-protein
.	O

The	O
objectives	O
of	O
these	O
studies	O
were	O
to	O
investigate	O
mechanisms	O
for	O
induction	O
of	O
an	O
injurious	B-protein
factor	I-protein
(	O
IL-8	B-protein
)	O
and	O
a	O
protective	B-protein
factor	I-protein
(	O
MnSOD	B-protein
)	O
in	O
the	O
HepG2	B-cell_line
human	I-cell_line
hepatoma	I-cell_line
cell	I-cell_line
line	I-cell_line
.	O

In	O
the	O
first	O
set	O
of	O
experiments	O
,	O
IL-8	B-DNA
gene	I-DNA
reporter	I-DNA
constructs	I-DNA
were	O
used	O
to	O
transiently	O
transfect	O
a	O
derivative	O
(	O
MVh2E1-9	B-cell_line
)	O
of	O
the	O
HepG2	B-cell_line
cell	I-cell_line
line	I-cell_line
which	O
expresses	O
P-4502E1	B-protein
and	O
metabolizes	O
ethanol	O
.	O

Inactivation	O
of	O
the	O
NF-kappaB	B-DNA
and	I-DNA
3'NF-IL-6	I-DNA
DNA	I-DNA
binding	I-DNA
sites	I-DNA
decreased	O
IL-8	B-DNA
gene	I-DNA
transcriptional	O
activation	O
in	O
response	O
to	O
TNF	B-protein
while	O
inactivation	O
of	O
the	O
5'NF-IL-6	B-DNA
binding	I-DNA
site	I-DNA
increased	O
IL-8	B-DNA
gene	I-DNA
transcriptional	O
activity	O
in	O
response	O
to	O
TNF	B-protein
.	O

This	O
system	O
may	O
be	O
useful	O
to	O
assess	O
the	O
effects	O
of	O
ethanol	O
on	O
TNF	B-protein
-induced	O
hepatocyte	O
IL-8	B-protein
production	O
.	O

In	O
the	O
second	O
set	O
of	O
experiments	O
,	O
HepG2	B-cell_line
cells	I-cell_line
were	O
cultured	O
in	O
25	O
to	O
100	O
mmol	O
concentrations	O
of	O
ethanol	O
.	O

Both	O
TNF	B-protein
and	O
ethanol	O
increased	O
HepG2	B-cell_line
cell	I-cell_line
MnSOD	B-protein
activity	O
in	O
short-term	O
(	O
72	O
hr	O
)	O
cultures	O
with	O
ethanol	O
.	O

However	O
,	O
after	O
long-term	O
(	O
10	O
weeks	O
)	O
culture	O
with	O
ethanol	O
,	O
there	O
was	O
no	O
induction	O
of	O
MnSOD	B-protein
by	O
ethanol	O
and	O
there	O
was	O
a	O
diminished	O
induction	O
of	O
MnSOD	B-protein
in	O
response	O
to	O
TNF	B-protein
.	O

Further	O
studies	O
are	O
needed	O
to	O
assess	O
the	O
effect	O
of	O
this	O
diminished	O
induction	O
of	O
MnSOD	B-protein
with	O
chronic	O
ethanol	O
culture	O
on	O
HepG2	B-cell_line
cell	I-cell_line
susceptibility	O
to	O
TNF	B-cell_line
cytotoxicity	I-cell_line
.	O

We	O
conclude	O
that	O
transfected	O
liver	B-cell_line
cell	I-cell_line
lines	I-cell_line
can	O
be	O
used	O
to	O
evaluate	O
mechanisms	O
for	O
increased	O
injurious	B-protein
factors	I-protein
and	O
decreased	O
protective	B-protein
factors	I-protein
in	O
alcoholic	O
liver	O
injury	O
.	O

-DOCSTART-	O

Tissue	O
factor	O
transcription	O
driven	O
by	O
Egr-1	B-protein
is	O
a	O
critical	O
mechanism	O
of	O
murine	B-protein
pulmonary	I-protein
fibrin	I-protein
deposition	O
in	O
hypoxia	O
.	O

Local	O
hypoxemia	O
and	O
stasis	O
trigger	O
thrombosis	O
.	O

We	O
have	O
demonstrated	O
previously	O
that	O
in	O
a	O
murine	O
model	O
of	O
normobaric	O
hypoxia	O
pulmonary	O
fibrin	B-protein
deposition	O
is	O
a	O
result	O
of	O
expression	O
of	O
tissue	B-protein
factor	I-protein
,	O
especially	O
in	O
oxygen-deprived	B-protein
mononuclear	I-protein
phagocytes	I-protein
(	O
MPs	B-protein
)	O
.	O

We	O
now	O
show	O
that	O
transcription	B-protein
factor	I-protein
early-growth-response	I-protein
gene	I-protein
product	I-protein
(	O
Egr-1	B-protein
)	O
is	O
rapidly	O
activated	O
in	O
hypoxia	O
,	O
both	O
in	O
vitro	O
and	O
in	O
vivo	O
,	O
and	O
is	O
responsible	O
for	O
transcription	O
and	O
expression	O
of	O
tissue	B-protein
factor	I-protein
in	O
hypoxic	O
lung	O
.	O

MPs	B-protein
and	O
HeLa	B-cell_line
cells	I-cell_line
subjected	O
to	O
hypoxia	O
(	O
pO2	O
approximately	O
13	O
torr	O
)	O
had	O
increased	O
levels	O
of	O
tissue	B-RNA
factor	I-RNA
transcripts	I-RNA
(	O
approximately	O
18-fold	O
)	O
and	O
an	O
increased	O
rate	O
of	O
transcription	O
(	O
approximately	O
15-fold	O
)	O
,	O
based	O
on	O
nuclear	O
run-on	O
analysis	O
.	O

Gel-shift	O
analysis	O
of	O
nuclear	O
extracts	O
from	O
hypoxic	B-protein
MPs	I-protein
and	O
HeLa	B-cell_line
cells	I-cell_line
demonstrated	O
increased	O
DNA-binding	O
activity	O
at	O
the	O
serum	B-DNA
response	I-DNA
region	I-DNA
(	O
SRR	O
;	O
-111/+14	O
bp	O
)	O
of	O
the	O
tissue	B-DNA
factor	I-DNA
promoter	I-DNA
at	O
Egr-1	B-DNA
motifs	I-DNA
.	O

Using	O
32P-labeled	O
Egr	O
consensus	O
oligonucleotide	O
,	O
we	O
observed	O
induction	O
of	O
DNA-binding	O
activity	O
in	O
nuclear	O
extracts	O
from	O
hypoxic	O
lung	O
and	O
HeLa	B-cell_line
cells	I-cell_line
because	O
of	O
activation	O
of	O
Egr-1	B-protein
,	O
by	O
means	O
of	O
supershift	O
analysis	O
.	O

Transient	O
transfection	O
of	O
HeLa	B-cell_line
cells	I-cell_line
with	O
chimeric	B-DNA
plasmids	I-DNA
containing	O
wild-type	B-protein
or	I-protein
mutant	I-protein
SRR	I-protein
from	O
the	O
tissue	B-DNA
factor	I-DNA
promoter	I-DNA
showed	O
that	O
intact	B-DNA
Sp1	I-DNA
sites	I-DNA
are	O
necessary	O
for	O
basal	O
promoter	O
activity	O
,	O
whereas	O
the	O
integrity	O
of	O
Egr-1	B-DNA
sites	I-DNA
was	O
required	O
for	O
hypoxia-enhanced	O
expression	O
.	O

A	O
central	O
role	O
for	O
Egr-1	B-protein
in	O
hypoxia-mediated	O
tissue	O
factor	O
expression	O
was	O
confirmed	O
by	O
experiments	O
with	O
homozygous	O
Egr-1	B-protein
null	O
mice	O
;	O
wild-type	O
mice	O
subjected	O
to	O
oxygen	B-protein
deprivation	I-protein
expressed	I-protein
tissue	I-protein
factor	I-protein
and	O
showed	O
fibrin	B-protein
deposition	O
,	O
but	O
hypoxic	O
homozygous	O
Egr-1	B-protein
null	O
mice	O
displayed	O
neither	O
tissue	B-protein
factor	I-protein
nor	O
fibrin	B-protein
.	O

These	O
data	O
delineate	O
a	O
novel	O
biology	O
for	O
hypoxia-induced	O
fibrin	B-protein
deposition	O
,	O
in	O
which	O
oxygen	O
deprivation-induced	O
activation	O
of	O
Egr-1	B-protein
,	O
resulting	O
in	O
expression	O
of	O
tissue	B-protein
factor	I-protein
,	O
has	O
an	O
unexpected	O
and	O
central	O
role	O
.	O

-DOCSTART-	O

Kinetics	O
of	O
cytokine	O
and	O
NFAT	O
gene	O
expression	O
in	O
human	B-cell_line
interleukin-2-dependent	I-cell_line
T	I-cell_line
lymphoblasts	I-cell_line
stimulated	O
via	O
T-cell	B-protein
receptor	I-protein
.	O

T	B-cell_type
cells	I-cell_type
respond	O
to	O
mitogenic	O
or	O
antigenic	O
stimulation	O
by	O
proliferation	O
and	O
by	O
turning	O
on	O
cytokine	O
gene	O
expression	O
.	O

Here	O
we	O
have	O
analysed	O
the	O
kinetics	O
and	O
nature	O
of	O
cytokine	O
production	O
in	O
human	B-cell_type
peripheral	I-cell_type
blood-derived	I-cell_type
T	I-cell_type
lymphoblasts	I-cell_type
stimulated	O
with	O
anti-CD3	B-protein
antibodies	I-protein
or	O
Lens	B-protein
culinaris	I-protein
lectin	I-protein
(	O
LCL	B-protein
)	O
.	O

T	B-cell_type
cells	I-cell_type
were	O
purified	O
from	O
peripheral	B-cell_type
blood	I-cell_type
mononuclear	I-cell_type
cells	I-cell_type
(	O
PBMC	B-cell_type
)	O
and	O
primarily	O
activated	O
with	O
anti-CD3	B-protein
antibodies	I-protein
and	O
cultured	O
in	O
the	O
presence	O
of	O
interleukin-2	B-protein
(	O
IL-2	B-protein
)	O
.	O

Anti-CD3-restimulated	O
T	B-cell_type
cells	I-cell_type
(	O
mainly	O
CD8+	O
)	O
produced	O
IL-2	B-protein
,	O
interferon-gamma	B-protein
(	O
IFN-gamma	B-protein
)	O
and	O
tumour	B-protein
necrosis	I-protein
factor-alpha	I-protein
(	O
TNF-alpha	B-protein
)	O
and	O
low	O
levels	O
of	O
IL-4	B-RNA
and	I-RNA
IL-10	I-RNA
transcripts	I-RNA
and	O
proteins	O
.	O

No	O
IL-6	B-DNA
gene	I-DNA
expression	O
was	O
observed	O
.	O

In	O
LCL-stimulated	B-cell_type
cells	I-cell_type
the	O
cytokine	O
production	O
pattern	O
was	O
very	O
similar	O
.	O

Steady-state	O
mRNA	O
levels	O
of	O
IL-2	B-protein
,	O
IL-10	B-protein
and	O
IFN-gamma	B-protein
peaked	O
at	O
3	O
hr	O
after	O
anti-CD3	O
stimulation	O
and	O
declined	O
rapidly	O
thereafter	O
.	O

The	O
kinetics	O
of	O
TNF-alpha	B-RNA
mRNA	I-RNA
expression	O
was	O
faster	O
,	O
being	O
at	O
its	O
peak	O
level	O
1	O
hr	O
after	O
stimulation	O
.	O

Anti-CD3-stimulated	O
IL-2	B-DNA
gene	I-DNA
expression	O
was	O
down-regulated	O
by	O
protein	O
synthesis	O
inhibitor	O
,	O
whereas	O
IL-10	B-DNA
,	I-DNA
IFN-gamma	I-DNA
and	I-DNA
TNF-alpha	I-DNA
genes	I-DNA
were	O
readily	O
induced	O
independent	O
of	O
ongoing	O
protein	O
synthesis	O
.	O

T-cell	B-protein
receptor	I-protein
stimulation	O
also	O
induced	O
a	O
very	O
rapid	O
expression	O
of	O
c-jun	B-DNA
,	I-DNA
c-fos	I-DNA
and	I-DNA
NFATc1	I-DNA
(	I-DNA
NFATc	I-DNA
)	I-DNA
genes	I-DNA
,	O
the	O
gene	O
products	O
of	O
which	O
are	O
involved	O
in	O
cytokine	O
gene	O
expression	O
.	O

In	O
conclusion	O
,	O
the	O
cytokines	B-protein
synthesized	O
by	O
IL-2-dependent	B-cell_type
T	I-cell_type
cells	I-cell_type
were	O
predominantly	O
IL-2	B-protein
,	O
IFN-gamma	B-protein
and	O
TNF-alpha	B-protein
.	O

-DOCSTART-	O

An	O
animal	O
model	O
to	O
study	O
local	O
oxidation	O
of	O
LDL	O
and	O
its	O
biological	O
effects	O
in	O
the	O
arterial	O
wall	O
.	O

Oxidized	O
LDL	O
(	O
oxLDL	O
)	O
is	O
present	O
in	O
atherosclerotic	O
lesions	O
and	O
is	O
believed	O
to	O
play	O
a	O
key	O
role	O
in	O
atherogenesis	O
.	O

Mainly	O
on	O
the	O
basis	O
of	O
cell	O
culture	O
studies	O
,	O
oxLDL	O
has	O
been	O
shown	O
to	O
produce	O
many	O
biological	O
effects	O
that	O
influence	O
the	O
atherosclerotic	O
process	O
.	O

To	O
study	O
LDL	O
oxidation	O
in	O
vivo	O
,	O
we	O
have	O
established	O
a	O
model	O
in	O
which	O
Sprague-Dawley	O
rats	O
are	O
given	O
a	O
single	O
injection	O
of	O
unmodified	O
human	O
LDL	O
(	O
>	O
or	O
=	O
4	O
mg/kg	O
body	O
weight	O
)	O
.	O

Within	O
6	O
hours	O
,	O
an	O
accumulation	O
of	O
apolipoprotein	O
B	O
and	O
epitopes	B-protein
present	O
on	O
oxLDL	O
are	O
detected	O
in	O
the	O
arterial	O
endothelium	O
and	O
media	O
.	O

The	O
presence	O
of	O
oxLDL	O
is	O
associated	O
with	O
activation	O
of	O
the	O
transcription	B-protein
factor	I-protein
nuclear	B-protein
factor-kappaB	I-protein
in	O
the	O
endothelium	O
as	O
well	O
as	O
endothelial	O
expression	O
of	O
intercellular	B-protein
adhesion	I-protein
molecule-1	I-protein
.	O

Injection	O
of	O
LDL	O
enriched	O
with	O
the	O
antioxidant	O
probucol	O
resulted	O
in	O
arterial	O
accumulation	O
of	O
apolipoprotein	O
B	O
,	O
but	O
the	O
expression	O
of	O
oxLDL-specific	O
epitopes	O
was	O
reduced	O
at	O
24	O
hours	O
.	O

Thus	O
,	O
this	O
simple	O
model	O
has	O
the	O
potential	O
to	O
analyze	O
the	O
mechanisms	O
behind	O
and	O
biological	O
effects	O
of	O
LDL	O
oxidation	O
in	O
vivo	O
.	O

-DOCSTART-	O

Molecular	O
mechanisms	O
of	O
promoter	O
regulation	O
of	O
the	O
gp34	B-DNA
gene	I-DNA
that	O
is	O
trans-activated	O
by	O
an	O
oncoprotein	B-protein
Tax	I-protein
of	O
human	O
T	O
cell	O
leukemia	O
virus	O
type	O
I	O
.	O

We	O
investigated	O
the	O
molecular	O
mechanism	O
of	O
transcriptional	O
activation	O
of	O
the	O
gp34	B-DNA
gene	I-DNA
by	O
the	O
Tax	B-protein
oncoprotein	I-protein
of	O
human	O
T	O
cell	O
leukemia	O
virus	O
type	O
I	O
(	O
HTLV-I	O
)	O
.	O

gp34	B-protein
is	O
a	O
type	B-protein
II	I-protein
transmembrane	I-protein
molecule	I-protein
belonging	O
to	O
the	O
tumor	O
necrosis	O
factor	O
family	O
and	O
is	O
constitutively	O
expressed	O
on	O
HTLV-I-producing	B-cell_type
cells	I-cell_type
but	O
not	O
normal	B-cell_type
resting	I-cell_type
T	I-cell_type
cells	I-cell_type
.	O

The	O
transcriptional	O
regulatory	O
region	O
of	O
the	O
gp34	B-DNA
gene	I-DNA
was	O
activated	O
by	O
HTLV-I	O
Tax	B-protein
in	O
the	O
human	B-cell_line
T	I-cell_line
cell	I-cell_line
line	I-cell_line
Jurkat	I-cell_line
,	O
in	O
which	O
endogenous	O
gp34	B-protein
is	O
induced	O
by	O
Tax	B-protein
.	O

Sequence	O
analysis	O
demonstrated	O
that	O
two	O
NF-kappaB-like	B-DNA
elements	I-DNA
(	O
1	O
and	O
2	O
)	O
were	O
present	O
in	O
the	O
regulatory	B-DNA
region	I-DNA
.	O

Both	O
NF-kappaB-like	B-DNA
elements	I-DNA
were	O
able	O
to	O
bind	O
to	O
NF-kappaB	B-protein
or	O
its	O
related	O
factor	O
(	O
s	O
)	O
in	O
a	O
Tax	B-protein
-dependent	O
manner	O
.	O

Chloramphenicol	B-protein
acetyltransferase	I-protein
assays	O
indicated	O
that	O
NF-kappaB-like	B-DNA
element	I-DNA
1	I-DNA
was	O
Tax	B-protein
-responsive	O
,	O
although	O
the	O
activity	O
was	O
lower	O
than	O
that	O
the	O
native	B-DNA
promoter	I-DNA
.	O

NF-kappaB	B-protein
-like	O
element	O
2	O
elevated	O
promoter	O
activity	O
when	O
combined	O
with	O
NF-kappaB-like	B-DNA
element	I-DNA
1	I-DNA
,	O
indicating	O
cooperative	O
function	O
of	O
the	O
elements	O
for	O
maximum	O
promoter	O
function	O
.	O

Unlike	O
typical	O
NF-kappaB	B-DNA
elements	I-DNA
,	O
the	O
NF-kappaB-like	B-DNA
elements	I-DNA
in	O
gp34	B-protein
were	O
not	O
activated	O
by	O
treatment	O
of	O
Jurkat	B-cell_line
cells	I-cell_line
with	O
phorbol	O
ester	O
despite	O
induction	O
of	O
the	O
NF-kappaB	B-protein
-like	O
binding	O
activity	O
.	O

Chloramphenicol	B-protein
acetyltransferase	I-protein
reporter	O
assays	O
using	O
the	O
region	O
upstream	O
of	O
the	O
NF-kappaB-like	B-DNA
elements	I-DNA
identified	O
an	O
upstream	O
region	O
that	O
reduced	O
transcription	O
from	O
cognate	B-DNA
and	I-DNA
noncognate	I-DNA
core	I-DNA
promoters	I-DNA
in	O
a	O
Tax-independent	O
manner	O
.	O

Our	O
results	O
imply	O
complex	O
regulation	O
of	O
expression	O
of	O
the	O
gp34	B-DNA
gene	I-DNA
and	O
suggest	O
implication	O
of	O
gp34	B-protein
in	O
proliferation	O
of	O
HTLV-I	O
infected	O
T	O
cells	O
.	O

-DOCSTART-	O

beta-Amyloid	B-protein
fibrils	I-protein
activate	O
parallel	O
mitogen-activated	B-protein
protein	I-protein
kinase	I-protein
pathways	O
in	O
microglia	B-cell_type
and	O
THP1	B-cell_line
monocytes	I-cell_line
.	O

The	O
senile	O
plaques	O
of	O
Alzheimer	O
's	O
disease	O
are	O
foci	O
of	O
local	O
inflammatory	O
responses	O
,	O
as	O
evidenced	O
by	O
the	O
presence	O
of	O
acute	B-protein
phase	I-protein
proteins	I-protein
and	O
oxidative	O
damage	O
.	O

Fibrillar	B-protein
forms	I-protein
of	O
beta-amyloid	B-protein
(	O
Abeta	B-protein
)	O
,	O
which	O
are	O
the	O
primary	O
constituents	O
of	O
senile	O
plaques	O
,	O
have	O
been	O
shown	O
to	O
activate	O
tyrosine	O
kinase-dependent	O
signal	O
transduction	O
cascades	O
,	O
resulting	O
in	O
inflammatory	O
responses	O
in	O
microglia	B-cell_type
.	O

However	O
,	O
the	O
downstream	O
signaling	O
pathways	O
mediating	O
Abeta	B-protein
-induced	O
inflammatory	O
events	O
are	O
not	O
well	O
characterized	O
.	O

We	O
report	O
that	O
exposure	O
of	O
primary	B-cell_type
rat	I-cell_type
microglia	I-cell_type
and	O
human	B-cell_line
THP1	I-cell_line
monocytes	I-cell_line
to	O
fibrillar	B-protein
Abeta	I-protein
results	O
in	O
the	O
tyrosine	B-protein
kinase	I-protein
-dependent	O
activation	O
of	O
two	O
parallel	O
signal	O
transduction	O
cascades	O
involving	O
members	O
of	O
the	O
mitogen-activated	B-protein
protein	I-protein
kinase	I-protein
(	I-protein
MAPK	I-protein
)	I-protein
superfamily	I-protein
.	O

Abeta	B-protein
stimulated	O
the	O
rapid	O
,	O
transient	O
activation	O
of	O
extracellular	B-protein
signal-regulated	I-protein
kinase	I-protein
1	I-protein
(	O
ERK1	B-protein
)	O
and	O
ERK2	B-protein
in	O
microglia	B-cell_type
and	O
ERK2	B-protein
in	O
THP1	B-cell_line
monocytes	I-cell_line
.	O

A	O
second	O
superfamily	B-protein
member	I-protein
,	O
p38	B-protein
MAPK	I-protein
,	O
was	O
also	O
activated	O
with	O
similar	O
kinetics	O
.	O

Scavenger	B-protein
receptor	I-protein
and	O
receptor	B-protein
for	I-protein
advanced	I-protein
glycated	I-protein
end	I-protein
products	I-protein
(	O
RAGE	B-protein
)	O
ligands	O
failed	O
to	O
activate	O
ERK	B-protein
and	O
p38	B-protein
MAPK	I-protein
in	O
the	O
absence	O
of	O
significant	O
increases	O
in	O
protein	O
tyrosine	O
phosphorylation	O
,	O
demonstrating	O
that	O
scavenger	B-protein
receptors	I-protein
and	O
RAGE	B-protein
are	O
not	O
linked	O
to	O
these	O
pathways	O
.	O

Importantly	O
,	O
the	O
stress-activated	B-protein
protein	I-protein
kinases	I-protein
(	O
SAPKs	B-protein
)	O
were	O
not	O
significantly	O
activated	O
in	O
response	O
to	O
Abeta	B-protein
.	O

Downstream	O
effectors	O
of	O
the	O
MAPK	O
signal	O
transduction	O
cascades	O
include	O
MAPKAP	B-protein
kinases	I-protein
,	O
such	O
as	O
RSK1	B-protein
and	O
RSK2	B-protein
,	O
as	O
well	O
as	O
transcription	B-protein
factors	I-protein
.	O

Exposure	O
of	O
microglia	B-cell_type
and	O
THP1	B-cell_line
monocytes	I-cell_line
to	O
Abeta	B-protein
resulted	O
in	O
the	O
activation	O
of	O
RSK1	B-protein
and	O
RSK2	B-protein
and	O
phosphorylation	O
of	O
cAMP	B-protein
response	I-protein
element-binding	I-protein
protein	I-protein
at	O
Ser133	O
,	O
providing	O
a	O
mechanism	O
for	O
Abeta	B-protein
-induced	O
changes	O
in	O
gene	O
expression	O

-DOCSTART-	O

Regulation	O
of	O
cellular	B-protein
retinoic	I-protein
acid	I-protein
binding	I-protein
protein	I-protein
(	O
CRABP	B-protein
II	I-protein
)	O
during	O
human	B-cell_type
monocyte	I-cell_type
differentiation	O
in	O
vitro	O
.	O

Cellular	B-protein
retinoic	I-protein
acid	I-protein
binding	I-protein
proteins	I-protein
(	O
CRABP	B-protein
)	O
are	O
low	B-protein
molecular	I-protein
weight	I-protein
proteins	I-protein
whose	O
precise	O
function	O
remains	O
unknown	O
.	O

They	O
bind	O
retinoids	O
and	O
may	O
thereby	O
modulate	O
the	O
intracellular	O
steady-state	O
concentration	O
of	O
retinoids	O
.	O

Whereas	O
CRABP	B-protein
I	I-protein
is	O
ubiquitously	O
expressed	O
,	O
CRABP	B-protein
II	I-protein
is	O
mainly	O
detected	O
in	O
various	O
cell	O
types	O
of	O
the	O
skin	O
.	O

By	O
representative	O
difference	O
analysis	O
we	O
found	O
that	O
CRABP	B-protein
II	I-protein
is	O
also	O
strongly	O
expressed	O
in	O
human	B-cell_type
monocyte-derived	I-cell_type
macrophages	I-cell_type
(	O
MAC	B-cell_type
)	O
but	O
not	O
in	O
freshly	B-cell_type
isolated	I-cell_type
monocytes	I-cell_type
(	O
MO	B-cell_type
)	O
.	O

The	O
CRABP	B-RNA
II	I-RNA
mRNA	I-RNA
was	O
gradually	O
upregulated	O
during	O
differentiation	O
from	O
MO	B-cell_type
to	O
MAC	B-cell_type
in	O
the	O
presence	O
of	O
2	O
%	O
serum	O
.	O

Adherence	O
,	O
which	O
is	O
important	O
for	O
MO	B-cell_type
differentiation	O
,	O
induced	O
CRABP	B-protein
II	I-protein
expression	O
,	O
but	O
the	O
addition	O
of	O
10	O
(	O
-7	O
)	O
M	O
retinoic	O
acid	O
inhibited	O
the	O
upregulation	O
of	O
CRABP	B-protein
II	I-protein
expression	O
during	O
MO/MAC	O
differentiation	O
.	O

As	O
MO	B-cell_type
can	O
differentiate	O
along	O
the	O
classical	O
pathway	O
not	O
only	O
to	O
MAC	B-cell_type
but	O
also	O
to	O
dendritic	O
cells	O
we	O
analyzed	O
the	O
expression	O
of	O
CRABP	B-protein
II	I-protein
in	O
MO-derived	B-cell_type
dendritic	I-cell_type
cells	I-cell_type
cultured	O
with	O
10	O
%	O
FCS	B-protein
,	O
IL-4	B-protein
,	O
and	O
GM-CSF	B-protein
.	O

In	O
contrast	O
to	O
MAC	B-cell_type
,	O
MO-derived	B-cell_type
dendritic	I-cell_type
cells	I-cell_type
showed	O
an	O
extremely	O
low	O
expression	O
of	O
CRABP	B-protein
II	I-protein
.	O

From	O
these	O
results	O
we	O
conclude	O
(	O
1	O
)	O
that	O
the	O
availability	O
and	O
the	O
metabolism	O
of	O
retinoids	O
may	O
be	O
different	O
in	O
MAC	B-cell_type
compared	O
to	O
MO	B-cell_type
and	O
dendritic	B-cell_type
cells	I-cell_type
and	O
(	O
2	O
)	O
that	O
this	O
may	O
influence	O
differentiation	O
and	O
activation	O
of	O
those	O
cells	O
.	O

-DOCSTART-	O

Transcription	B-protein
factor	I-protein
B-cell-specific	I-protein
activator	I-protein
protein	I-protein
(	O
BSAP	B-protein
)	O
is	O
differentially	O
expressed	O
in	O
B	B-cell_type
cells	I-cell_type
and	O
in	O
subsets	O
of	O
B-cell	O
lymphomas	O
.	O

The	O
paired	B-DNA
box	I-DNA
containing	I-DNA
gene	I-DNA
PAX-5	B-DNA
encodes	O
the	O
transcription	B-protein
factor	I-protein
BSAP	B-protein
(	O
B-cell-specific	B-protein
activator	I-protein
protein	I-protein
)	O
,	O
which	O
plays	O
a	O
key	O
role	O
in	O
B-lymphocyte	O
development	O
.	O

Despite	O
its	O
known	O
involvement	O
in	O
a	O
rare	O
subtype	O
of	O
non-Hodgkin	O
's	O
lymphoma	O
(	O
NHL	O
)	O
,	O
a	O
detailed	O
examination	O
of	O
BSAP	B-protein
expression	O
in	O
NHL	O
has	O
not	O
been	O
previously	O
reported	O
.	O

In	O
this	O
study	O
,	O
we	O
analyzed	O
normal	O
and	O
malignant	O
lymphoid	O
tissues	O
and	O
cell	B-cell_line
lines	I-cell_line
,	O
including	O
102	O
cases	O
of	O
B-cell	O
NHL	O
,	O
23	O
cases	O
of	O
T-	O
and	O
null-cell	O
NHL	O
,	O
and	O
18	O
cases	O
of	O
Hodgkin	O
's	O
disease	O
.	O

Normal	O
lymphoid	O
tissues	O
showed	O
strong	O
nuclear	O
BSAP	B-protein
expression	O
in	O
mantle	B-cell_type
zone	I-cell_type
B	I-cell_type
cells	I-cell_type
,	O
less	O
intense	O
reactivity	O
in	O
follicular	B-cell_type
center	I-cell_type
B	I-cell_type
cells	I-cell_type
,	O
and	O
no	O
expression	O
in	O
cells	O
of	O
the	O
T-cell-rich	O
zones	O
.	O

Monocytoid	B-cell_type
B	I-cell_type
cells	I-cell_type
showed	O
weak	O
expression	O
,	O
whereas	O
plasma	B-cell_type
cells	I-cell_type
and	O
extrafollicular	B-cell_type
large	I-cell_type
transformed	I-cell_type
B	I-cell_type
cells	I-cell_type
were	O
negative	O
.	O

Of	O
the	O
102	O
B-cell	O
NHLs	O
,	O
83	O
(	O
81	O
%	O
)	O
demonstrated	O
BSAP	B-protein
expression	O
.	O

All	O
of	O
the	O
13	O
(	O
100	O
%	O
)	O
B-cell	O
chronic	O
lymphocytic	O
leukemias	O
(	O
B-CLLs	O
)	O
,	O
21	O
of	O
(	O
100	O
%	O
)	O
mantle	B-cell_type
cells	I-cell_type
(	O
MCLs	B-cell_type
)	O
,	O
and	O
20	O
of	O
21	O
(	O
95	O
%	O
)	O
follicular	O
lymphomas	O
(	O
FLs	O
)	O
were	O
positive	O
.	O

Moderate	O
staining	O
intensities	O
were	O
found	O
in	O
most	O
B-CLL	O
and	O
FL	O
cases	O
,	O
whereas	O
most	O
MCLs	B-cell_type
showed	O
strong	O
reactions	O
,	O
paralleling	O
the	O
strong	O
reactivity	O
of	O
nonmalignant	B-cell_type
mantle	I-cell_type
cells	I-cell_type
.	O

Eight	O
of	O
12	O
(	O
67	O
%	O
)	O
marginal	O
zone	O
lymphoma	O
cases	O
showed	O
negative	O
or	O
low	O
BSAP	B-protein
levels	O
,	O
and	O
17	O
of	O
24	O
(	O
71	O
%	O
)	O
large	O
B-cell	O
lymphomas	O
displayed	O
moderate	O
to	O
strong	O
expression	O
.	O

None	O
of	O
the	O
23	O
T-	O
and	O
null-cell	O
lymphomas	O
reacted	O
with	O
the	O
BSAP	B-protein
antisera	O
,	O
whereas	O
in	O
Hodgkin	O
's	O
disease	O
,	O
2	O
of	O
4	O
(	O
50	O
%	O
)	O
nodular	O
lymphocytic	O
predominance	O
and	O
5	O
of	O
14	O
(	O
36	O
%	O
)	O
classical	O
cases	O
showed	O
weak	O
nuclear	O
or	O
nucleolar	O
BSAP	B-protein
reactions	O
in	O
a	O
fraction	O
of	O
the	O
tumor	B-cell_type
cells	I-cell_type
.	O

Western	O
blot	O
analysis	O
showed	O
a	O
52-kD	O
BSAP	B-protein
band	O
in	O
B-cell	B-cell_line
lines	I-cell_line
,	O
but	O
not	O
in	O
non-B-cell	B-cell_line
or	I-cell_line
plasma	I-cell_line
cell	I-cell_line
lines	I-cell_line
.	O

We	O
conclude	O
that	O
BSAP	B-protein
expression	O
is	O
largely	O
restricted	O
to	O
lymphomas	O
of	O
B-cell	B-cell_type
lineage	I-cell_type
and	O
that	O
BSAP	B-protein
expression	O
varies	O
in	O
B-cell	B-cell_type
subsets	I-cell_type
and	O
subtypes	O
of	O
B-cell	O
NHL	O
.	O

The	O
high	O
levels	O
of	O
BSAP	B-protein
,	O
especially	O
those	O
found	O
in	O
large-cell	B-cell_type
lymphomas	I-cell_type
and	O
in	O
some	O
follicular	B-cell_type
lymphomas	I-cell_type
,	O
may	O
be	O
a	O
consequence	O
of	O
deregulated	O
gene	O
expression	O
and	O
suggest	O
a	O
possible	O
involvement	O
of	O
PAX-5	B-DNA
in	O
certain	O
B-cell	O
malignancies	O
.	O

This	O
is	O
a	O
US	O
government	O
work	O
.	O

There	O
are	O
no	O
restrictions	O
on	O
its	O
use	O
.	O

-DOCSTART-	O

Mitogen	O
and	O
growth	O
factor-induced	O
activation	O
of	O
a	O
STAT-like	B-protein
molecule	I-protein
in	O
channel	B-cell_type
catfish	I-cell_type
lymphoid	I-cell_type
cells	I-cell_type
.	O

This	O
article	O
describes	O
the	O
identification	O
of	O
a	O
putative	B-protein
STAT	I-protein
molecule	I-protein
in	O
the	O
channel	O
catfish	O
(	O
Ictalurus	O
punctatus	O
)	O
,	O
the	O
first	O
report	O
of	O
such	O
a	O
molecule	O
in	O
a	O
'lower	O
'	O
vertebrate	O
.	O

A	O
monoclonal	B-protein
antibody	I-protein
against	O
human	B-protein
STAT6	I-protein
recognizes	O
an	O
approximately	O
100	B-protein
kDa	I-protein
molecule	I-protein
that	O
becomes	O
activated	O
and	O
translocates	O
to	O
the	O
nucleus	O
upon	O
both	O
growth	O
factor	O
and	O
mitogen	O
stimulation	O
of	O
catfish	O
leukocytes	O
.	O

This	O
presumed	O
catfish	B-protein
STAT	I-protein
binds	O
the	O
mammalian	B-DNA
interferon-gamma	I-DNA
activation	I-DNA
site	I-DNA
,	O
a	O
known	O
motif	O
of	O
mammalian	O
STAT	O
binding	O
,	O
as	O
shown	O
by	O
electromobility	O
shift	O
assays	O
.	O

Purification	O
of	O
the	O
proteins	O
present	O
in	O
these	O
DNA	B-protein
complexes	I-protein
confirms	O
that	O
the	O
catfish	O
reactive	O
molecule	O
binds	O
to	O
the	O
interferon-gamma	B-DNA
activation	I-DNA
site	I-DNA
sequence	I-DNA
.	O

These	O
results	O
suggest	O
that	O
STAT	B-protein
molecules	I-protein
have	O
been	O
highly	O
conserved	O
in	O
vertebrate	O
evolution	O
.	O

-DOCSTART-	O

Isolation	O
and	O
analysis	O
of	O
a	O
T	B-cell_line
cell	I-cell_line
clone	I-cell_line
variant	I-cell_line
exhibiting	O
constitutively	O
phosphorylated	B-protein
Ser133	I-protein
cAMP	I-protein
response	I-protein
element-binding	I-protein
protein	I-protein
.	O

In	O
driving	O
T	O
cell	O
proliferation	O
,	O
IL-2	B-protein
stimulates	O
a	O
new	O
program	O
of	O
gene	O
expression	O
that	O
includes	O
proliferating	B-protein
cell	I-protein
nuclear	I-protein
antigen	I-protein
(	O
PCNA	B-protein
)	O
,	O
a	O
requisite	O
processivity	O
factor	O
for	O
DNA	B-protein
polymerase	I-protein
delta	I-protein
.	O

PCNA	B-protein
transcription	O
is	O
regulated	O
in	O
part	O
through	O
tandem	O
CRE	B-DNA
sequences	I-DNA
in	O
the	O
promoter	B-protein
and	I-protein
CRE	I-protein
binding	I-protein
proteins	I-protein
;	O
IL-2	B-protein
stimulates	O
CREB	O
phosphorylation	O
in	O
the	O
resting	B-cell_line
cloned	I-cell_line
T	I-cell_line
lymphocyte	I-cell_line
,	O
L2	B-cell_line
.	O

After	O
culturing	O
L2	B-cell_line
cells	I-cell_line
for	O
greater	O
than	O
91	O
days	O
,	O
we	O
consistently	O
isolate	O
a	O
stable	O
variant	O
that	O
exhibits	O
constitutive	O
CREB	O
phosphorylation	O
.	O

L2	B-cell_line
and	I-cell_line
L2	I-cell_line
variant	I-cell_line
cells	I-cell_line
were	O
tested	O
for	O
IL-2	B-protein
responsiveness	O
and	O
rapamycin	O
sensitivity	O
with	O
respect	O
to	O
specific	O
kinase	O
activity	O
,	O
PCNA	O
expression	O
and	O
proliferation	O
.	O

In	O
L2	B-cell_line
cells	I-cell_line
,	O
IL-2	B-protein
stimulated	O
and	O
rapamycin	O
inhibited	O
the	O
following	O
:	O
cAMP-independent	O
CREB	B-protein
kinase	I-protein
activity	O
,	O
PCNA	O
expression	O
and	O
proliferation	O
.	O

In	O
L2	B-cell_line
variant	I-cell_line
cells	I-cell_line
,	O
CREB	B-protein
kinase	I-protein
activity	O
was	O
constitutively	O
high	O
;	O
IL-2	B-protein
stimulated	O
and	O
rapamycin	O
blocked	O
PCNA	O
expression	O
and	O
proliferation	O
.	O

These	O
results	O
indicate	O
that	O
IL-2	B-protein
induces	O
a	O
rapamycin-sensitive	O
,	O
cAMP-independent	O
CREB	B-protein
kinase	I-protein
activity	O
in	O
L2	B-cell_line
cells	I-cell_line
.	O

However	O
,	O
phosphorylation	O
of	O
CREB	B-protein
alone	O
is	O
not	O
sufficient	O
to	O
drive	O
PCNA	B-protein
expression	O
and	O
L2	O
cell	O
proliferation	O
in	O
the	O
absence	O
of	O
IL-2	B-protein
.	O

-DOCSTART-	O

Serotonin	O
derivative	O
,	O
N-	O
(	O
p-coumaroyl	O
)	O
serotonin	O
,	O
inhibits	O
the	O
production	O
of	O
TNF-alpha	B-protein
,	O
IL-1alpha	B-protein
,	O
IL-1beta	B-protein
,	O
and	O
IL-6	B-protein
by	O
endotoxin-stimulated	B-cell_type
human	I-cell_type
blood	I-cell_type
monocytes	I-cell_type
.	O

We	O
have	O
reported	O
that	O
N-	O
(	O
p-coumaroyl	O
)	O
serotonin	O
(	O
CS	O
)	O
and	O
its	O
derivatives	O
with	O
antioxidative	O
activity	O
are	O
present	O
in	O
safflower	O
seeds	O
.	O

As	O
reactive	O
oxygen	O
species	O
(	O
ROS	O
)	O
are	O
implicated	O
in	O
the	O
signaling	O
of	O
lipopolysaccharide	O
(	O
LPS	O
)	O
,	O
we	O
examined	O
whether	O
CS	O
has	O
a	O
suppressive	O
effect	O
on	O
inflammatory	B-protein
cytokine	I-protein
generation	O
from	O
human	B-cell_type
monocyte	I-cell_type
s	O
in	O
vitro	O
.	O

CS	O
at	O
50-200	O
microM	O
reduced	O
tumor	B-protein
necrosis	I-protein
factor	I-protein
(	O
TNF	B-protein
)	O
,	O
interleukin-1	B-protein
(	O
IL-1	B-protein
)	O
,	O
and	O
IL-6	B-protein
activities	O
in	O
the	O
culture	O
supernatants	O
from	O
LPS-stimulated	B-cell_line
human	I-cell_line
blood	I-cell_line
monocytes	I-cell_line
without	O
cytotoxicity	O
.	O

ELISA	O
assay	O
revealed	O
that	O
the	O
production	O
of	O
TNF-alpha	B-protein
,	O
IL-1alpha	B-protein
,	O
IL-1beta	B-protein
,	O
and	O
IL-6	B-protein
was	O
inhibited	O
by	O
CS	O
.	O

Northern	O
blot	O
analysis	O
showed	O
that	O
LPS-induced	O
expression	O
of	O
these	O
cytokine	B-RNA
mRNA	I-RNA
in	O
monocytes	B-cell_type
was	O
suppressed	O
by	O
CS	O
.	O

NF-kappaB	O
activation	O
was	O
also	O
inhibited	O
by	O
CS	O
.	O

These	O
findings	O
indicate	O
that	O
CS	O
has	O
a	O
suppressive	O
effect	O
on	O
proinflammatory	O
cytokine	O
production	O
from	O
monocytes	B-cell_type
,	O
and	O
this	O
effect	O
is	O
based	O
in	O
part	O
on	O
the	O
suppression	O
of	O
cytokine	B-RNA
mRNA	I-RNA
expression	O
through	O
inhibition	O
of	O
NF-kappaB	B-protein
activation	O
.	O

-DOCSTART-	O

Elevated	O
expression	O
of	O
differentiation	B-RNA
inhibitory	I-RNA
factor	I-RNA
nm23	I-RNA
mRNA	I-RNA
in	O
monoblastic	O
crisis	O
of	O
a	O
patient	O
with	O
chronic	O
myelogenous	O
leukemia	O
.	O

Differentiation	B-protein
inhibitory	I-protein
factor	I-protein
nm23	B-DNA
gene	I-DNA
has	O
been	O
found	O
to	O
be	O
expressed	O
in	O
high	O
quantities	O
in	O
acute	O
myelogenous	O
leukemia	O
(	O
AML	O
)	O
,	O
especially	O
in	O
acute	O
monocytic	O
leukemia	O
(	O
AML-M5	O
)	O
and	O
is	O
suggested	O
as	O
a	O
new	O
prognostic	O
factor	O
in	O
AML-M5	O
.	O

We	O
report	O
an	O
example	O
of	O
elevated	O
expression	O
of	O
nm23	B-RNA
mRNA	I-RNA
in	O
a	O
patient	O
with	O
chronic	O
myelogenous	O
leukemia	O
(	O
CML	O
)	O
who	O
developed	O
monoblastic	O
crisis	O
.	O

Relative	O
levels	O
of	O
nm23-H1	B-RNA
and	I-RNA
-H2	I-RNA
mRNA	I-RNA
extracted	O
from	O
the	O
patient	O
's	O
peripheral	B-cell_type
blood	I-cell_type
mononuclear	I-cell_type
cells	I-cell_type
and	O
bone	B-cell_type
marrow	I-cell_type
mononuclear	I-cell_type
cells	I-cell_type
were	O
measured	O
by	O
quantitative	O
reverse	O
transcriptase	O
polymerase	O
chain	O
reaction	O
.	O

The	O
level	O
of	O
nm23-H1	B-RNA
mRNA	I-RNA
in	O
CML	B-cell_line
cells	I-cell_line
at	O
the	O
chronic	O
phase	O
was	O
as	O
high	O
as	O
that	O
in	O
bone	B-cell_type
marrow	I-cell_type
cells	I-cell_type
from	O
healthy	O
volunteers	O
.	O

The	O
mRNA	O
level	O
of	O
nm23-H2	B-protein
was	O
slightly	O
below	O
the	O
normal	O
level	O
.	O

At	O
blastic	O
crisis	O
,	O
however	O
,	O
expression	O
of	O
both	O
nm23-H1	B-RNA
and	I-RNA
-H2	I-RNA
mRNA	I-RNA
was	O
elevated	O
to	O
about	O
three	O
to	O
nine	O
times	O
of	O
that	O
at	O
the	O
chronic	O
phase	O
.	O

Proliferated	B-cell_type
blastic	I-cell_type
cells	I-cell_type
were	O
positive	O
for	O
non-specific	B-protein
esterase	I-protein
,	O
and	O
the	O
serum	O
lysozyme	O
level	O
was	O
elevated	O
and	O
diagnosed	O
as	O
monoblastic	O
crisis	O
.	O

The	O
patient	O
received	O
combined	O
chemotherapy	O
but	O
response	O
was	O
partial	O
.	O

These	O
findings	O
are	O
compatible	O
with	O
our	O
previous	O
report	O
that	O
nm23	B-DNA
gene	I-DNA
is	O
overexpressed	O
in	O
monocytic	O
leukemia	O
.	O

-DOCSTART-	O

Increased	O
transcription	O
decreases	O
the	O
spontaneous	O
mutation	O
rate	O
at	O
the	O
thymidine	B-DNA
kinase	I-DNA
locus	I-DNA
in	O
human	B-cell_type
cells	I-cell_type
.	O

Transcription	O
increases	O
DNA	O
repair	O
efficiency	O
and	O
modulates	O
the	O
distribution	O
of	O
certain	O
types	O
of	O
DNA	O
damage	O
.	O

Furthermore	O
,	O
increased	O
transcription	O
level	O
stimulates	O
spontaneous	O
mutation	O
rate	O
in	O
yeast	O
.	O

We	O
explored	O
whether	O
transcription	O
level	O
affects	O
spontaneous	O
mutation	O
rate	O
in	O
human	B-cell_type
cells	I-cell_type
.	O

We	O
first	O
developed	O
two	O
thymidine	B-cell_line
kinase	I-cell_line
(	I-cell_line
tk	I-cell_line
)	I-cell_line
inducible	I-cell_line
human	I-cell_line
cell	I-cell_line
lines	I-cell_line
using	O
the	O
Gal4-Estrogen	O
receptor	O
system	O
.	O

In	O
our	O
TK6i-G3	B-cell_line
and	I-cell_line
G9	I-cell_line
tk	I-cell_line
heterozygous	I-cell_line
cell	I-cell_line
lines	I-cell_line
,	O
the	O
active	O
tk	B-DNA
allele	I-DNA
is	O
linked	O
to	O
an	O
inducible	O
promoter	B-DNA
element	I-DNA
.	O

Tk	B-RNA
mRNA	I-RNA
is	O
induced	O
following	O
treatment	O
with	O
estrogen	O
.	O

Spontaneous	O
mutation	O
rate	O
was	O
significantly	O
decreased	O
in	O
human	B-cell_line
cell	I-cell_line
lines	I-cell_line
after	O
induction	O
in	O
contrast	O
to	O
the	O
report	O
in	O
yeast	O
.	O

Thus	O
,	O
humans	O
may	O
have	O
evolved	O
different	O
or	O
additional	O
mechanisms	O
to	O
deal	O
with	O
transcription	O
related	O
spontaneous	O
mutagenesis	O
.	O

Copyright	O
1998	O
Elsevier	O
Science	O
B.V	O
.	O

All	O
rights	O
reserved	O
.	O

-DOCSTART-	O

Mutation	O
of	O
BCL-6	B-DNA
gene	I-DNA
in	O
normal	O
B	B-cell_type
cells	I-cell_type
by	O
the	O
process	O
of	O
somatic	O
hypermutation	O
of	O
Ig	B-DNA
genes	I-DNA
.	O

Immunoglobulin	B-DNA
(	I-DNA
Ig	I-DNA
)	I-DNA
genes	I-DNA
are	O
hypermutated	O
in	O
B	B-cell_type
lymphocytes	I-cell_type
that	O
are	O
the	O
precursors	O
to	O
memory	B-cell_type
B	I-cell_type
cells	I-cell_type
.	O

The	O
mutations	O
are	O
linked	O
to	O
transcription	O
initiation	O
,	O
but	O
non-Ig	B-DNA
promoters	I-DNA
are	O
permissible	O
for	O
the	O
mutation	O
process	O
;	O
thus	O
,	O
other	O
genes	O
expressed	O
in	O
mutating	O
B	B-cell_type
cells	I-cell_type
may	O
also	O
be	O
subject	O
to	O
somatic	O
hypermutation	O
.	O

Significant	O
mutations	O
were	O
not	O
observed	O
in	O
c-MYC	B-DNA
,	I-DNA
S14	I-DNA
,	I-DNA
or	I-DNA
alpha-fetoprotein	I-DNA
(	I-DNA
AFP	I-DNA
)	I-DNA
genes	I-DNA
,	O
but	O
BCL-6	B-protein
was	O
highly	O
mutated	O
in	O
a	O
large	O
proportion	O
of	O
memory	B-cell_type
B	I-cell_type
cells	I-cell_type
of	O
normal	O
individuals	O
.	O

The	O
mutation	O
pattern	O
was	O
similar	O
to	O
that	O
of	O
Ig	B-DNA
genes	I-DNA
.	O

-DOCSTART-	O

CD28-mediated	O
activation	O
in	O
CD45RA+	B-cell_type
and	I-cell_type
CD45RO+	I-cell_type
T	I-cell_type
cells	I-cell_type
:	O
enhanced	O
levels	O
of	O
reactive	O
oxygen	O
intermediates	O
and	O
c-Rel	O
nuclear	O
translocation	O
in	O
CD45RA+	B-cell_type
cells	I-cell_type
.	O

We	O
have	O
analyzed	O
the	O
effect	O
of	O
complete	O
T	O
cell	O
activation	O
(	O
anti-CD3	O
plus	O
anti-CD28	O
)	O
on	O
the	O
activation	O
of	O
NF-kappaB	B-protein
in	O
CD45RA+	B-cell_type
(	I-cell_type
naive	I-cell_type
)	I-cell_type
and	I-cell_type
CD45RO+	I-cell_type
(	I-cell_type
memory/effector	I-cell_type
)	I-cell_type
T	I-cell_type
cells	I-cell_type
.	O

Long	O
exposure	O
(	O
24	O
h	O
)	O
induced	O
stronger	O
NF-kappaB	B-protein
DNA	O
binding	O
in	O
CD45RA+	B-cell_type
cells	I-cell_type
than	O
in	O
CD45RO+	B-cell_type
cells	I-cell_type
.	O

Analysis	O
of	O
the	O
nuclear	B-protein
c-Rel	I-protein
protein	I-protein
indicated	O
that	O
after	O
anti-CD3+anti-CD28	O
stimulation	O
the	O
level	O
of	O
c-Rel	B-protein
was	O
higher	O
in	O
CD45RA+	B-cell_type
cells	I-cell_type
.	O

Analysis	O
of	O
the	O
cytoplasmic	B-protein
inhibitor	I-protein
IkappaBalpha	B-protein
indicated	O
that	O
anti-CD3+anti-CD28	O
stimulation	O
induced	O
a	O
long-lasting	O
degradation	O
in	O
CD45RA+	B-cell_type
cells	I-cell_type
but	O
in	O
CD45RO+	B-cell_type
cells	I-cell_type
the	O
degradation	O
process	O
was	O
more	O
rapid	O
.	O

Because	O
the	O
CD28	B-protein
costimulus	O
is	O
known	O
to	O
induce	O
the	O
production	O
of	O
reactive	O
oxygen	O
intermediates	O
(	O
ROIs	O
)	O
,	O
the	O
intracellular	O
ROI	O
levels	O
in	O
CD45RA+	B-cell_type
and	I-cell_type
CD45RO+	I-cell_type
cells	I-cell_type
were	O
compared	O
by	O
flow	O
cytometry	O
.	O

ROIs	O
were	O
produced	O
in	O
both	O
cell	O
types	O
,	O
but	O
more	O
strongly	O
in	O
CD45RA+	B-cell_type
cells	I-cell_type
.	O

The	O
data	O
presented	O
in	O
this	O
study	O
further	O
emphasize	O
the	O
differences	O
between	O
CD45RA+	B-cell_type
and	I-cell_type
CD45RO+	I-cell_type
T	I-cell_type
lymphocytes	I-cell_type
in	O
ROI-dependent	O
signaling	O
pathways	O
.	O

-DOCSTART-	O

Non-Hodgkin	O
's	O
lymphoma	O
involving	O
bilateral	O
breasts	O
[	O
see	O
comments	O
]	O

We	O
describe	O
here	O
two	O
cases	O
of	O
diffuse	O
large	O
cell	O
type	O
non-Hodgkin	O
's	O
lymphoma	O
affecting	O
the	O
bilateral	O
breasts	O
.	O

The	O
contralateral	O
tumor	O
in	O
one	O
case	O
appeared	O
17	O
months	O
after	O
the	O
first	O
mastectomy	O
,	O
whereas	O
the	O
bilateral	O
tumors	O
occurred	O
concurrently	O
in	O
the	O
other	O
patient	O
who	O
was	O
pregnant	O
and	O
showed	O
widespread	O
dissemination	O
at	O
initial	O
presentation	O
.	O

Lymphoma	B-cell_type
cells	I-cell_type
from	O
both	O
cases	O
showed	O
the	O
mature	O
B-cell	O
immunophenotype	O
and	O
had	O
rearrangements	O
of	O
the	O
BCL6	B-DNA
gene	I-DNA
.	O

Both	O
patients	O
developed	O
progressive	O
disease	O
despite	O
chemo-radiotherapy	O
and	O
died	O
of	O
leukemic	O
manifestations	O
.	O

There	O
were	O
no	O
apparent	O
pathological	O
features	O
of	O
lymphomas	O
of	O
mucosa-associated	O
lymphoid	O
tissue	O
origin	O

-DOCSTART-	O

Biochemical	O
characterization	O
of	O
MIP-1	B-protein
alpha	I-protein
nuclear	I-protein
protein	I-protein
.	O

A	O
family	O
of	O
hematopoietic	B-protein
specific	I-protein
transcription	I-protein
factors	I-protein
,	O
MIP-1	B-protein
alpha	I-protein
nuclear	I-protein
protein	I-protein
(	I-protein
MNP	I-protein
)	I-protein
family	I-protein
,	O
has	O
recently	O
been	O
identified	O
.	O

They	O
are	O
intimately	O
involved	O
in	O
regulating	O
the	O
transcription	O
of	O
the	O
huMIP-1	B-DNA
alpha	I-DNA
gene	I-DNA
in	O
monocytes	B-cell_type
,	O
T-cells	B-cell_type
,	O
and	O
transformed	B-cell_type
B-cells	I-cell_type
.	O

One	O
member	O
of	O
the	O
family	O
(	O
MNP-1	B-protein
)	O
is	O
essential	O
for	O
promoter	O
activity	O
in	O
monocytes	B-cell_type
and	O
B-cells	B-cell_type
,	O
while	O
another	O
(	O
MNP-2	B-protein
)	O
is	O
required	O
for	O
full	O
promotor	O
activity	O
in	O
T-cells	B-cell_type
.	O

A	O
third	O
member	O
of	O
the	O
family	O
(	O
MNP-3	B-protein
)	O
is	O
expressed	O
in	O
PMA	B-cell_line
induced	I-cell_line
HL60	I-cell_line
cells	I-cell_line
and	O
probably	O
has	O
a	O
role	O
in	O
monocyte	O
differentiation	O
.	O

In	O
this	O
communication	O
we	O
demonstrate	O
by	O
two	O
techniques	O
that	O
MNP-1	B-protein
and	O
MNP-2	B-protein
are	O
distinct	O
but	O
related	O
factors	O
,	O
and	O
we	O
present	O
further	O
evidence	O
to	O
show	O
that	O
MNP-1	B-protein
acts	O
as	O
a	O
heterodimer	B-protein
.	O

-DOCSTART-	O

Bcl-3	B-protein
expression	O
and	O
nuclear	O
translocation	O
are	O
induced	O
by	O
granulocyte-macrophage	B-protein
colony-stimulating	I-protein
factor	I-protein
and	O
erythropoietin	B-protein
in	O
proliferating	B-cell_type
human	I-cell_type
erythroid	I-cell_type
precursors	I-cell_type
.	O

Bcl-3	B-protein
is	O
a	O
proto-oncogene	B-DNA
involved	O
in	O
the	O
chromosomal	O
translocation	O
t	B-DNA
(	I-DNA
14	I-DNA
;	I-DNA
19	I-DNA
)	I-DNA
found	O
in	O
some	O
patients	O
with	O
chronic	O
lymphocytic	O
leukemia	O
.	O

It	O
shares	O
structural	O
similarities	O
with	O
and	O
is	O
a	O
member	O
of	O
the	O
IkappaB	B-protein
family	I-protein
of	O
proteins	O
.	O

In	O
this	O
report	O
,	O
involvement	O
of	O
Bcl-3	B-protein
in	O
hematopoietic	O
growth	O
factor-stimulated	O
erythroid	O
proliferation	O
and	O
differentiation	O
was	O
examined	O
.	O

In	O
TF-1	B-cell_line
cells	I-cell_line
,	O
an	O
erythroleukemia	B-cell_line
cell	I-cell_line
line	I-cell_line
,	O
granulocyte-macrophage	B-protein
colony-stimulating	I-protein
factor	I-protein
(	O
GM-CSF	B-protein
)	O
and	O
erythropoietin	B-protein
(	O
Epo	B-protein
)	O
greatly	O
enhanced	O
Bcl-3	B-protein
expression	O
at	O
both	O
the	O
protein	O
and	O
mRNA	O
levels	O
in	O
association	O
with	O
stimulation	O
of	O
proliferation	O
.	O

Bcl-3	B-protein
protein	O
was	O
also	O
highly	O
expressed	O
in	O
early	B-cell_type
burst-forming	I-cell_type
unit-erythroid	I-cell_type
(	I-cell_type
BFU-E	I-cell_type
)	I-cell_type
-derived	I-cell_type
erythroid	I-cell_type
precursors	I-cell_type
(	O
day	O
7	O
)	O
and	O
decreased	O
during	O
maturation	O
(	O
days	O
10	O
and	O
14	O
)	O
,	O
suggesting	O
that	O
Bcl-3	B-protein
is	O
involved	O
in	O
normal	O
erythroid	O
proliferation	O
.	O

In	O
these	O
hematopoietic	B-cell_type
cells	I-cell_type
,	O
Bcl-3	B-protein
was	O
hyperphosphorylated	O
.	O

GM-CSF	B-protein
and	O
Epo	B-protein
modulated	O
the	O
subcellular	O
localization	O
of	O
Bcl-3	B-protein
.	O

Upon	O
stimulation	O
of	O
TF-1	B-cell_line
cells	I-cell_line
with	O
GM-CSF	B-protein
or	O
Epo	B-protein
,	O
the	O
nuclear	O
translocation	O
of	O
Bcl-3	B-protein
was	O
dramatically	O
enhanced	O
.	O

Overexpression	O
of	O
Bcl-3	B-protein
in	O
TF-1	B-cell_line
cells	I-cell_line
by	O
transient	O
transfection	O
along	O
with	O
the	O
NF-kappaB	B-protein
factors	I-protein
p50	I-protein
or	I-protein
p52	I-protein
resulted	O
in	O
significant	O
induction	O
of	O
an	O
human	B-DNA
immunodeficiency	I-DNA
virus-type	I-DNA
1	I-DNA
(	I-DNA
HIV-1	I-DNA
)	I-DNA
kappaB-TATA-luceriferase	I-DNA
reporter	I-DNA
plasmid	I-DNA
,	O
demonstrating	O
that	O
Bcl-3	B-protein
has	O
a	O
positive	O
role	O
in	O
transactivation	O
of	O
kappaB-containing	B-DNA
genes	I-DNA
in	O
erythroid	B-cell_type
cells	I-cell_type
.	O

Stimulation	O
with	O
GM-CSF	B-protein
enhanced	O
c-myb	B-RNA
mRNA	I-RNA
expression	O
in	O
these	O
cells	O
.	O

Bcl-3	B-protein
in	O
nuclear	O
extracts	O
of	O
TF-1	B-cell_line
cells	I-cell_line
bound	O
to	O
a	O
kappaB	B-DNA
enhancer	I-DNA
in	O
the	O
c-myb	B-DNA
promoter	I-DNA
together	O
with	O
NF-kappaB2/p52	B-protein
and	O
this	O
binding	O
activity	O
was	O
enhanced	O
by	O
GM-CSF	B-protein
stimulation	O
.	O

Furthermore	O
,	O
cotransfection	O
of	O
Bcl-3	B-protein
with	O
p52	B-protein
or	O
p50	B-protein
in	O
TF-1	B-cell_line
cells	I-cell_line
resulted	O
in	O
significant	O
activation	O
of	O
a	O
c-myb	B-DNA
kappaB-TATA-luceriferase	I-DNA
reporter	I-DNA
plasmid	I-DNA
.	O

These	O
findings	O
suggest	O
that	O
Bcl-3	B-protein
may	O
participate	O
in	O
the	O
transcriptional	O
regulation	O
of	O
certain	O
kappaB-containing	B-DNA
genes	I-DNA
involved	O
in	O
hematopoiesis	O
,	O
including	O
c-myb	B-DNA
.	O

Copyright	O
1998	O
by	O
The	O
American	O
Society	O
of	O
Hematology	O
.	O

-DOCSTART-	O

5-Lipoxygenase	B-protein
compartmentalization	O
in	O
granulocytic	O
cells	O
is	O
modulated	O
by	O
an	O
internal	B-protein
bipartite	I-protein
nuclear	I-protein
localizing	I-protein
sequence	I-protein
and	O
nuclear	B-protein
factor	I-protein
kappa	I-protein
B	I-protein
complex	I-protein
formation	O
.	O

A	O
region	O
of	O
basic	O
amino	O
acids	O
spanning	O
residues	B-protein
639-656	I-protein
in	O
the	O
human	B-protein
5-lipoxygenase	I-protein
sequence	I-protein
resembles	O
a	O
consensus	B-protein
bipartite	I-protein
nuclear	I-protein
localizing	I-protein
sequence	I-protein
.	O

A	O
synthetic	O
peptide	O
consisting	O
of	O
the	O
Kaposi	B-protein
fibroblast	I-protein
growth	I-protein
factor	I-protein
signal	I-protein
sequence	I-protein
fused	O
to	O
the	O
5-lipoxygenase639-656	B-protein
bipartite	I-protein
nuclear	I-protein
localizing	I-protein
sequence	I-protein
has	O
a	O
prominent	O
inhibitory	O
effect	O
on	O
5-lipoxygenase	B-protein
catalysis	O
in	O
granulocytic	O
HL-60	B-cell_line
cells	I-cell_line
activated	O
by	O
calcium	O
ionophor	O
A23187	O
.	O

Recombinant	O
5-lipoxygenase	B-protein
was	O
not	O
affected	O
by	O
the	O
peptide	O
.	O

The	O
peptide	O
also	O
inhibited	O
redistribution	O
of	O
5-lipoxygenase	B-protein
from	O
the	O
cytosol	O
to	O
the	O
nuclear	O
membrane	O
of	O
HL-60	B-cell_line
cells	I-cell_line
stimulated	O
by	O
A23187	O
.	O

5-Lipoxygenase	B-protein
protein	I-protein
was	O
detected	O
in	O
nuclear	B-protein
factor	I-protein
kappaB	I-protein
(	I-protein
NF-kappaB	I-protein
)	I-protein
p65	I-protein
subunit	I-protein
immunoprecipitate	O
fractions	O
prepared	O
from	O
HL-60	B-cell_line
cell	I-cell_line
lysates	O
.	O

The	O
amount	O
of	O
5-lipoxygenase	B-protein
protein	I-protein
coimmunoprecipitated	O
by	O
NF-kappaB	O
antiserum	O
was	O
increased	O
following	O
A23187	O
stimulation	O
.	O

In	O
cells	O
treated	O
with	O
agents	O
that	O
block	O
5-lipoxygenase	B-protein
translocation	O
to	O
the	O
nucleus	O
,	O
5-lipoxygenase	B-protein
protein	O
appearing	O
in	O
the	O
NF-kappaB	B-protein
immunoprecipitate	O
was	O
diminished	O
.	O

Our	O
results	O
implicate	O
an	O
internal	B-protein
bipartite	I-protein
nuclear	I-protein
localizing	I-protein
sequence	I-protein
as	O
a	O
regulatory	O
domain	O
that	O
modulates	O
5-lipoxygenase	B-protein
redistribution	O
and	O
catalysis	O
in	O
granulocytic	B-cell_type
cells	I-cell_type
.	O

Additionally	O
,	O
our	O
results	O
suggest	O
that	O
molecular	O
determinants	O
which	O
govern	O
5-lipoxygenase	O
and	O
NF-kappaB	O
redistribution	O
to	O
the	O
nucleus	O
may	O
be	O
coordinately	O
controlled	O
in	O
granulocytic	B-cell_type
cells	I-cell_type
.	O

Copyright	O
1998	O
Academic	O
Press	O
.	O

-DOCSTART-	O

Inhibition	O
of	O
CD28/CD3-mediated	O
costimulation	O
of	O
naive	B-cell_type
and	I-cell_type
memory	I-cell_type
human	I-cell_type
T	I-cell_type
lymphocytes	I-cell_type
by	O
intracellular	O
incorporation	O
of	O
polyclonal	B-protein
antibodies	I-protein
specific	O
for	O
the	O
activator	B-protein
protein-1	I-protein
transcriptional	I-protein
complex	I-protein
.	O

A	O
number	O
of	O
indirect	O
methods	O
have	O
been	O
utilized	O
in	O
demonstrating	O
activator	B-protein
protein-1	I-protein
transcription	O
factor	O
function	O
in	O
IL-2	O
promoter	O
activity	O
.	O

However	O
,	O
there	O
has	O
been	O
no	O
direct	O
demonstration	O
that	O
activator	B-protein
protein-1	I-protein
is	O
involved	O
in	O
CD28-dependent	O
costimulation	O
of	O
IL-2	B-DNA
gene	I-DNA
transcription	O
in	O
freshly	O
isolated	O
naive	B-cell_type
and	I-cell_type
memory	I-cell_type
human	I-cell_type
T	I-cell_type
lymphocytes	I-cell_type
.	O

To	O
address	O
this	O
issue	O
,	O
the	O
method	O
of	O
scrape	O
loading	O
was	O
applied	O
to	O
purified	B-cell_type
peripheral	I-cell_type
blood	I-cell_type
T	I-cell_type
lymphocytes	I-cell_type
.	O

Since	O
scrape	O
loading	O
relies	O
on	O
adherent	B-cell_type
cells	I-cell_type
,	O
peripheral	B-cell_type
blood	I-cell_type
human	I-cell_type
T	I-cell_type
(	I-cell_type
PB-T	I-cell_type
)	I-cell_type
cells	I-cell_type
were	O
immobilized	O
on	O
the	O
nonspecific	O
cell	O
attachment	O
factor	O
poly-L-lysine	O
.	O

Cells	O
scraped	O
off	O
poly-L-lysine	O
in	O
the	O
presence	O
of	O
Ig	B-protein
FITC	I-protein
efficiently	O
incorporated	O
Ig	O
,	O
with	O
relatively	O
uniform	O
fluorescence	O
.	O

T	O
cells	O
retained	O
their	O
physical	O
parameters	O
as	O
measured	O
by	O
forward	O
and	O
side	O
light	O
scatter	O
,	O
and	O
functional	O
activity	O
as	O
measured	O
by	O
costimulation	O
of	O
proliferation	O
and	O
IL-2	B-protein
production	O
after	O
being	O
scraped	O
off	O
this	O
substrate	O
.	O

CD28/CD3-costimulated	B-cell_type
T	I-cell_type
cells	I-cell_type
produced	O
intracellular	O
IL-2	B-protein
from	O
all	O
subsets	O
measured	O
(	O
CD4+	B-cell_type
,	O
CD4-	B-cell_type
,	O
CD45RO+	B-cell_type
,	O
and	O
CD45RO-	B-cell_type
)	O
.	O

IL-2	B-protein
production	O
and	O
intracellular	O
accumulation	O
in	O
nonscraped	O
PB-T	B-cell_type
cells	I-cell_type
activated	O
with	O
CD28/CD3	O
coligation	O
were	O
skewed	O
favoring	O
CD45RO+	B-cell_type
and	I-cell_type
CD4+	I-cell_type
subsets	I-cell_type
,	O
as	O
was	O
IL-2	B-protein
production	O
in	O
scraped	O
PB-T	B-cell_type
cells	I-cell_type
.	O

The	O
intracellular	O
incorporation	O
of	O
Abs	O
specific	O
for	O
c-Fos	B-protein
and	I-protein
c-Jun	I-protein
family	I-protein
members	I-protein
by	O
scrape	O
loading	O
inhibited	O
the	O
production	O
and	O
intracellular	O
accumulation	O
of	O
IL-2	B-protein
within	O
6	O
h	O
of	O
costimulation	O
with	O
PMA/ionomycin	O
,	O
or	O
costimulation	O
by	O
CD28	O
and	O
CD3	O
ligation	O
.	O

Scrape	O
loading	O
thus	O
provides	O
an	O
efficient	O
mechanism	O
for	O
intracellular	O
incorporation	O
of	O
macromolecules	O
,	O
and	O
the	O
first	O
direct	O
evidence	O
that	O
c-Fos	B-protein
and	O
c-Jun	B-protein
are	O
involved	O
in	O
transcription	O
of	O
the	O
IL-2	B-DNA
gene	I-DNA
within	O
its	O
correct	O
chromosomal	O
context	O
,	O
in	O
resting	O
human	O
T	O
lymphocyte	O
subpopulations	O
.	O

-DOCSTART-	O

Differential	O
expression	O
of	O
Nur77	B-protein
family	I-protein
members	I-protein
in	O
human	B-cell_type
T-lymphotropic	I-cell_type
virus	I-cell_type
type	I-cell_type
1-infected	I-cell_type
cells	I-cell_type
:	O
transactivation	O
of	O
the	O
TR3/nur77	B-DNA
gene	I-DNA
by	O
Tax	B-protein
protein	I-protein
.	O

We	O
analyzed	O
the	O
differential	O
expression	O
and	O
regulation	O
of	O
three	O
members	O
of	O
the	O
Nur77	B-protein
transcription	I-protein
factor	I-protein
family	I-protein
by	O
the	O
human	O
T-lymphotropic	O
virus	O
type	O
1	O
(	O
HTLV-1	O
)	O
Tax	B-protein
protein	I-protein
.	O

We	O
have	O
demonstrated	O
that	O
in	O
both	O
HTLV-1-infected	B-cell_type
cells	I-cell_type
and	O
Tax-expressing	B-cell_line
JPX-9	I-cell_line
cells	I-cell_line
,	O
TR3/nur77	B-protein
is	O
highly	O
expressed	O
,	O
whereas	O
neither	O
NOR-1	O
nor	O
NOT	O
expression	O
is	O
detectable	O
.	O

Transient	O
transfection	O
analysis	O
further	O
confirmed	O
the	O
Tax	B-protein
transactivation	O
of	O
the	O
TR3/nur77	B-protein
promoter	O
but	O
not	O
the	O
NOR-1	B-DNA
promoter	I-DNA
in	O
different	O
cell	O
types	O
.	O

Furthermore	O
,	O
expression	O
of	O
a	O
luciferase	B-DNA
reporter	I-DNA
gene	I-DNA
driven	O
by	O
the	O
NGFI-B	B-DNA
(	I-DNA
rat	I-DNA
homolog	I-DNA
of	I-DNA
TR3/Nur77	I-DNA
)	I-DNA
response	I-DNA
element	I-DNA
(	O
NBRE	B-DNA
)	O
provided	O
evidence	O
that	O
Tax	B-protein
-mediated	O
transactivation	O
resulted	O
in	O
the	O
induction	O
of	O
a	O
functional	O
protein	O
.	O

Cotransfection	O
assays	O
with	O
the	O
TR3/nur77	B-DNA
promoter	I-DNA
sequence	I-DNA
or	O
the	O
NBRE	B-DNA
binding	O
motif	O
together	O
with	O
a	O
series	O
of	O
Tax	B-protein
mutants	O
have	O
shown	O
that	O
Tax	B-protein
-induced	O
TR3/nur77	B-protein
expression	O
is	O
mediated	O
by	O
CREB/ATF-related	B-protein
transcription	I-protein
factors	I-protein
.	O

-DOCSTART-	O

Negative	O
regulation	O
of	O
the	O
heat	O
shock	O
transcriptional	O
response	O
by	O
HSBP1	B-protein
.	O

In	O
response	O
to	O
stress	O
,	O
heat	B-protein
shock	I-protein
factor	I-protein
1	I-protein
(	O
HSF1	B-protein
)	O
acquires	O
rapid	O
DNA	O
binding	O
and	O
transient	O
transcriptional	O
activity	O
while	O
undergoing	O
conformational	O
transition	O
from	O
an	O
inert	O
non-DNA-binding	B-protein
monomer	I-protein
to	O
active	O
functional	O
trimers	O
.	O

Attenuation	O
of	O
the	O
inducible	O
transcriptional	O
response	O
occurs	O
during	O
heat	O
shock	O
or	O
upon	O
recovery	O
at	O
non-stress	O
conditions	O
and	O
involves	O
dissociation	O
of	O
the	O
HSF1	B-protein
trimer	I-protein
and	O
loss	O
of	O
activity	O
.	O

We	O
have	O
used	O
the	O
hydrophobic	B-protein
repeats	I-protein
of	O
the	O
HSF1	B-protein
trimerization	O
domain	O
in	O
the	O
yeast	O
two-hybrid	O
protein	O
interaction	O
assay	O
to	O
identify	O
heat	B-protein
shock	I-protein
factor	I-protein
binding	I-protein
protein	I-protein
1	I-protein
(	O
HSBP1	B-protein
)	O
,	O
a	O
novel	O
,	O
conserved	O
,	O
76-amino-acid	B-protein
protein	I-protein
that	O
contains	O
two	O
extended	O
arrays	O
of	O
hydrophobic	B-protein
repeats	I-protein
that	O
interact	O
with	O
the	O
HSF1	B-protein
heptad	I-protein
repeats	I-protein
.	O

HSBP1	B-protein
is	O
nuclear-localized	O
and	O
interacts	O
in	O
vivo	O
with	O
the	O
active	O
trimeric	O
state	O
of	O
HSF1	B-protein
that	O
appears	O
during	O
heat	O
shock	O
.	O

During	O
attenuation	O
of	O
HSF1	B-protein
to	O
the	O
inert	O
monomer	O
,	O
HSBP1	B-protein
associates	O
with	O
Hsp70	O
.	O

HSBP1	B-protein
negatively	O
affects	O
HSF1	B-protein
DNA-binding	O
activity	O
,	O
and	O
overexpression	O
of	O
HSBP1	B-protein
in	O
mammalian	B-cell_type
cells	I-cell_type
represses	O
the	O
transactivation	O
activity	O
of	O
HSF1	B-protein
.	O

To	O
establish	O
a	O
biological	O
role	O
for	O
HSBP1	B-protein
,	O
the	O
homologous	O
Caenorhabditis	B-protein
elegans	I-protein
protein	I-protein
was	O
overexpressed	O
in	O
body	B-cell_type
wall	I-cell_type
muscle	I-cell_type
cells	I-cell_type
and	O
was	O
shown	O
to	O
block	O
activation	O
of	O
the	O
heat	O
shock	O
response	O
from	O
a	O
heat	B-DNA
shock	I-DNA
promoter-reporter	I-DNA
construct	I-DNA
.	O

Alteration	O
in	O
the	O
level	O
of	O
HSBP1	B-protein
expression	O
in	O
C.	O
elegans	O
has	O
severe	O
effects	O
on	O
survival	O
of	O
the	O
animals	O
after	O
thermal	O
and	O
chemical	O
stress	O
,	O
consistent	O
with	O
a	O
role	O
for	O
HSBP1	B-protein
as	O
a	O
negative	O
regulator	O
of	O
the	O
heat	O
shock	O
response	O
.	O

-DOCSTART-	O

Long-range	O
transcriptional	O
regulation	O
of	O
cytokine	O
gene	O
expression	O
.	O

Most	O
studies	O
on	O
the	O
control	O
of	O
cytokine	O
gene	O
expression	O
have	O
involved	O
the	O
functional	O
analysis	O
of	O
proximal	B-DNA
promoters	I-DNA
.	O

Recent	O
work	O
has	O
identified	O
distal	B-DNA
elements	I-DNA
that	O
mediate	O
long-range	O
cytokine	O
gene	O
regulation	O
and	O
has	O
implicated	O
chromatin	O
reorganization	O
in	O
regulation	O
of	O
cytokine	B-DNA
gene	I-DNA
loci	I-DNA
.	O

These	O
studies	O
have	O
begun	O
to	O
elucidate	O
the	O
basis	O
for	O
cell-specificity	O
and	O
high-level	O
expression	O
of	O
cytokine	B-DNA
genes	I-DNA
.	O

-DOCSTART-	O

A	O
signaling	O
complex	O
of	O
Ca2+-calmodulin-dependent	B-protein
protein	I-protein
kinase	I-protein
IV	I-protein
and	O
protein	B-protein
phosphatase	I-protein
2A	I-protein
[	O
see	O
comments	O
]	O

Stimulation	O
of	O
T	B-cell_type
lymphocytes	I-cell_type
results	O
in	O
a	O
rapid	O
increase	O
in	O
intracellular	O
calcium	O
concentration	O
(	O
[	O
Ca2+	O
]	O
i	O
)	O
that	O
parallels	O
the	O
activation	O
of	O
Ca2+-calmodulin-dependent	B-protein
protein	I-protein
kinase	I-protein
IV	I-protein
(	O
CaMKIV	B-protein
)	O
,	O
a	O
nuclear	B-protein
enzyme	I-protein
that	O
can	O
phosphorylate	O
and	O
activate	O
the	O
cyclic	B-protein
adenosine	I-protein
monophosphate	I-protein
(	I-protein
cAMP	I-protein
)	I-protein
response	I-protein
element-binding	I-protein
protein	I-protein
(	O
CREB	B-protein
)	O
.	O

However	O
,	O
inactivation	O
of	O
CaMKIV	B-protein
occurs	O
despite	O
the	O
sustained	O
increase	O
in	O
[	O
Ca2+	O
]	O
i	O
that	O
is	O
required	O
for	O
T	O
cell	O
activation	O
.	O

A	O
stable	O
and	O
stoichiometric	O
complex	O
of	O
CaMKIV	B-protein
with	O
protein	B-protein
serine-threonine	I-protein
phosphatase	I-protein
2A	I-protein
(	O
PP2A	B-protein
)	O
was	O
identified	O
in	O
which	O
PP2A	B-protein
dephosphorylates	O
CaMKIV	B-protein
and	O
functions	O
as	O
a	O
negative	O
regulator	O
of	O
CaMKIV	B-protein
signaling	O
.	O

In	O
Jurkat	B-cell_line
T	I-cell_line
cells	I-cell_line
,	O
inhibition	O
of	O
PP2A	B-protein
activity	O
by	O
small	O
t	O
antigen	O
enhanced	O
activation	O
of	O
CREB	B-protein
-mediated	O
transcription	O
by	O
CaMKIV	B-protein
.	O

These	O
findings	O
reveal	O
an	O
intracellular	O
signaling	O
mechanism	O
whereby	O
a	O
protein	B-protein
serine-threonine	I-protein
kinase	I-protein
(	O
CaMKIV	B-protein
)	O
is	O
regulated	O
by	O
a	O
tightly	O
associated	O
protein	B-protein
serine-threonine	I-protein
phosphatase	I-protein
(	O
PP2A	B-protein
)	O
.	O

-DOCSTART-	O

Hypoxia	O
down-regulates	O
MCP-1	B-protein
expression	O
:	O
implications	O
for	O
macrophage	O
distribution	O
in	O
tumors	O
.	O

Monocyte	B-protein
chemoattractant	I-protein
protein	I-protein
1	I-protein
(	O
MCP-1	B-protein
)	O
is	O
likely	O
to	O
contribute	O
to	O
the	O
macrophage	B-cell_type
infiltrate	I-cell_type
in	O
human	O
ovarian	O
carcinomas	O
.	O

Although	O
MCP-1	B-protein
is	O
predominantly	O
expressed	O
by	O
the	O
tumor	O
parenchyma	O
,	O
macrophages	O
accumulate	O
at	O
highest	O
density	O
in	O
necrotic	O
regions	O
,	O
which	O
are	O
associated	O
with	O
low	O
oxygen	O
tensions	O
.	O

Tumor	B-protein
necrosis	I-protein
factor	I-protein
alpha	I-protein
(	O
TNF-alpha	B-protein
)	O
can	O
stimulate	O
MCP-1	B-protein
production	O
and	O
is	O
also	O
present	O
within	O
ovarian	O
carcinomas	O
.	O

We	O
have	O
investigated	O
the	O
effect	O
of	O
hypoxia	O
both	O
on	O
MCP-1	B-protein
expression	O
in	O
ovarian	B-cell_line
cancer	I-cell_line
cell	I-cell_line
lines	I-cell_line
and	O
monocyte	O
migration	O
.	O

Hypoxia	O
down-regulated	O
TNF-alpha-induced	B-RNA
MCP-1	I-RNA
mRNA	I-RNA
and	O
protein	O
production	O
by	O
ovarian	B-cell_type
cancer	I-cell_type
cells	I-cell_type
.	O

The	O
effect	O
was	O
mimicked	O
by	O
cobalt	O
chloride	O
and	O
desferrioxamine	O
,	O
consistent	O
with	O
a	O
specific	O
oxygen-sensing	O
mechanism	O
.	O

Unlike	O
antioxidants	O
,	O
hypoxia	O
did	O
not	O
inhibit	O
nuclear	O
factor	O
KB	O
mobilization	O
.	O

Monocyte	O
migration	O
in	O
response	O
to	O
MCP-1	B-protein
was	O
also	O
diminished	O
under	O
hypoxic	O
conditions	O
.	O

Down-regulation	O
of	O
MCP-1	B-protein
expression	O
and	O
the	O
inhibition	O
of	O
monocyte	O
migration	O
are	O
independent	O
effects	O
of	O
hypoxia	O
that	O
may	O
contribute	O
to	O
the	O
distribution	O
of	O
macrophages	B-cell_type
within	O
ovarian	O
tumors	O
.	O

-DOCSTART-	O

Activation	O
of	O
nuclear	B-protein
factor	I-protein
kappa	I-protein
B	I-protein
inflammatory	O
bowel	O
disease	O
[	O
see	O
comments	O
]	O

BACKGROUND	O
:	O
Expression	O
of	O
pro-inflammatory	B-protein
cytokines	I-protein
is	O
increased	O
in	O
the	O
intestinal	O
lamina	O
propria	O
of	O
patients	O
with	O
inflammatory	O
bowel	O
disease	O
(	O
IBD	O
)	O
.	O

Nuclear	B-protein
factor	I-protein
kappa	I-protein
B	I-protein
(	O
NF	B-protein
kappa	I-protein
B	I-protein
)	O
controls	O
transcription	O
of	O
inflammation	B-DNA
genes	I-DNA
.	O

On	O
activation	O
,	O
NF	B-protein
kappa	I-protein
B	I-protein
is	O
rapidly	O
released	O
from	O
its	O
cytoplasmic	B-protein
inhibitor	I-protein
(	O
I	B-protein
kappa	I-protein
B	I-protein
)	O
,	O
transmigrates	O
into	O
the	O
nucleus	O
,	O
and	O
binds	O
to	O
DNA	B-DNA
response	I-DNA
elements	I-DNA
in	O
gene	O
promoter	O
regions	O
.	O

AIMS	O
:	O
To	O
investigate	O
whether	O
increased	O
activation	O
of	O
NF	B-protein
kappa	I-protein
B	I-protein
is	O
important	O
in	O
IBD	O
and	O
may	O
be	O
down-regulated	O
by	O
anti-inflammatory	O
treatment	O
.	O

METHODS	O
:	O
Activation	O
of	O
NF	B-protein
kappa	I-protein
B	I-protein
was	O
determined	O
by	O
western	O
blot	O
assessment	O
and	O
electrophoretic	O
mobility	O
shift	O
assay	O
in	O
nuclear	O
extracts	O
of	O
colonic	O
biopsy	O
samples	O
as	O
well	O
as	O
lamina	B-cell_type
propria	I-cell_type
mononuclear	I-cell_type
cells	I-cell_type
.	O

RESULTS	O
:	O
Nuclear	O
levels	O
of	O
NF	B-protein
kappa	I-protein
B	I-protein
p65	B-protein
are	O
increased	O
in	O
lamina	O
propria	O
biopsy	O
specimens	O
from	O
patients	O
with	O
Crohn	O
's	O
disease	O
in	O
comparison	O
with	O
patients	O
with	O
ulcerative	O
colitis	O
and	O
controls	O
.	O

Increased	O
activation	O
of	O
NF	B-protein
kappa	I-protein
B	I-protein
was	O
detected	O
in	O
lamina	B-cell_type
propria	I-cell_type
mononuclear	I-cell_type
cells	I-cell_type
from	O
patients	O
with	O
active	O
IBD	O
.	O

Corticosteroids	O
strongly	O
inhibit	O
intestinal	O
NF	B-protein
kappa	I-protein
B	I-protein
activation	O
in	O
IBD	O
in	O
vivo	O
and	O
in	O
vitro	O
by	O
stabilising	O
the	O
cytosolic	B-protein
inhibitor	I-protein
I	B-protein
kappa	I-protein
B	I-protein
alpha	O
against	O
activation	O
induced	O
degradation	O
.	O

CONCLUSIONS	O
:	O
In	O
both	O
IBDs	O
,	O
but	O
particularly	O
Crohn	O
's	O
disease	O
,	O
increased	O
activation	O
of	O
NF	B-protein
kappa	I-protein
B	I-protein
may	O
be	O
involved	O
in	O
the	O
regulation	O
of	O
the	O
inflammatory	O
response	O
.	O

Inhibition	O
of	O
NF	B-protein
kappa	I-protein
B	I-protein
activation	O
may	O
represent	O
a	O
mechanism	O
by	O
which	O
steroids	O
exert	O
an	O
anti-inflammatory	O
effect	O
in	O
IBD	O

-DOCSTART-	O

Insufficient	O
glycemic	O
control	O
increases	O
nuclear	B-protein
factor-kappa	I-protein
B	I-protein
binding	O
activity	O
in	O
peripheral	B-cell_type
blood	I-cell_type
mononuclear	I-cell_type
cells	I-cell_type
isolated	O
from	O
patients	O
with	O
type	O
1	O
diabetes	O
.	O

OBJECTIVE	O
:	O
The	O
redox-sensitive	B-protein
transcription	I-protein
factor	I-protein
nuclear	I-protein
factor-kappa	I-protein
B	I-protein
(	O
NF-kappa	B-protein
B	I-protein
)	O
is	O
believed	O
to	O
contribute	O
to	O
late	O
diabetic	O
complications	O
.	O

It	O
is	O
unknown	O
whether	O
NF-kappa	B-protein
B	I-protein
is	O
influenced	O
by	O
glycemic	O
control	O
.	O

RESEARCH	O
DESIGN	O
AND	O
METHODS	O
:	O
To	O
determine	O
whether	O
NF-kappa	B-protein
B	I-protein
is	O
activated	O
in	O
patients	O
with	O
insufficient	O
glycemic	O
control	O
(	O
HbA1c	O
>	O
10	O
%	O
)	O
,	O
we	O
developed	O
a	O
tissue	O
culture-independent	O
electrophoretic	O
mobility	O
shift	O
assay	O
(	O
EMSA	O
)	O
-based	O
semiquantitative	O
detection	O
system	O
that	O
allowed	O
us	O
to	O
determine	O
NF-kappa	B-protein
B	I-protein
activation	O
in	O
ex	O
vivo-isolated	O
peripheral	B-cell_type
blood	I-cell_type
mononuclear	I-cell_type
cells	I-cell_type
(	O
PBMCs	B-cell_type
)	O
.	O

We	O
included	O
43	O
patients	O
with	O
type	O
1	O
diabetes	O
in	O
this	O
cross-sectional	O
study	O
.	O

10	O
of	O
those	O
received	O
the	O
antioxidant	O
thioctic	O
acid	O
(	O
600	O
mg/day	O
p.o.	O
)	O
for	O
2	O
weeks	O
.	O

RESULTS	O
:	O
Monocytes	O
of	O
patients	O
with	O
HbA1c	O
levels	O
>	O
10	O
%	O
demonstrated	O
significantly	O
higher	O
NF-kappa	B-protein
B	I-protein
binding	O
activity	O
in	O
an	O
EMSA	O
and	O
a	O
stronger	O
NF-kappa	B-protein
B	I-protein
staining	O
in	O
immunohistochemistry	O
than	O
monocytes	B-cell_type
of	O
patients	O
with	O
HbA1c	O
levels	O
of	O
6-8	O
%	O
.	O

The	O
increase	O
in	O
NF-kappa	B-protein
B	I-protein
activation	O
correlated	O
with	O
an	O
increase	O
in	O
plasmatic	O
markers	O
of	O
lipid	O
peroxidation	O
.	O

Treatment	O
with	O
the	O
antioxidant	O
thioctic	O
acid	O
decreased	O
NF-kappa	B-protein
B	I-protein
binding	O
activity	O
.	O

CONCLUSIONS	O
:	O
Hyperglycemia	O
induces	O
activation	O
of	O
the	O
transcription	O
factor	O
NF-kappa	B-protein
B	I-protein
in	O
ex	O
vivo-isolated	O
PBMCs	B-cell_type
of	O
patients	O
with	O
type	O
1	O
diabetes	O
.	O

NF-kappa	B-protein
B	I-protein
activation	O
is	O
at	O
least	O
partially	O
dependent	O
on	O
oxidative	O
stress	O
,	O
since	O
the	O
antioxidant	O
thioctic	O
acid	O
significantly	O
lowered	O
the	O
extent	O
of	O
NF-kappa	B-protein
B	I-protein
binding	O
activity	O
.	O

-DOCSTART-	O

Synergistic	O
activation	O
of	O
MAP	B-protein
kinase	I-protein
(	O
ERK1/2	B-protein
)	O
by	O
erythropoietin	B-protein
and	O
stem	B-protein
cell	I-protein
factor	I-protein
is	O
essential	O
for	O
expanded	O
erythropoiesis	O
.	O

Stem	B-protein
cell	I-protein
factor	I-protein
(	O
SCF	B-protein
)	O
and	O
erythropoietin	B-protein
(	O
EPO	B-protein
)	O
work	O
synergistically	O
to	O
support	O
erythropoiesis	O
,	O
but	O
the	O
mechanism	O
for	O
this	O
synergism	O
is	O
unknown	O
.	O

By	O
using	O
purified	B-cell_line
human	I-cell_line
erythroid	I-cell_line
colony-forming	I-cell_line
cells	I-cell_line
(	O
ECFC	B-cell_line
)	O
,	O
we	O
have	O
found	O
that	O
SCF	B-protein
and	O
EPO	B-protein
synergistically	O
activate	O
MAP	B-protein
kinase	I-protein
(	O
MAPK	B-protein
,	O
ERK1/2	B-protein
)	O
,	O
which	O
correlates	O
with	O
the	O
cell	O
growth	O
and	O
thus	O
may	O
be	O
responsible	O
for	O
the	O
synergistic	O
effects	O
.	O

Treatment	O
of	O
the	O
cells	O
with	O
PD98059	O
and	O
wortmannin	O
,	O
inhibitors	O
of	O
MEK	B-protein
and	O
PI-3	B-protein
kinase	I-protein
,	O
respectively	O
,	O
inhibited	O
the	O
synergistic	O
activation	O
of	O
MAPK	B-protein
and	O
also	O
the	O
cell	O
growth	O
,	O
further	O
supporting	O
this	O
conclusion	O
.	O

Wortmannin	O
only	O
inhibits	O
MAPK	B-protein
activation	O
induced	O
by	O
EPO	B-protein
but	O
not	O
that	O
by	O
SCF	B-protein
,	O
suggesting	O
that	O
SCF	B-protein
and	O
EPO	B-protein
may	O
activate	O
MAPK	B-protein
through	O
different	O
pathways	O
,	O
which	O
would	O
facilitate	O
synergy	O
.	O

Furthermore	O
,	O
EPO	B-protein
,	O
but	O
not	O
SCF	B-protein
,	O
led	O
to	O
activation	O
of	O
STAT5	B-protein
,	O
whereas	O
SCF	B-protein
and	O
wortmannin	O
had	O
no	O
effect	O
on	O
the	O
EPO	B-protein
-induced	O
STAT5	B-protein
activation	O
,	O
suggesting	O
that	O
STAT5	B-protein
is	O
not	O
involved	O
in	O
the	O
synergistic	O
action	O
of	O
SCF	B-protein
and	O
EPO	B-protein
.	O

Together	O
,	O
the	O
data	O
suggest	O
that	O
synergistic	O
activation	O
of	O
MAPK	B-protein
by	O
SCF	B-protein
and	O
EPO	B-protein
is	O
essential	O
for	O
expanded	O
erythropoiesis	O
.	O

Copyright	O
1998	O
by	O
The	O
American	O
Society	O
of	O
Hematology	O
.	O

-DOCSTART-	O

The	O
molecular	O
and	O
phenotypic	O
profile	O
of	O
primary	O
central	O
nervous	O
system	O
lymphoma	O
identifies	O
distinct	O
categories	O
of	O
the	O
disease	O
and	O
is	O
consistent	O
with	O
histogenetic	O
derivation	O
from	O
germinal	B-cell_type
center-related	I-cell_type
B	I-cell_type
cells	I-cell_type
.	O

Primary	O
central	O
nervous	O
system	O
lymphoma	O
(	O
PCNSL	O
)	O
is	O
a	O
major	O
cause	O
of	O
morbidity	O
and	O
mortality	O
among	O
human	O
immunodeficiency	O
virus	O
(	O
HIV	O
)	O
-infected	O
individuals	O
.	O

The	O
precise	O
histogenetic	O
derivation	O
and	O
the	O
molecular	O
pathogenesis	O
of	O
PCNSL	O
is	O
poorly	O
understood	O
.	O

In	O
an	O
attempt	O
to	O
clarify	O
the	O
histogenesis	O
and	O
pathogenesis	O
of	O
these	O
lymphomas	O
,	O
49	O
PCNSL	O
(	O
26	O
acquired	O
immunodeficiency	O
syndrome	O
[	O
AIDS	O
]	O
-related	O
and	O
23	O
AIDS-unrelated	O
)	O
were	O
analyzed	O
for	O
multiple	O
biologic	O
markers	O
,	O
which	O
are	O
known	O
to	O
bear	O
histogenetic	O
and	O
pathogenetic	O
significance	O
for	O
mature	O
B-cell	O
neoplasms	O
.	O

PCNSL	O
associated	O
frequently	O
(	O
50.0	O
%	O
)	O
with	O
mutations	O
of	O
BCL-6	B-DNA
5	I-DNA
'	I-DNA
noncoding	I-DNA
regions	I-DNA
,	O
which	O
are	O
regarded	O
as	O
a	O
marker	O
of	O
B-cell	O
transition	O
through	O
the	O
germinal	O
center	O
(	O
GC	O
)	O
.	O

Expression	O
of	O
BCL-6	B-protein
protein	I-protein
,	O
which	O
is	O
restricted	O
to	O
GC	B-cell_type
B	I-cell_type
cells	I-cell_type
throughout	O
physiologic	O
B-cell	O
maturation	O
,	O
was	O
detected	O
in	O
100	O
%	O
AIDS-unrelated	O
PCNSL	O
and	O
in	O
56.2	O
%	O
AIDS-related	O
cases	O
.	O

Notably	O
,	O
among	O
AIDS-related	O
PCNSL	O
,	O
expression	O
of	O
BCL-6	B-protein
was	O
mutually	O
exclusive	O
with	O
expression	O
of	O
Epstein-Barr	B-protein
virus	I-protein
(	I-protein
EBV	I-protein
)	I-protein
-encoded	I-protein
latent	I-protein
membrane	I-protein
protein	I-protein
(	I-protein
LMP	I-protein
)	I-protein
-1	I-protein
and	O
,	O
with	O
few	O
exceptions	O
,	O
also	O
of	O
BCL-2	B-protein
.	O

All	O
but	O
one	O
PCNSL	O
expressed	O
hMSH2	O
,	O
which	O
among	O
mature	B-cell_type
B	I-cell_type
cells	I-cell_type
selectively	O
stains	O
GC	B-cell_type
B	I-cell_type
cells	I-cell_type
.	O

These	O
data	O
suggest	O
that	O
PCNSL	O
may	O
be	O
frequently	O
related	O
to	O
GC	B-cell_type
B	I-cell_type
cells	I-cell_type
and	O
may	O
be	O
segregated	O
into	O
two	O
major	O
biologic	O
categories	O
based	O
on	O
the	O
expression	O
pattern	O
of	O
BCL-6	B-protein
,	O
LMP-1	B-protein
,	O
and	O
BCL-2	B-protein
.	O

BCL-6	B-protein
(	O
+	O
)	O
/	O
LMP-1	B-protein
(	O
-	O
)	O
/	O
BCL-2	B-protein
(	O
-	O
)	O
PCNSL	O
occur	O
both	O
in	O
the	O
presence	O
and	O
in	O
the	O
absence	O
of	O
HIV	O
infection	O
and	O
consistently	O
display	O
a	O
large	O
noncleaved	O
cell	O
morphology	O
.	O

Conversely	O
,	O
BCL-6	B-protein
(	O
-	O
)	O
/	O
LMP-1	B-protein
(	O
+	O
)	O
/	O
BCL-2	B-protein
(	O
+	O
)	O
PCNSL	O
are	O
restricted	O
to	O
HIV-infected	O
hosts	O
and	O
are	O
represented	O
by	O
lymphomas	O
with	O
immunoblastic	O
features	O
.	O

These	O
data	O
are	O
relevant	O
for	O
the	O
pathogenesis	O
and	O
histogenesis	O
of	O
PCNSL	O
and	O
may	O
be	O
helpful	O
to	O
segregate	O
distinct	O
biologic	O
and	O
prognostic	O
categories	O
of	O
these	O
lymphomas	O
.	O

Copyright	O
1998	O
by	O
The	O
American	O
Society	O
of	O
Hematology	O
.	O

-DOCSTART-	O

Antioxidant	O
regulation	O
of	O
phorbol	O
ester-induced	O
adhesion	O
of	O
human	B-cell_line
Jurkat	I-cell_line
T-cells	I-cell_line
to	O
endothelial	B-cell_type
cells	I-cell_type
.	O

Regulation	O
of	O
adhesion	O
molecule	O
expression	O
and	O
function	O
by	O
reactive	O
oxygen	O
species	O
via	O
specific	O
redox	O
sensitive	O
mechanisms	O
have	O
been	O
reported	O
.	O

The	O
effects	O
of	O
clinically	O
safe	O
antioxidants	O
in	O
the	O
regulation	O
of	O
adhesion	O
molecule	O
expression	O
in	O
human	B-cell_type
endothelial	I-cell_type
cells	I-cell_type
(	O
ECV	O
)	O
,	O
and	O
adherence	O
of	O
human	B-cell_line
Jurkat	I-cell_line
T	I-cell_line
cells	I-cell_line
to	O
ECV	B-cell_type
cells	I-cell_type
were	O
investigated	O
.	O

The	O
thiol	O
antioxidant	O
,	O
alpha-lipoate	O
,	O
at	O
clinically	O
relevant	O
doses	O
down-regulated	O
phorbol	O
12-myristate	O
13-acetate	O
(	O
PMA	O
)	O
-induced	O
adhesion	O
molecule	O
expression	O
and	O
cell-cell	O
adhesion	O
.	O

Inhibition	O
of	O
PMA-induced	O
ICAM-1	O
and	O
VCAM-1	O
expression	O
as	O
well	O
as	O
PMA-induced	O
adhesion	O
of	O
Jurkat	B-cell_line
T-cells	I-cell_line
to	O
ECV	B-cell_type
cells	I-cell_type
by	O
alpha-lipoate	O
was	O
dose	O
dependent	O
(	O
50-250	O
microM	O
)	O
.	O

The	O
effect	O
was	O
significant	O
for	O
ICAM-1	B-protein
(	O
p	O
<	O
.01	O
)	O
and	O
VCAM-1	B-protein
(	O
p	O
<	O
.01	O
)	O
expression	O
in	O
cells	O
pretreated	O
with	O
100	O
microM	O
alpha-lipoate	O
compared	O
to	O
PMA-activated	B-cell_type
untreated	I-cell_type
cells	I-cell_type
.	O

Inhibition	O
of	O
PMA-induced	O
adhesion	O
molecule	O
expression	O
and	O
cell-cell	O
adhesion	O
was	O
more	O
pronounced	O
when	O
a	O
combination	O
of	O
antioxidants	O
,	O
alpha-lipoate	O
and	O
alpha-tocopherol	O
,	O
were	O
used	O
compared	O
to	O
the	O
use	O
of	O
either	O
of	O
these	O
antioxidant	O
alone	O
.	O

The	O
regulation	O
of	O
adhesion	O
molecule	O
expression	O
and	O
function	O
by	O
low	O
concentration	O
of	O
antioxidants	O
investigated	O
does	O
not	O
appear	O
to	O
be	O
NF-kappaB	B-protein
regulated	O
or	O
transcription	O
dependent	O
because	O
no	O
change	O
in	O
the	O
mRNA	O
response	O
was	O
observed	O
.	O

Protein	B-protein
kinase	I-protein
C	I-protein
(	O
PKC	B-protein
)	O
has	O
been	O
suggested	O
to	O
regulate	O
PMA-induced	O
adhesion	O
molecule	O
expression	O
by	O
post-transcriptional	O
stabilization	O
of	O
adhesion	B-RNA
molecule	I-RNA
mRNA	I-RNA
.	O

Alpha-lipoate	O
pretreatment	O
did	O
not	O
influence	O
the	O
response	O
of	O
PKC	B-protein
activity	O
to	O
PMA	O
.	O

Oxidants	O
are	O
known	O
to	O
be	O
involved	O
in	O
the	O
regulation	O
of	O
cell	O
adhesion	O
processes	O
.	O

Treatment	O
of	O
ECV	B-cell_type
cells	I-cell_type
with	O
PMA	O
induced	O
generation	O
of	O
intracellular	O
oxidants	O
.	O

Alpha-lipoate	O
(	O
100	O
or	O
250	O
microM	O
)	O
treatment	O
decreased	O
PMA-induced	O
generation	O
of	O
intracellular	O
oxidants	O
.	O

The	O
inhibitory	O
effect	O
of	O
low	O
concentration	O
of	O
alpha-lipaote	O
alone	O
or	O
in	O
combination	O
with	O
alpha-tocopherol	O
on	O
agonist-induced	O
adhesion	O
processes	O
observed	O
in	O
this	O
study	O
may	O
be	O
of	O
potential	O
therapeutic	O
value	O
.	O

-DOCSTART-	O

Binding	O
of	O
human	O
immunodeficiency	O
virus	O
type	O
1	O
to	O
CD4	B-protein
and	I-protein
CXCR4	I-protein
receptors	I-protein
differentially	O
regulates	O
expression	O
of	O
inflammatory	B-DNA
genes	I-DNA
and	O
activates	O
the	O
MEK	B-protein
/ERK	O
signaling	O
pathway	O
.	O

We	O
have	O
previously	O
shown	O
that	O
binding	O
of	O
human	O
immunodeficiency	O
virus	O
type	O
1	O
(	O
HIV-1	O
)	O
virions	O
to	O
CD4	B-protein
receptors	I-protein
stimulates	O
association	O
of	O
Lck	B-protein
with	O
Raf-1	B-protein
and	O
results	O
in	O
the	O
activation	O
of	O
Raf-1	B-protein
kinase	O
in	O
a	O
Ras-independent	O
manner	O
.	O

In	O
the	O
present	O
study	O
,	O
we	O
demonstrate	O
that	O
HIV-1	B-protein
envelope	I-protein
glycoproteins	I-protein
of	O
both	O
T-cell-tropic	O
and	O
macrophagetropic	O
strains	O
rapidly	O
activate	O
the	O
ERK/mitogen-activated	O
protein	O
(	O
MAP	O
)	O
kinase	O
pathway	O
and	O
the	O
binding	O
of	O
nuclear	B-protein
transcription	I-protein
factors	I-protein
(	O
AP-1	B-protein
,	O
NF-kappaB	B-protein
,	O
and	O
C/EBP	B-protein
)	O
and	O
stimulate	O
expression	O
of	O
cytokine	B-DNA
and	I-DNA
chemokine	I-DNA
genes	I-DNA
.	O

The	O
activation	O
of	O
this	O
signaling	O
pathway	O
requires	O
functional	O
CD4	B-protein
receptors	I-protein
and	O
is	O
independent	O
of	O
binding	O
to	O
CXCR4	B-protein
.	O

Binding	O
of	O
the	O
natural	O
ligand	O
stromal	B-protein
cell-derived	I-protein
factor	I-protein
1	I-protein
(	O
SDF-1	B-protein
)	O
to	O
CXCR4	B-protein
,	O
which	O
inhibits	O
entry	O
of	O
T-cell-tropic	O
HIV-1	O
,	O
activates	O
also	O
the	O
ERK/	O
MAP	B-protein
kinase	I-protein
pathway	O
.	O

However	O
,	O
SDF-1	B-protein
did	O
not	O
affect	O
the	O
CD4-mediated	O
expression	O
of	O
cytokine	B-DNA
and	I-DNA
chemokine	I-DNA
genes	I-DNA
.	O

These	O
results	O
provide	O
firm	O
molecular	O
evidence	O
that	O
binding	O
of	O
HIV-1	B-protein
envelope	I-protein
glycoproteins	I-protein
to	O
CD4	B-protein
receptor	I-protein
initiates	O
a	O
signaling	O
pathway	O
(	O
s	O
)	O
independent	O
of	O
the	O
binding	O
to	O
the	O
chemokine	B-protein
receptor	I-protein
that	O
leads	O
to	O
the	O
aberrant	O
expression	O
of	O
inflammatory	B-DNA
genes	I-DNA
and	O
may	O
contribute	O
significantly	O
to	O
HIV-1	O
replication	O
as	O
well	O
as	O
to	O
deregulation	O
of	O
the	O
immune	O
system	O
.	O

-DOCSTART-	O

Antigen	O
receptor	O
signaling	O
induces	O
MAP	O
kinase-mediated	O
phosphorylation	O
and	O
degradation	O
of	O
the	O
BCL-6	B-protein
transcription	B-protein
factor	I-protein
.	O

The	O
bcl-6	B-DNA
proto-oncogene	I-DNA
encodes	O
a	O
POZ/zinc	B-protein
finger	I-protein
transcriptional	I-protein
repressor	I-protein
expressed	O
in	O
germinal	B-cell_type
center	I-cell_type
(	I-cell_type
GC	I-cell_type
)	I-cell_type
B	I-cell_type
and	I-cell_type
T	I-cell_type
cells	I-cell_type
and	O
required	O
for	O
GC	O
formation	O
and	O
antibody	O
affinity	O
maturation	O
.	O

Deregulation	O
of	O
bcl-6	B-DNA
expression	O
by	O
chromosomal	O
rearrangements	O
and	O
point	O
mutations	O
of	O
the	O
bcl-6	B-DNA
promoter	I-DNA
region	I-DNA
are	O
implicated	O
in	O
the	O
pathogenesis	O
of	O
B-cell	O
lymphoma	O
.	O

The	O
signals	O
regulating	O
bcl-6	B-DNA
expression	O
are	O
not	O
known	O
.	O

Here	O
we	O
show	O
that	O
antigen	O
receptor	O
activation	O
leads	O
to	O
BCL-6	B-protein
phosphorylation	O
by	O
mitogen-activated	B-protein
protein	I-protein
kinase	I-protein
(	O
MAPK	B-protein
)	O
.	O

Phosphorylation	O
,	O
in	O
turn	O
,	O
targets	O
BCL-6	B-protein
for	O
rapid	O
degradation	O
by	O
the	O
ubiquitin	B-protein
/	O
proteasome	B-protein
pathway	O
.	O

These	O
findings	O
indicate	O
that	O
BCL-6	B-protein
expression	O
is	O
directly	O
controlled	O
by	O
the	O
antigen	O
receptor	O
via	O
MAPK	B-protein
activation	O
.	O

This	O
signaling	O
pathway	O
may	O
be	O
crucial	O
for	O
the	O
control	O
of	O
B-cell	O
differentiation	O
and	O
antibody	O
response	O
and	O
has	O
implications	O
for	O
the	O
regulation	O
of	O
other	O
POZ/zinc	B-protein
finger	I-protein
transcription	I-protein
factors	I-protein
in	O
other	O
tissues	O
.	O

-DOCSTART-	O

Glucocorticoid	B-protein
receptors	I-protein
are	O
differentially	O
expressed	O
in	O
the	O
cells	O
and	O
tissues	O
of	O
the	O
immune	O
system	O
.	O

Cytosolic	B-protein
glucocorticoid	I-protein
receptor	I-protein
(	O
GR	B-protein
)	O
binding	O
studies	O
on	O
immune	O
tissues	O
demonstrate	O
that	O
the	O
thymus	O
exhibits	O
three	O
to	O
four	O
times	O
higher	O
levels	O
of	O
GR	B-protein
protein	I-protein
than	O
the	O
spleen	O
.	O

High	O
levels	O
of	O
GR	B-protein
are	O
consistent	O
with	O
the	O
exquisite	O
sensitivity	O
of	O
the	O
thymus	O
to	O
glucocorticoid	O
exposure	O
.	O

Nevertheless	O
,	O
whole	O
cell	O
binding	O
studies	O
reveal	O
similar	O
levels	O
of	O
GR	B-protein
in	O
immature	B-cell_type
thymic	I-cell_type
T	I-cell_type
lymphocytes	I-cell_type
and	O
more	O
mature	O
,	O
splenic	B-cell_type
T	I-cell_type
lymphocytes	I-cell_type
.	O

Moreover	O
,	O
whole	O
cell	O
binding	O
techniques	O
indicate	O
that	O
neutrophils	B-cell_type
(	O
which	O
represent	O
roughly	O
30	O
%	O
of	O
splenic	B-cell_type
leukocytes	I-cell_type
)	O
exhibit	O
higher	O
GR	B-protein
than	O
both	O
T	B-cell_type
and	I-cell_type
B	I-cell_type
lymphocytes	I-cell_type
,	O
further	O
contradicting	O
results	O
from	O
cytosolic	O
binding	O
studies	O
.	O

To	O
address	O
these	O
inconsistencies	O
,	O
GR	B-protein
protein	I-protein
was	O
assessed	O
in	O
immune	B-cell_type
cells	I-cell_type
and	I-cell_type
tissues	I-cell_type
using	O
cytosolic	O
radioligand	O
binding	O
.	O

Western	O
blot	O
analysis	O
,	O
and	O
immunocytochemistry	O
.	O

Consistent	O
with	O
previous	O
cytosolic	O
receptor	O
binding	O
studies	O
on	O
immune	O
tissue	O
homogenates	O
,	O
thymic	O
T	B-cell_type
cells	I-cell_type
were	O
found	O
to	O
have	O
higher	O
levels	O
of	O
GR	B-protein
than	O
T	B-cell_type
cells	I-cell_type
isolated	O
from	O
the	O
spleen	O
.	O

In	O
addition	O
,	O
neutrophils	O
were	O
found	O
to	O
have	O
fewer	O
GR	B-protein
than	O
lymphocytes	B-cell_type
and	O
monocytes	B-cell_type
.	O

These	O
results	O
indicate	O
a	O
meaningful	O
relationship	O
between	O
receptor	O
expression	O
and	O
known	O
sensitivity	O
to	O
glucocorticoids	O
.	O

-DOCSTART-	O

Functional	O
replacement	O
of	O
the	O
mouse	B-DNA
E2A	I-DNA
gene	I-DNA
with	O
a	O
human	B-DNA
HEB	I-DNA
cDNA	I-DNA
.	O

The	O
mammalian	B-DNA
E2A	I-DNA
,	I-DNA
HEB	I-DNA
,	I-DNA
and	I-DNA
E2-2	I-DNA
genes	I-DNA
encode	O
a	O
unique	O
class	O
of	O
basic	B-protein
helix-loop-helix	I-protein
(	I-protein
bHLH	I-protein
)	I-protein
transcription	I-protein
factors	I-protein
that	O
are	O
evolutionarily	O
conserved	O
and	O
essential	O
for	O
embryonic	O
and	O
postnatal	O
development	O
.	O

While	O
the	O
structural	O
and	O
functional	O
similarities	O
among	O
the	O
gene	B-protein
products	I-protein
are	O
well	O
demonstrated	O
,	O
it	O
is	O
not	O
clear	O
why	O
deletion	O
of	O
E2A	B-DNA
,	O
but	O
not	O
HEB	B-DNA
or	O
E2-2	B-DNA
,	O
leads	O
to	O
a	O
complete	O
arrest	O
in	O
B-lymphocyte	O
development	O
.	O

To	O
understand	O
the	O
molecular	O
basis	O
of	O
the	O
functional	O
specificity	O
between	O
E2A	B-DNA
and	O
HEB	B-DNA
/	O
E2-2	B-DNA
in	O
mammalian	O
development	O
,	O
we	O
generated	O
and	O
tested	O
a	O
panel	O
of	O
E2A	B-DNA
knockin	O
mutations	O
including	O
subtle	O
mutations	O
in	O
the	O
E12	B-DNA
and	I-DNA
E47	I-DNA
exons	I-DNA
and	O
substitution	O
of	O
both	O
E12	B-DNA
and	I-DNA
E47	I-DNA
exons	I-DNA
with	O
a	O
human	B-DNA
HEB	I-DNA
cDNA	I-DNA
.	O

We	O
find	O
that	O
the	O
alternatively	O
spliced	O
E12	B-protein
and	I-protein
E47	I-protein
bHLH	I-protein
proteins	I-protein
of	O
the	O
E2A	B-DNA
gene	I-DNA
play	O
similar	O
and	O
additive	O
roles	O
in	O
supporting	O
B	B-cell_type
lymphopoiesis	I-cell_type
.	O

Further	O
,	O
we	O
find	O
that	O
HEB	B-DNA
driven	O
by	O
the	O
endogenous	B-DNA
E2A	I-DNA
promoter	I-DNA
can	O
functionally	O
replace	O
E2A	B-DNA
in	O
supporting	O
B-cell	O
commitment	O
and	O
differentiation	O
toward	O
completion	O
.	O

Finally	O
,	O
the	O
postnatal	O
lethality	O
associated	O
with	O
E2A	B-DNA
disruption	O
is	O
fully	O
rescued	O
by	O
the	O
addition	O
of	O
HEB	B-DNA
.	O

This	O
study	O
suggests	O
that	O
the	O
functional	O
divergence	O
among	O
E12	B-DNA
,	O
E47	B-DNA
,	O
and	O
HEB	B-DNA
in	O
different	O
cell	O
types	O
is	O
partially	O
defined	O
by	O
the	O
context	O
of	O
gene	O
expression	O
.	O

-DOCSTART-	O

Biased	O
dependency	O
of	O
CD80	B-protein
versus	O
CD86	B-protein
in	O
the	O
induction	O
of	O
transcription	B-protein
factors	I-protein
regulating	O
the	O
human	B-DNA
IL-2	I-DNA
promoter	I-DNA
.	O

In	O
addition	O
to	O
the	O
signals	O
obtained	O
by	O
ligation	O
of	O
the	O
TCR	O
,	O
T	B-cell_type
cells	I-cell_type
need	O
additional	O
,	O
co-stimulatory	O
signals	O
to	O
be	O
activated	O
.	O

One	O
such	O
co-stimulatory	O
signal	O
is	O
delivered	O
when	O
CD28	B-protein
on	O
T	B-cell_type
cells	I-cell_type
binds	O
to	O
CD80	B-protein
or	O
CD86	B-protein
on	O
antigen-presenting	B-cell_type
cells	I-cell_type
(	O
APC	B-cell_type
)	O
.	O

In	O
the	O
present	O
study	O
,	O
we	O
analyzed	O
the	O
ability	O
of	O
CD80	B-protein
and	O
CD86	B-protein
to	O
co-stimulate	B-cell_type
human	I-cell_type
T	I-cell_type
cells	I-cell_type
activated	O
by	O
superantigen	O
.	O

Using	O
the	O
Raji	B-cell_line
B	I-cell_line
cell	I-cell_line
lymphoma	I-cell_line
,	O
which	O
express	O
similar	O
levels	O
of	O
CD80	B-protein
and	O
CD86	B-protein
,	O
it	O
was	O
found	O
that	O
T	O
cell	O
proliferation	O
was	O
mainly	O
co-stimulated	O
by	O
CD80	B-protein
.	O

To	O
further	O
characterize	O
the	O
consequences	O
of	O
this	O
biased	O
co-stimulatory	O
dependency	O
,	O
we	O
employed	O
a	O
well-defined	O
system	O
of	O
transfected	O
CHO	B-cell_line
cells	I-cell_line
expressing	O
human	B-protein
MHC	I-protein
class	I-protein
II	I-protein
together	O
with	O
CD80	B-protein
,	O
CD86	B-protein
or	O
CD80	B-protein
and	O
CD86	B-protein
.	O

Proliferation	O
of	O
freshly	O
prepared	O
CD4+	B-cell_type
T	I-cell_type
cells	I-cell_type
required	O
the	O
presence	O
of	O
either	O
CD80	B-protein
or	O
CD86	B-protein
.	O

However	O
,	O
IL-2	B-protein
production	O
reached	O
only	O
suboptimal	O
levels	O
in	O
the	O
presence	O
of	O
CD86	B-protein
but	O
optimal	O
levels	O
with	O
CD80	B-protein
.	O

To	O
analyze	O
IL-2	B-protein
transcriptional	O
activity	O
in	O
CD80	B-protein
and	O
CD86	B-protein
co-stimulated	O
T	B-cell_type
cells	I-cell_type
we	O
used	O
Jurkat	B-cell_line
T	I-cell_line
cells	I-cell_line
transfected	O
with	O
luciferase	B-DNA
reporter	I-DNA
gene	I-DNA
constructs	I-DNA
.	O

CD80	B-protein
induced	O
higher	O
levels	O
of	O
IL-2	B-DNA
promoter	I-DNA
-enhancer	O
activity	O
compared	O
to	O
CD86	B-protein
.	O

Furthermore	O
,	O
the	O
activity	O
of	O
transcription	B-protein
factors	I-protein
regulating	O
the	O
IL-2	B-DNA
promoter-enhancer	I-DNA
region	I-DNA
including	O
activation	B-protein
protein-1	I-protein
,	O
CD28	B-DNA
response	I-DNA
element	I-DNA
and	O
nuclear	B-protein
factor	I-protein
kappaB	I-protein
were	O
4-8	O
times	O
higher	O
after	O
CD80	B-protein
compared	O
to	O
CD86	B-protein
ligation	O
.	O

Our	O
results	O
suggest	O
that	O
the	O
eventual	O
appearance	O
of	O
CD80	B-protein
on	O
recently	O
activated	O
CD86+	B-cell_type
APC	I-cell_type
is	O
important	O
for	O
the	O
superinduction	O
of	O
IL-2	B-protein
production	O
and	O
to	O
support	O
vigorous	O
T	O
cell	O
proliferation	O
.	O

-DOCSTART-	O

Duplication	O
of	O
the	O
DR3	B-DNA
gene	I-DNA
on	O
human	B-DNA
chromosome	I-DNA
1p36	I-DNA
and	O
its	O
deletion	O
in	O
human	B-cell_type
neuroblastoma	I-cell_type
.	O

The	O
human	B-DNA
DR3	I-DNA
gene	I-DNA
,	O
whose	O
product	O
is	O
also	O
known	O
as	O
Wsl-1/APO-3/TRAMP/LARD	B-protein
,	O
encodes	O
a	O
tumor	B-protein
necrosis	I-protein
factor-related	I-protein
receptor	I-protein
that	O
is	O
expressed	O
primarily	O
on	O
the	O
surface	O
of	O
thymocytes	B-cell_type
and	O
lymphocytes	B-cell_type
.	O

DR3	B-protein
is	O
capable	O
of	O
inducing	O
both	O
NF-kappa	B-protein
B	I-protein
activation	O
and	O
apoptosis	O
when	O
overexpressed	O
in	O
mammalian	B-cell_type
cells	I-cell_type
,	O
although	O
its	O
ligand	O
has	O
not	O
yet	O
been	O
identified	O
.	O

We	O
report	O
here	O
that	O
the	O
DR3	B-DNA
gene	I-DNA
locus	I-DNA
is	O
tandemly	O
duplicated	O
on	O
human	B-DNA
chromosome	I-DNA
band	I-DNA
1p36.2-p36.3	I-DNA
and	O
that	O
these	O
genes	O
are	O
hemizygously	O
deleted	O
and/or	O
translocated	O
to	O
another	O
chromosome	O
in	O
neuroblastoma	B-cell_line
(	I-cell_line
NB	I-cell_line
)	I-cell_line
cell	I-cell_line
lines	I-cell_line
with	O
amplified	O
MYCN	O
.	O

Duplication	O
of	O
at	O
least	O
a	O
portion	O
of	O
the	O
DR3	B-DNA
gene	I-DNA
,	O
including	O
the	O
extracellular	B-protein
and	I-protein
transmembrane	I-protein
regions	I-protein
but	O
not	O
the	O
cytoplasmic	B-protein
domain	I-protein
,	O
was	O
demonstrated	O
by	O
both	O
fluorescence	O
in	O
situ	O
hybridization	O
and	O
genomic	O
Southern	O
blotting	O
.	O

In	O
most	O
NB	B-cell_line
cell	I-cell_line
lines	I-cell_line
,	O
both	O
the	O
DR3	B-DNA
and	I-DNA
the	I-DNA
DR3L	I-DNA
sequences	I-DNA
are	O
simultaneously	O
deleted	O
and/or	O
translocated	O
to	O
another	O
chromosome	B-DNA
.	O

Finally	O
,	O
DR3/	B-protein
Wsl-1	I-protein
protein	I-protein
expression	O
is	O
quite	O
variable	O
among	O
these	O
NB	B-cell_line
cell	I-cell_line
lines	I-cell_line
,	O
with	O
very	O
low	O
or	O
undetectable	O
levels	O
in	O
7	O
of	O
17	O
NB	B-cell_line
cell	I-cell_line
lines	I-cell_line

-DOCSTART-	O

Differential	O
protection	O
of	O
normal	B-cell_type
and	I-cell_type
malignant	I-cell_type
human	I-cell_type
myeloid	I-cell_type
progenitors	I-cell_type
(	O
CFU-GM	B-cell_type
)	O
from	O
Ara-C	O
toxicity	O
using	O
cycloheximide	O
.	O

Cycloheximide	O
,	O
a	O
reversible	O
protein	O
synthesis	O
inhibitor	O
,	O
is	O
thought	O
to	O
block	O
DNA	O
replication	O
in	O
normal	B-cell_type
cells	I-cell_type
by	O
preventing	O
synthesis	O
of	O
a	O
labile	B-protein
protein	I-protein
.	O

In	O
animal	O
systems	O
,	O
cycloheximide	O
protects	O
normal	B-cell_type
cells	I-cell_type
from	O
cytotoxic	O
S-phase	O
specific	O
agents	O
,	O
such	O
as	O
cytosine	O
arabinoside	O
(	O
Ara-C	O
)	O
.	O

Malignant	B-cell_type
cells	I-cell_type
appear	O
not	O
to	O
be	O
susceptible	O
to	O
cycloheximide-induced	O
cycle	O
arrest	O
and	O
,	O
subsequently	O
,	O
may	O
not	O
be	O
protected	O
from	O
Ara-C	O
cytotoxicity	O
.	O

The	O
effect	O
of	O
cycloheximide	O
on	O
granulocyte/macrophage	B-cell_type
progenitors	I-cell_type
(	O
CFU-GM	B-cell_type
)	O
after	O
in	O
vitro	O
Ara-C	O
exposure	O
was	O
examined	O
using	O
normal	O
human	O
bone	O
marrow	O
,	O
malignant	B-cell_type
progenitors	I-cell_type
from	O
patients	O
with	O
chronic	O
myelogenous	O
leukemia	O
(	O
CML	O
)	O
,	O
and	O
clonogenic	B-cell_line
cells	I-cell_line
from	O
the	O
human	B-cell_line
acute	I-cell_line
nonlymphocytic	I-cell_line
leukemia	I-cell_line
cell	I-cell_line
lines	I-cell_line
HL-60	I-cell_line
and	I-cell_line
KG-1	I-cell_line
.	O

Mononuclear	B-cell_line
or	I-cell_line
clonogenic	I-cell_line
cells	I-cell_line
were	O
incubated	O
for	O
one	O
hour	O
with	O
cycloheximide	O
,	O
followed	O
by	O
the	O
addition	O
,	O
for	O
three	O
or	O
17	O
hours	O
,	O
of	O
Ara-C	O
before	O
being	O
plated	O
in	O
a	O
methylcellulose	O
culture	O
system	O
.	O

CFU-GM	O
survival	O
was	O
significantly	O
increase	O
if	O
normal	O
cells	O
were	O
treated	O
with	O
cycloheximide	O
before	O
Ara-C	O
exposure	O
.	O

Similar	O
cycloheximide	O
pretreatment	O
of	O
CML	B-cell_type
progenitors	I-cell_type
and	O
clonogenic	B-cell_line
HL-60	I-cell_line
and	I-cell_line
KG-1	I-cell_line
cells	I-cell_line
failed	O
to	O
protect	O
CFU-GM	B-cell_type
from	O
Ara-C-induced	O
cytotoxicity	O
.	O

-DOCSTART-	O

Primary	O
cortisol	O
resistance	O
accompanied	O
by	O
a	O
reduction	O
in	O
glucocorticoid	B-protein
receptors	I-protein
in	O
two	O
members	O
of	O
the	O
same	O
family	O
.	O

This	O
report	O
describes	O
studies	O
of	O
a	O
man	O
suspected	O
of	O
having	O
primary	O
cortisol	O
resistance	O
.	O

This	O
conclusion	O
is	O
based	O
on	O
his	O
high	O
plasma	O
cortisol	O
levels	O
and	O
high	O
24-h	O
urinary	O
17-hydroxycorticosteroid	O
and	O
cortisol	O
excretion	O
,	O
plus	O
the	O
fact	O
that	O
he	O
had	O
no	O
manifestations	O
of	O
Cushing	O
's	O
syndrome	O
.	O

Among	O
family	O
members	O
tested	O
,	O
his	O
mother	O
also	O
had	O
hypercortisolemia	O
.	O

Both	O
mother	O
and	O
son	O
had	O
high	O
levels	O
of	O
unbound	O
plasma	O
cortisol	O
,	O
but	O
their	O
plasma	O
ACTH	O
concentrations	O
were	O
within	O
the	O
normal	O
range	O
.	O

Both	O
were	O
partially	O
resistant	O
to	O
dexamethasone	O
adrenal	O
suppression	O
,	O
and	O
both	O
had	O
mild	O
hypertension	O
without	O
hypokalemia	O
.	O

To	O
study	O
this	O
apparent	O
end-organ	O
resistance	O
to	O
cortisol	O
,	O
we	O
examined	O
the	O
glucocorticoid	B-protein
receptors	I-protein
in	O
peripheral	B-cell_type
mononuclear	I-cell_type
cells	I-cell_type
.	O

Using	O
whole	O
cell	O
assays	O
,	O
glucocorticoid	B-protein
receptors	I-protein
in	O
both	O
patients	O
were	O
found	O
to	O
have	O
reduced	O
total	O
binding	O
capacity	O
.	O

We	O
conclude	O
that	O
these	O
two	O
patients	O
,	O
members	O
of	O
the	O
same	O
family	O
,	O
have	O
primary	O
cortisol	O
resistance	O
accompanied	O
by	O
a	O
reduced	O
number	O
of	O
glucocorticoid	B-protein
receptors	I-protein
.	O

-DOCSTART-	O

[	O
Glucocorticoid	B-protein
receptors	I-protein
and	O
response	O
to	O
polychemotherapy	O
in	O
acute	O
lymphatic	O
leukemia	O
]	O

Glucocorticoid	B-protein
receptor	I-protein
(	O
GR	B-protein
)	O
levels	O
were	O
quantified	O
in	O
leukemic	B-cell_type
blasts	I-cell_type
from	O
peripheral	O
blood	O
of	O
86	O
patients	O
with	O
acute	O
lymphoblastic	O
leukemia	O
.	O

The	O
subsequent	O
achievement	O
of	O
complete	O
remission	O
after	O
combination	O
chemotherapy	O
was	O
correlated	O
with	O
high	O
receptor	O
levels	O
.	O

Forty-seven	O
of	O
50	O
patients	O
with	O
leukemic	B-cell_type
cells	I-cell_type
containing	O
more	O
than	O
6	O
,	O
000	O
receptor	O
sites	O
and	O
22	O
of	O
36	O
patients	O
with	O
cells	O
containing	O
less	O
than	O
6	O
,	O
000	O
receptor	O
sites	O
achieved	O
remission	O
.	O

The	O
study	O
of	O
glucocorticoid	B-protein
receptors	I-protein
in	O
leukemic	B-cell_type
cells	I-cell_type
may	O
predict	O
response	O
to	O
combination	O
chemotherapy	O
in	O
patients	O
with	O
acute	O
lymphoblastic	O
leukemia	O
.	O

-DOCSTART-	O

Specific	O
uptake	O
of	O
1	O
,	O
25-dihydroxycholecalciferol	O
by	O
human	B-cell_type
chronic	I-cell_type
myeloid	I-cell_type
leukemia	I-cell_type
cells	I-cell_type
.	O

We	O
have	O
examined	O
mononuclear	B-cell_line
cell	I-cell_line
preparations	I-cell_line
from	O
patients	O
with	O
chronic	O
myeloid	O
leukemia	O
[	O
CML	O
]	O
for	O
binding	O
of	O
and	O
response	O
to	O
1	O
,	O
25-dihydroxycholecalciferol	O
[	O
1	O
,	O
25-	O
(	O
OH	O
)	O
2D3	O
]	O
.	O

Whole	O
cells	O
specifically	O
took	O
up	O
[	O
3H	O
]	O
-1	O
,	O
25-	O
(	O
OH	O
)	O
2D3	O
with	O
high	O
affinity	O
(	O
Kd	O
3.6	O
X	O
10	O
(	O
-11	O
)	O
M	O
)	O
and	O
low	O
capacity	O
.	O

Subcellular	O
fractionation	O
of	O
labeled	B-cell_line
cells	I-cell_line
showed	O
that	O
binding	O
was	O
restricted	O
to	O
cytosols	O
and	O
nuclei	O
.	O

Sucrose	O
gradient	O
centrifugation	O
of	O
cells	O
preincubated	O
with	O
[	O
3H	O
]	O
-1	O
,	O
25-	O
(	O
OH	O
)	O
2D3	O
revealed	O
a	O
single	O
3.6S	O
peak	O
which	O
was	O
totally	O
displaced	O
with	O
100-fold	O
excess	O
nonradioactive	O
hormone	O
.	O

However	O
,	O
we	O
were	O
unable	O
to	O
demonstrate	O
specific	O
binding	O
of	O
1	O
,	O
25-	O
(	O
OH	O
)	O
2D3	O
by	O
postlabeling	O
standard	O
cytosol	O
preparations	O
.	O

In	O
addition	O
,	O
cytosols	O
prepared	O
from	O
a	O
mixture	O
of	O
CML	B-cell_line
cells	I-cell_line
and	O
1	B-cell_line
,	I-cell_line
25-	I-cell_line
(	I-cell_line
OH	I-cell_line
)	I-cell_line
2D3	I-cell_line
receptor-positive	I-cell_line
T47D	I-cell_line
(	O
human	O
breast	O
cancer	O
)	O
cells	O
had	O
less	O
than	O
10	O
%	O
of	O
the	O
binding	O
measured	O
in	O
T47D	B-cell_line
cytosol	O
alone	O
.	O

However	O
,	O
the	O
levels	O
of	O
binding	O
in	O
T47D	B-cell_line
cytosols	O
were	O
not	O
reduced	O
if	O
the	O
receptors	O
were	O
occupied	O
with	O
[	O
3H	O
]	O
-1	O
,	O
25-	O
(	O
OH	O
)	O
2D3	O
prior	O
to	O
the	O
addition	O
of	O
the	O
CML	B-cell_line
cytosols	O
.	O

Thus	O
,	O
CML	B-cell_line
cells	I-cell_line
appear	O
to	O
contain	O
both	O
the	O
receptor	O
for	O
1	O
,	O
25-	O
(	O
OH	O
)	O
2D3	O
and	O
an	O
unknown	O
substance	O
which	O
prevents	O
its	O
detection	O
following	O
the	O
preparation	O
of	O
cytosol	O
.	O

Cells	O
from	O
patients	O
with	O
CML	O
in	O
the	O
chronic	O
phase	O
specifically	O
bound	O
more	O
1	O
,	O
25-	O
(	O
OH	O
)	O
2D3	O
[	O
18.0	O
+/-	O
3.2	O
(	O
S.E.	O
)	O
fmol/10	O
(	O
7	O
)	O
cells	O
]	O
than	O
did	O
those	O
in	O
acute	O
myeloid	O
transformation	O
[	O
7.2	O
+/-	O
1.5	O
]	O
or	O
than	O
did	O
cells	O
from	O
patients	O
with	O
acute	O
myeloid	O
leukemia	O
[	O
2.6	O
+/-	O
0.8	O
]	O
.	O

Only	O
cells	O
from	O
the	O
first	O
group	O
of	O
patients	O
responded	O
to	O
the	O
addition	O
of	O
1	O
,	O
25-	O
(	O
OH	O
)	O
2D3	O
by	O
differentiating	O
along	O
the	O
monocyte-macrophage	O
pathway	O
.	O

We	O
conclude	O
that	O
the	O
differentiation-induction	O
effect	O
of	O
1	O
,	O
25-	O
(	O
OH	O
)	O
2D3	O
is	O
likely	O
to	O
depend	O
on	O
adequate	O
levels	O
of	O
receptor	O
and	O
that	O
intact	B-cell_type
cells	I-cell_type
rather	O
than	O
cytosol	O
preparations	O
should	O
be	O
studied	O
before	O
cells	O
of	O
a	O
particular	O
tissue	O
are	O
designated	O
as	O
receptor	O
negative	O
.	O

-DOCSTART-	O

Effect	O
of	O
cell	O
cycle	O
position	O
on	O
dexamethasone	O
binding	O
by	O
mouse	B-cell_line
and	I-cell_line
human	I-cell_line
lymphoid	I-cell_line
cell	I-cell_line
lines	I-cell_line
:	O
correlation	O
between	O
an	O
increase	O
in	O
dexamethasone	O
binding	O
during	O
S	O
phase	O
and	O
dexamethasone	O
sensitivity	O
.	O

We	O
determined	O
the	O
effect	O
of	O
cell	O
cycle	O
position	O
on	O
the	O
amount	O
of	O
dexamethasone	O
that	O
was	O
specifically	O
bound	O
by	O
mouse	B-cell_line
and	I-cell_line
human	I-cell_line
lymphoid	I-cell_line
cell	I-cell_line
lines	I-cell_line
.	O

Cell	B-cell_line
lines	I-cell_line
that	O
were	O
either	O
sensitive	O
or	O
resistant	O
to	O
growth	O
inhibition	O
by	O
dexamethasone	O
were	O
compared	O
.	O

Exponentially	B-cell_type
growing	I-cell_type
cells	I-cell_type
were	O
separated	O
by	O
centrifugal	O
elutriation	O
into	O
fractions	O
that	O
corresponded	O
to	O
different	O
positions	O
in	O
the	O
cell	O
cycle	O
.	O

The	O
cell	O
cycle	O
phase	O
distribution	O
of	O
each	O
fraction	O
was	O
estimated	O
by	O
flow	O
cytometry	O
and	O
autoradiography	O
.	O

The	O
amount	O
of	O
dexamethasone	O
bound	O
per	O
cell	O
in	O
each	O
fraction	O
was	O
measured	O
by	O
a	O
whole	O
cell	O
binding	O
assay	O
.	O

In	O
three	O
dexamethasone-sensitive	B-cell_line
cell	I-cell_line
lines	I-cell_line
(	O
two	O
mouse	O
and	O
one	O
human	O
)	O
,	O
we	O
found	O
that	O
the	O
amount	O
of	O
dexamethasone	O
bound	O
per	O
cell	O
increased	O
2-4-fold	O
between	O
G1	O
phase	O
and	O
S	O
phase	O
,	O
and	O
then	O
decreased	O
during	O
G2/M	O
phase	O
.	O

Results	O
were	O
the	O
same	O
when	O
the	O
amount	O
of	O
dexamethasone	O
bound	O
per	O
milligram	O
of	O
cell	B-protein
protein	I-protein
was	O
measured	O
.	O

Binding	O
affinity	O
was	O
the	O
same	O
during	O
G1	O
phase	O
and	O
S	O
phase	O
,	O
but	O
the	O
proportion	O
of	O
bound	O
dexamethasone	O
that	O
translocated	O
to	O
the	O
nucleus	O
was	O
greater	O
during	O
S	O
phase	O
.	O

In	O
contrast	O
,	O
we	O
found	O
that	O
the	O
amount	O
of	O
dexamethasone	O
bound	O
per	O
cell	O
by	O
three	O
dexamethasone-resistant	B-cell_line
cell	I-cell_line
lines	I-cell_line
(	O
two	O
mouse	O
and	O
one	O
human	O
)	O
did	O
not	O
increase	O
during	O
S	O
phase	O
.	O

Our	O
results	O
indicate	O
that	O
cell	O
cycle	O
changes	O
in	O
dexamethasone	O
binding	O
are	O
not	O
simply	O
related	O
to	O
changes	O
in	O
cell	B-protein
protein	I-protein
or	O
cell	O
volume	O
during	O
the	O
cell	O
cycle	O
.	O

An	O
increase	O
in	O
dexamethasone	O
binding	O
during	O
S	O
phase	O
may	O
be	O
required	O
for	O
dexamethasone	O
to	O
inhibit	O
cell	O
growth	O
,	O
and	O
a	O
failure	O
of	O
dexamethasone	O
binding	O
to	O
increase	O
during	O
S	O
phase	O
might	O
represent	O
a	O
new	O
mechanism	O
of	O
dexamethasone	O
resistance	O
in	O
lymphoid	B-cell_type
cells	I-cell_type
.	O

-DOCSTART-	O

Presence	O
and	O
steroid	O
inducibility	O
of	O
glutamine	B-protein
synthetase	I-protein
in	O
human	B-cell_type
leukemic	I-cell_type
cells	I-cell_type
.	O

Glutamine	B-protein
synthetase	I-protein
(	O
EC	O
6.3.1.2	O
;	O
GS	O
)	O
is	O
present	O
in	O
lymphoblasts	B-cell_type
from	O
patients	O
with	O
acute	O
lymphoblastic	O
leukemia	O
(	O
ALL	O
)	O
as	O
well	O
as	O
in	O
normal	B-cell_type
peripheral	I-cell_type
blood	I-cell_type
lymphocytes	I-cell_type
.	O

In	O
16	O
out	O
of	O
20	O
ALL	O
patients	O
studied	O
exposure	O
of	O
the	O
cells	O
to	O
physiological	O
concentrations	O
of	O
dexamethasone	O
in	O
vitro	O
increased	O
enzyme	B-protein
activity	O
above	O
the	O
control	O
levels	O
.	O

The	O
increase	O
was	O
specific	O
for	O
glucocorticoid	B-protein
receptor	I-protein
ligands	O
.	O

A	O
direct	O
correlation	O
was	O
found	O
between	O
the	O
magnitude	O
of	O
glucocorticoid-mediated	O
increase	O
of	O
GS	O
activity	O
and	O
the	O
cellular	O
levels	O
of	O
specific	O
glucocorticoid	B-protein
receptors	I-protein
assayed	O
in	O
the	O
same	O
cell	B-cell_type
specimen	I-cell_type
.	O

Moreover	O
,	O
the	O
basal	O
levels	O
of	O
the	O
enzyme	B-protein
measured	O
in	O
cells	O
prior	O
to	O
exposure	O
to	O
dexamethasone	O
correlated	O
negatively	O
with	O
receptor	O
density	O
.	O

It	O
is	O
suggested	O
that	O
the	O
presence	O
of	O
steroid-inducible	B-protein
GS	I-protein
in	O
ALL	B-cell_type
cells	I-cell_type
may	O
prove	O
to	O
be	O
a	O
marker	O
for	O
functional	B-protein
receptor	I-protein
sites	I-protein
.	O

-DOCSTART-	O

Binding	O
of	O
progestins	O
to	O
the	O
glucocorticoid	B-protein
receptor	I-protein
.	O

Correlation	O
to	O
their	O
glucocorticoid-like	O
effects	O
on	O
in	O
vitro	O
functions	O
of	O
human	B-cell_type
mononuclear	I-cell_type
leukocytes	I-cell_type
.	O

A	O
number	O
of	O
physiological	O
and	O
synthetic	O
progestins	O
were	O
tested	O
for	O
their	O
ability	O
to	O
compete	O
with	O
[	O
3H	O
]	O
dexamethasone	O
for	O
the	O
binding	O
to	O
the	O
glucocorticoid	B-protein
receptor	I-protein
of	O
human	B-cell_type
mononuclear	I-cell_type
leukocytes	I-cell_type
and	O
their	O
ability	O
to	O
elicit	O
glucocorticoid-like	O
effects	O
on	O
the	O
same	O
cells	O
.	O

As	O
compared	O
to	O
the	O
reference	O
compound	O
dexamethasone	O
(	O
relative	O
receptor	O
binding	O
affinity	O
defined	O
as	O
100	O
%	O
)	O
,	O
two	O
potent	O
synthetic	O
progestins	O
with	O
a	O
pregnane-type	O
structure	O
,	O
megestrol	O
acetate	O
and	O
medroxyprogesterone	O
acetate	O
,	O
were	O
found	O
to	O
display	O
a	O
considerable	O
binding	O
affinity	O
towards	O
the	O
receptor	O
(	O
46	O
and	O
42	O
%	O
,	O
respectively	O
)	O
.	O

The	O
relative	O
binding	O
affinity	O
of	O
the	O
naturally	O
occurring	O
ligand	O
,	O
cortisol	O
,	O
to	O
the	O
receptor	O
was	O
clearly	O
lower	O
(	O
25	O
%	O
)	O
.	O

The	O
effective	O
binding	O
of	O
medroxyprogesterone	O
acetate	O
to	O
the	O
glucocorticoid	B-protein
receptor	I-protein
was	O
confirmed	O
by	O
direct	O
binding	O
studies	O
utilizing	O
a	O
tritiated	O
derivative	O
of	O
this	O
steroid	O
.	O

No	O
evidence	O
for	O
the	O
existence	O
of	O
a	O
specific	O
progesterone	B-protein
receptor	I-protein
in	O
human	B-cell_type
mononuclear	I-cell_type
leukocytes	I-cell_type
was	O
obtained	O
as	O
judged	O
by	O
the	O
results	O
of	O
competition	O
experiments	O
where	O
a	O
progesterone	O
receptor-specific	O
ligand	O
[	O
3H	O
]	O
Org	O
2058	O
was	O
used	O
.	O

Medroxyprogesterone	O
acetate	O
and	O
megestrol	O
acetate	O
also	O
induced	O
glucocorticoid-like	O
effects	O
on	O
the	O
lymphocyte	B-cell_type
functions	O
.	O

These	O
included	O
inhibition	O
of	O
the	O
proliferative	O
responses	O
to	O
the	O
T-cell	B-protein
mitogens	I-protein
concanavalin	B-protein
A	I-protein
and	O
phytohaemagglutinin	B-protein
and	O
an	O
enhanced	O
accumulation	O
of	O
immunoglobulin	B-cell_type
secreting	I-cell_type
cells	I-cell_type
in	O
pokeweed	B-cell_line
mitogen-stimulated	I-cell_line
cultures	I-cell_line
.	O

The	O
progestin	O
effect	O
appears	O
to	O
be	O
mediated	O
through	O
a	O
radiosensitive	O
(	O
suppressor	O
)	O
subpopulation	O
of	O
T	B-cell_type
lymphocytes	I-cell_type
.	O

In	O
contrast	O
,	O
the	O
synthetic	O
progestins	O
related	O
structurally	O
to	O
19-nortestosterone	O
,	O
norethisterone	O
and	O
d-norgestrel	O
,	O
were	O
virtually	O
devoid	O
of	O
binding	O
affinity	O
towards	O
the	O
glucocorticoid	B-protein
receptor	I-protein
nor	O
did	O
they	O
measurably	O
influence	O
the	O
in	O
vitro	O
lymphocyte	B-cell_type
functions	O
.	O

These	O
studies	O
demonstrate	O
that	O
certain	O
progestins	O
in	O
common	O
clinical	O
use	O
probably	O
possess	O
inherent	O
glucocorticoid	O
activity	O
and	O
suggest	O
that	O
side	O
effects	O
attributable	O
to	O
this	O
character	O
(	O
e.g.	O
suppression	O
of	O
the	O
pituitary-adrenal	O
axis	O
)	O
might	O
be	O
expected	O
when	O
these	O
compounds	O
are	O
used	O
in	O
pharmacological	O
doses	O
.	O

-DOCSTART-	O

Metabolic	O
and	O
ultrastructural	O
aspects	O
of	O
the	O
in	O
vitro	O
lysis	O
of	O
chronic	B-cell_type
lymphocytic	I-cell_type
leukemia	I-cell_type
cells	I-cell_type
by	O
glucocorticoids	O
.	O

Human	B-cell_type
chronic	I-cell_type
lymphocytic	I-cell_type
leukemia	I-cell_type
(	I-cell_type
CLL	I-cell_type
)	I-cell_type
cells	I-cell_type
like	O
prothymocytes	B-cell_type
and	O
immunoactivated	B-cell_type
T-lymphocytes	I-cell_type
are	O
readily	O
lysed	O
in	O
vitro	O
by	O
pharmacological	O
concentrations	O
of	O
glucocorticoids	O
such	O
as	O
cortisol	O
,	O
whereas	O
peripheral	B-cell_type
blood	I-cell_type
lymphocytes	I-cell_type
and	O
thymocytes	B-cell_type
are	O
unaffected	O
by	O
the	O
hormone	B-protein
.	O

In	O
this	O
study	O
,	O
metabolic	O
and	O
ultrastructural	O
aspects	O
of	O
the	O
cortisol-induced	O
killing	O
process	O
of	O
CLL	B-cell_type
cells	I-cell_type
are	O
recorded	O
.	O

In	O
vitro	O
lysis	O
was	O
found	O
to	O
be	O
temperature	O
dependent	O
and	O
was	O
detected	O
only	O
after	O
6	O
to	O
8	O
hr	O
incubation	O
with	O
cortisol	O
by	O
means	O
of	O
the	O
trypan	O
blue	O
exclusion	O
test	O
.	O

However	O
,	O
30	O
min	O
of	O
incubation	O
with	O
cortisol	O
at	O
either	O
37	O
degrees	O
or	O
4	O
degrees	O
followed	O
by	O
the	O
removal	O
of	O
the	O
hormone	B-protein
was	O
still	O
sufficient	O
to	O
induce	O
the	O
lytic	O
process	O
.	O

Ultrastructural	O
studies	O
demonstrated	O
sequential	O
changes	O
in	O
the	O
cytoplasm	O
,	O
including	O
swelling	O
of	O
mitochondria	O
and	O
cytoplasmic	O
decompartmentalization	O
,	O
followed	O
by	O
loss	O
of	O
surface	O
microvilli	O
with	O
the	O
appearance	O
of	O
``	O
holes	O
''	O
in	O
the	O
cell	O
membrane	O
,	O
and	O
subsequent	O
condensation	O
of	O
nuclear	B-DNA
chromatin	I-DNA
.	O

The	O
large	O
holes	O
in	O
the	O
membrane	O
appearing	O
after	O
6	O
hr	O
of	O
incubation	O
with	O
the	O
hormone	B-protein
may	O
be	O
the	O
cause	O
for	O
the	O
penetration	O
of	O
the	O
viable	O
stain	O
into	O
the	O
dead	B-cell_type
cells	I-cell_type
,	O
as	O
seen	O
by	O
light	O
microscopy	O
.	O

Addition	O
of	O
metabolic	O
inhibitors	O
including	O
actinomycin	O
D	O
,	O
puromycin	O
,	O
and	O
cycloheximide	O
following	O
administration	O
of	O
cortisol	O
resulted	O
in	O
inhibition	O
of	O
the	O
cell	O
lysis	O
.	O

An	O
excess	O
of	O
an	O
antagonist	O
such	O
as	O
cortexolone	O
was	O
found	O
to	O
inhibit	O
the	O
cortisol-induced	O
cytolysis	O
of	O
the	O
CLL	B-cell_line
cells	I-cell_line
.	O

It	O
is	O
suggested	O
that	O
the	O
glucocorticoid-induced	O
lysis	O
of	O
human	B-cell_line
CLL	I-cell_line
cells	I-cell_line
is	O
similar	O
to	O
the	O
phenomenon	O
observed	O
in	O
rat	B-cell_type
or	I-cell_type
murine	I-cell_type
lymphocytes	I-cell_type
and	O
is	O
mediated	O
by	O
interaction	O
of	O
the	O
steroid	O
molecule	O
with	O
the	O
cytoplasmic	B-protein
receptor	I-protein
.	O

The	O
resulting	O
complex	O
appears	O
to	O
activate	O
specific	O
gene	B-DNA
(	O
s	O
)	O
the	O
products	O
of	O
which	O
eventually	O
cause	O
cytolysis	O
.	O

-DOCSTART-	O

Glucocorticoid	B-protein
receptors	I-protein
in	O
lymphoid	O
tumors	O
.	O

There	O
is	O
a	O
range	O
of	O
levels	O
of	O
glucocorticoid	B-protein
receptor	I-protein
numbers	O
seen	O
in	O
the	O
various	O
subclasses	O
of	O
acute	O
lymphatic	O
leukemia	O
(	O
ALL	O
)	O
.	O

This	O
variability	O
can	O
not	O
be	O
explained	O
by	O
the	O
known	O
correlation	O
between	O
active	O
cell	O
proliferation	O
and	O
an	O
increase	O
in	O
the	O
number	O
of	O
receptors	O
,	O
since	O
the	O
tumors	O
with	O
the	O
highest	O
growth	O
fraction	O
(	O
i.e.	O
,	O
Burkitt	O
's	O
lymphoma	O
and	O
T-cell	O
leukemia	O
)	O
tend	O
to	O
have	O
lower	O
average	O
receptor	O
numbers	O
than	O
do	O
tumors	O
with	O
lower	O
growth	O
fractions	O
such	O
as	O
common	O
ALL	O
.	O

All	O
clinical	O
specimens	O
from	O
patients	O
with	O
lymphatic	O
leukemia	O
have	O
some	O
measurable	O
level	O
of	O
glucocorticoid	B-protein
receptors	I-protein
;	O
therefore	O
,	O
the	O
resistance	O
seen	O
in	O
vivo	O
can	O
not	O
be	O
explained	O
by	O
the	O
lack	O
of	O
receptors	O
.	O

However	O
,	O
there	O
has	O
been	O
a	O
positive	O
correlation	O
,	O
in	O
our	O
hands	O
,	O
with	O
receptor	O
level	O
and	O
prognosis	O
.	O

On	O
the	O
basis	O
of	O
in	O
vitro	O
models	O
,	O
it	O
is	O
proposed	O
that	O
perhaps	O
the	O
high	B-cell_line
receptor	I-cell_line
cell	I-cell_line
lines	I-cell_line
(	O
i.e.	O
,	O
common	O
ALL	O
of	O
childhood	O
)	O
have	O
relative	O
stability	O
of	O
their	O
genetic	O
material	O
making	O
glucocorticoid-resistant	O
mutations	O
less	O
likely	O
to	O
occur	O
in	O
patients	O
with	O
these	O
cells	O
than	O
in	O
low-receptor	B-cell_line
cell	I-cell_line
lines	I-cell_line
(	O
i.e.	O
,	O
T-cell	O
leukemia	O
)	O
.	O

This	O
greater	O
genetic	O
variability	O
in	O
the	O
low-receptor	B-cell_line
lines	I-cell_line
could	O
account	O
for	O
the	O
earlier	O
emergence	O
of	O
clinical	O
glucocorticoid	O
resistance	O
in	O
these	O
patients	O
.	O

-DOCSTART-	O

[	O
Tumor	O
histology	O
and	O
steroid	B-protein
receptors	I-protein
in	O
breast	O
carcinoma	O
]	O

In	O
Specimens	O
of	O
115	O
patients	O
with	O
breast	O
cancer	O
4	O
tumorparameters	O
(	O
tumorsize	O
,	O
tumorboder	O
,	O
nucleargrade	O
,	O
lymphocytic	O
stromal	O
reaction	O
)	O
3	O
features	O
of	O
regional	O
lymphnodes	O
(	O
sinushistiocytosis	O
,	O
T-cellreaction	O
,	O
lymphnode	O
metastases	O
)	O
and	O
estrogen	B-protein
and	I-protein
progesteron	I-protein
receptors	I-protein
were	O
determined	O
.	O

A	O
strong	O
sinushistiocytosis	O
and	O
T-cellreaction	O
could	O
be	O
verified	O
mainly	O
in	O
metastases	O
in	O
free	O
lymphnodes	O
.	O

The	O
steroid	B-protein
receptor	I-protein
content	O
does	O
not	O
correlate	O
with	O
histological	O
parameters	O

-DOCSTART-	O

Aldosterone-receptor	B-protein
deficiency	O
in	O
pseudohypoaldosteronism	O
.	O

Pseudohypoaldosteronism	O
,	O
a	O
syndrome	O
characterized	O
by	O
salt	O
wasting	O
and	O
failure	O
to	O
thrive	O
,	O
usually	O
presents	O
in	O
infancy	O
as	O
high	O
urinary	O
levels	O
of	O
sodium	O
despite	O
hyponatremia	O
,	O
hyperkalemia	O
,	O
hyperreninemia	O
,	O
and	O
elevated	O
aldosterone	O
levels	O
.	O

We	O
have	O
investigated	O
this	O
syndrome	O
for	O
the	O
possibility	O
of	O
abnormal	B-protein
Type	I-protein
I	I-protein
or	O
``	B-protein
mineralocorticoid-like	I-protein
''	I-protein
receptors	I-protein
,	O
which	O
have	O
intrinsic	O
steroid	O
specificity	O
indistinguishable	O
from	O
that	O
of	O
renal	B-protein
mineralocorticoid	I-protein
receptors	I-protein
and	O
are	O
found	O
in	O
many	O
tissues	O
and	O
cells	B-cell_type
,	O
including	O
mononuclear	B-cell_type
leukocytes	I-cell_type
.	O

We	O
have	O
studied	O
three	O
patients	O
with	O
pseudohypoaldosteronism	O
:	O
the	O
28-year-old	O
index	O
case	O
in	O
Melbourne	O
(	O
Patient	O
1	O
)	O
and	O
two	O
siblings	O
in	O
Munich	O
,	O
eight	O
and	O
two	O
years	O
of	O
age	O
(	O
Patients	O
2	O
and	O
3	O
)	O
;	O
clinically	O
,	O
Patient	O
3	O
had	O
a	O
less	O
severe	O
case	O
than	O
his	O
sister	O
.	O

Percoll-separated	B-cell_type
control	I-cell_type
monocytes	I-cell_type
bound	O
[	O
3H	O
]	O
aldosterone	O
with	O
high	O
affinity	O
(	O
Kd	O
approximately	O
3	O
nM	O
)	O
and	O
limited	O
capacity	O
(	O
150	O
to	O
600	O
sites	O
per	O
cell	O
)	O
.	O

On	O
repeated	O
examination	O
,	O
no	O
[	O
3H	O
]	O
aldosterone	O
binding	O
was	O
found	O
in	O
monocytes	B-cell_type
from	O
Patients	O
1	O
and	O
2	O
;	O
in	O
Patient	O
3	O
,	O
the	O
levels	O
were	O
62	O
sites	O
per	O
cell	O
,	O
more	O
than	O
2	O
S.D.	O
below	O
those	O
of	O
the	O
control	O
.	O

Levels	O
in	O
the	O
parents	O
of	O
the	O
Munich	O
patients	O
(	O
first	O
cousins	O
)	O
were	O
normal	O
.	O

It	O
appears	O
that	O
pseudohypoaldosteronism	O
is	O
caused	O
by	O
a	O
Type	O
I	O
receptor	O
defect	O
,	O
that	O
the	O
defect	O
may	O
be	O
complete	O
or	O
partial	O
,	O
that	O
transmission	O
may	O
be	O
autosomal	O
recessive	O
,	O
and	O
that	O
the	O
study	O
of	O
patients	O
with	O
pseudohypoaldosteronism	O
may	O
indicate	O
physiologic	O
roles	O
for	O
Type	B-protein
I	I-protein
receptors	I-protein
in	O
nonepithelial	O
tissues	O
.	O

-DOCSTART-	O

Glucocorticoid	O
inhibition	O
of	O
urokinase-like	B-protein
plasminogen	I-protein
activators	I-protein
in	O
cultured	B-cell_line
human	I-cell_line
lymphoblasts	I-cell_line
.	O

Two	O
human	B-cell_line
lymphoblast	I-cell_line
cell	I-cell_line
lines	I-cell_line
,	O
LICR-LON-HMy2	B-cell_line
(	O
HMy2	B-cell_line
cells	I-cell_line
)	O
and	O
GM4672A	B-cell_line
cells	I-cell_line
,	O
are	O
moderately	O
growth	O
inhibited	O
by	O
dexamethasone	O
(	O
1	O
,	O
4-pregnadien-9-fluoro-16	O
alpha-methyl-11	O
beta	O
,	O
17	O
alpha	O
,	O
21-triol-3	O
,	O
20-dione	O
)	O
(	O
Dex	O
)	O
.	O

Both	O
cell	O
types	O
secrete	O
a	O
urokinase	B-protein
(	I-protein
UK	I-protein
)	I-protein
-like	I-protein
plasminogen	I-protein
activator	I-protein
(	O
PA	B-protein
)	O
.	O

Treatment	O
of	O
both	O
HMy2	B-cell_line
and	I-cell_line
GM4672A	I-cell_line
cells	I-cell_line
with	O
Dex	O
for	O
1-4	O
days	O
inhibits	O
extracellular	O
PA	B-protein
activity	O
in	O
a	O
concentration-dependent	O
manner	O
,	O
being	O
half-maximal	O
at	O
approximately	O
1	O
X	O
10	O
(	O
-9	O
)	O
M	O
.	O

Inhibition	O
of	O
PA	B-protein
in	O
both	O
cell	O
types	O
is	O
specific	O
for	O
active	O
glucocorticoids	O
,	O
and	O
this	O
specificity	O
parallels	O
the	O
ability	O
of	O
various	O
steroids	O
to	O
bind	O
to	O
glucocorticoid	B-protein
receptors	I-protein
.	O

HMy2	B-cell_line
cell	I-cell_line
PA	B-protein
is	O
fully	O
suppressible	O
by	O
Dex	O
,	O
whereas	O
up	O
to	O
one	O
third	O
of	O
the	O
activator	O
expressed	O
by	O
GM4672A	B-cell_line
cells	I-cell_line
is	O
resistant	O
to	O
glucocorticoid	O
inhibition	O
.	O

Mixing	O
experiments	O
using	O
a	O
UK	O
standard	O
and	O
conditioned	O
media	O
from	O
Dex-treated	B-cell_line
cells	I-cell_line
suggest	O
an	O
absence	O
of	O
glucocorticoid-inducible	B-protein
inhibitors	I-protein
to	O
UK	B-protein
or	O
plasmin	B-protein
in	O
both	O
cell	O
types	O
.	O

However	O
,	O
conditioned	O
media	O
from	O
Dex-treated	B-cell_line
GM4672A	I-cell_line
cells	I-cell_line
inhibits	O
a	O
portion	O
of	O
the	O
homologous	O
cellular	O
activator	O
in	O
conditioned	O
media	O
from	O
control	O
GM4672A	B-cell_line
cells	I-cell_line
.	O

Thus	O
,	O
low	O
levels	O
of	O
glucocorticoid-inducible	B-protein
inhibitors	I-protein
may	O
contribute	O
to	O
,	O
but	O
can	O
not	O
fully	O
account	O
for	O
,	O
Dex	O
inhibition	O
of	O
GM4672A	B-protein
PA	I-protein
activity	O
.	O

Glucocorticoid-inducible	B-protein
inhibitors	I-protein
in	O
HMy2	B-cell_line
cells	I-cell_line
are	O
either	O
totally	O
absent	O
or	O
are	O
present	O
at	O
undetectable	O
levels	O
.	O

Thus	O
,	O
regulation	O
of	O
UK-like	B-protein
PAs	I-protein
in	O
HMy2	B-cell_line
and	I-cell_line
GM4672A	I-cell_line
cells	I-cell_line
differs	O
with	O
respect	O
to	O
the	O
extent	O
to	O
which	O
glucocorticoids	O
inhibit	O
constitutively	B-protein
expressed	I-protein
activator	I-protein
levels	O
,	O
as	O
well	O
as	O
the	O
possible	O
contribution	O
of	O
glucocorticoid-inducible	B-protein
inhibitors	I-protein
to	O
the	O
regulatory	O
process	O
in	O
GM4672A	B-cell_line
cells	I-cell_line
.	O

-DOCSTART-	O

Characterization	O
of	O
aldosterone	O
binding	O
sites	O
in	O
circulating	B-cell_type
human	I-cell_type
mononuclear	I-cell_type
leukocytes	I-cell_type
.	O

Aldosterone	O
binding	O
sites	O
in	O
human	B-cell_line
mononuclear	I-cell_line
leukocytes	I-cell_line
were	O
characterized	O
after	O
separation	O
of	O
cells	O
from	O
blood	O
by	O
a	O
Percoll	O
gradient	O
.	O

After	O
washing	O
and	O
resuspension	O
in	O
RPMI-1640	O
medium	O
,	O
cells	O
were	O
incubated	O
at	O
37	O
degrees	O
C	O
for	O
1	O
h	O
with	O
different	O
concentrations	O
of	O
[	O
3H	O
]	O
aldosterone	O
plus	O
a	O
100-fold	O
concentration	O
of	O
RU-26988	O
(	O
11	O
alpha	O
,	O
17	O
alpha-dihydroxy-17	O
beta-propynylandrost-1	O
,	O
4	O
,	O
6-trien-3-one	O
)	O
,	O
with	O
or	O
without	O
an	O
excess	O
of	O
unlabeled	O
aldosterone	O
.	O

Aldosterone	O
binds	O
to	O
a	O
single	O
class	O
of	O
receptors	O
with	O
an	O
affinity	O
of	O
2.7	O
+/-	O
0.5	O
nM	O
(	O
means	O
+/-	O
SD	O
,	O
n	O
=	O
14	O
)	O
and	O
a	O
capacity	O
of	O
290	O
+/-	O
108	O
sites/cell	O
(	O
n	O
=	O
14	O
)	O
.	O

The	O
specificity	O
data	O
show	O
a	O
hierarchy	O
of	O
affinity	O
of	O
desoxycorticosterone	O
=	O
corticosterone	O
=	O
aldosterone	O
greater	O
than	O
hydrocortisone	O
greater	O
than	O
dexamethasone	O
.	O

The	O
results	O
indicate	O
that	O
mononuclear	B-cell_type
leukocytes	I-cell_type
could	O
be	O
useful	O
for	O
studying	O
the	O
physiological	O
significance	O
of	O
these	O
mineralocorticoid	B-protein
receptors	I-protein
and	O
their	O
regulation	O
in	O
humans	O
.	O

-DOCSTART-	O

Mineralocorticoid	B-protein
and	I-protein
glucocorticoid	I-protein
receptors	I-protein
in	O
circulating	B-cell_type
mononuclear	I-cell_type
leukocytes	I-cell_type
of	O
patients	O
with	O
primary	O
hyperaldosteronism	O
.	O

Mineralocorticoid	B-protein
and	I-protein
glucocorticoid	I-protein
receptors	I-protein
were	O
measured	O
in	O
circulating	B-cell_type
mononuclear	I-cell_type
leukocytes	I-cell_type
in	O
5	O
patients	O
affected	O
by	O
Conn	O
's	O
syndrome	O
(	O
3	O
cases	O
of	O
bilateral	O
adrenal	O
hyperplasia	O
and	O
2	O
cases	O
of	O
adenoma	O
plus	O
unilateral	O
hyperplasia	O
)	O
.	O

The	O
number	O
of	O
the	O
binding	O
sites	O
per	O
cell	O
resulted	O
significantly	O
lower	O
(	O
189	O
+/-	O
114	O
,	O
mean	O
+/-	O
SD	O
)	O
,	O
as	O
compared	O
with	O
the	O
normal	O
controls	O
(	O
298	O
+/-	O
105	O
)	O
.	O

The	O
affinity	O
of	O
aldosterone	O
for	O
the	O
receptor	O
was	O
found	O
to	O
be	O
not	O
different	O
than	O
that	O
of	O
healthy	O
control	O
subjects	O
.	O

The	O
capacity	O
and	O
the	O
affinity	O
of	O
dexamethasone	O
for	O
glucocorticoid	B-protein
receptors	I-protein
ranged	O
in	O
the	O
normal	O
values	O
.	O

These	O
data	O
suggest	O
a	O
possible	O
down-regulation	O
of	O
mineralocorticoid	B-protein
receptors	I-protein
in	O
humans	O
.	O

-DOCSTART-	O

Short-term	O
and	O
long-term	O
effects	O
of	O
estrogen	O
on	O
lymphoid	O
tissues	O
and	O
lymphoid	B-cell_type
cells	I-cell_type
with	O
some	O
remarks	O
on	O
the	O
significance	O
for	O
carcinogenesis	O
.	O

Estrogens	O
have	O
long	O
been	O
thought	O
to	O
play	O
a	O
role	O
in	O
regulating	O
the	O
immune	O
system	O
.	O

The	O
difference	O
in	O
some	O
types	O
of	O
immune	O
responses	O
between	O
males	O
and	O
females	O
is	O
well-known	O
,	O
as	O
is	O
the	O
pronounced	O
thymic	O
involution	O
induced	O
by	O
exogenous	O
estrogens	O
.	O

Estrogens	O
stimulate	O
some	O
aspects	O
of	O
macrophage	B-cell_type
activity	O
and	O
,	O
depending	O
on	O
dose	O
and	O
mitogen	B-protein
,	O
inhibit	O
or	O
stimulate	O
lymphocyte	O
proliferative	O
response	O
in	O
vitro	O
.	O

Another	O
example	O
is	O
the	O
estrogen	O
effect	O
on	O
the	O
delayed	O
type	O
hypersensitivity	O
response	O
.	O

A	O
broad	O
review	O
is	O
given	O
of	O
such	O
estrogen	O
effects	O
on	O
lymphoid	B-cell_type
tissue	I-cell_type
and	O
immune	O
response	O
.	O

Most	O
of	O
the	O
studies	O
published	O
so	O
far	O
are	O
phenomenological	O
.	O

However	O
,	O
the	O
recent	O
description	O
of	O
estrogen	B-protein
receptors	I-protein
in	O
the	O
thymus	O
and	O
in	O
some	O
lymphocyte	B-cell_line
subpopulations	I-cell_line
,	O
as	O
well	O
as	O
a	O
deeper	O
understanding	O
of	O
regulating	O
factors	O
in	O
the	O
immune	O
system	O
,	O
open	O
the	O
possibility	O
of	O
a	O
more	O
detailed	O
understanding	O
of	O
the	O
estrogen	O
mechanism	O
of	O
interference	O
.	O

Estrogen	O
effects	O
in	O
adults	O
are	O
reversible	O
.	O

After	O
treating	O
neonatal	O
mice	O
with	O
the	O
synthetic	O
estrogen	O
diethylstilbestrol	O
(	O
DES	O
)	O
,	O
disturbances	O
are	O
induced	O
in	O
lymphocyte	B-cell_line
populations	I-cell_line
and	O
lymphocyte	B-cell_type
functions	O
which	O
are	O
permanent	O
and	O
irreversible	O
.	O

Lymphocytes	B-cell_type
from	O
adult	O
,	O
neonatally	O
DES-treated	O
female	O
mice	O
have	O
a	O
reduced	O
mitogen	O
response	O
to	O
ConA	B-protein
and	O
LPS	B-protein
(	O
T	B-protein
and	I-protein
B	I-protein
cell	I-protein
mitogen	I-protein
)	O
and	O
the	O
delayed	O
type	O
hypersensitivity	O
response	O
is	O
depressed	O
.	O

A	O
detailed	O
analysis	O
demonstrated	O
a	O
decreased	O
T	B-cell_type
helper	I-cell_type
cell	I-cell_type
population	O
.	O

The	O
activity	O
of	O
Natural	B-cell_type
Killer	I-cell_type
cells	I-cell_type
is	O
permanently	O
reduced	O
and	O
this	O
functional	O
impairment	O
is	O
related	O
to	O
a	O
decreased	O
number	O
of	O
these	O
cells	O
,	O
in	O
turn	O
determined	O
at	O
the	O
bone	O
marrow	O
level	O
.	O

The	O
same	O
animals	O
have	O
an	O
increased	O
sensitivity	O
to	O
chemical	O
carcinogens	O
(	O
methylcholanthrene	O
)	O
and	O
they	O
spontaneously	O
develop	O
epithelial	O
changes	O
in	O
the	O
uterine	O
cervix	O
which	O
morphologically	O
are	O
similar	O
to	O
adenocarcinoma	O
.	O

The	O
association	O
between	O
estrogen-associated	O
malignancy	O
and	O
estrogen	O
effects	O
in	O
lymphocyte	B-cell_type
functions	O
deserves	O
further	O
study	O
.	O

-DOCSTART-	O

Drugs	O
affecting	O
the	O
hormonal	O
receptors	O
of	O
normal	B-cell_type
and	I-cell_type
leukaemic	I-cell_type
peripheral	I-cell_type
leucocytes	I-cell_type
.	O

The	O
authors	O
investigated	O
the	O
behaviour	O
of	O
steroid	O
hormone	O
uptake	O
in	O
leukaemic	B-cell_type
cells	I-cell_type
(	O
CML	B-cell_line
,	O
CLL	B-cell_line
,	O
AML	B-cell_line
,	O
ALL	B-cell_line
)	O
,	O
in	O
basal	O
conditions	O
and	O
after	O
incubation	O
with	O
drugs	O
which	O
modify	O
the	O
cellular	O
concentration	O
of	O
cAMP	O
,	O
PGE	O
and	O
PGF	O
.	O

The	O
results	O
demonstrated	O
the	O
presence	O
in	O
leukaemic	B-cell_type
cells	I-cell_type
of	O
an	O
alteration	O
in	O
the	O
incorporation	O
of	O
steroid	O
hormones	O
.	O

This	O
alteration	O
was	O
scarcely	O
modified	O
by	O
incubation	O
with	O
theophylline	B-protein
,	O
which	O
increases	O
cellular	O
concentration	O
of	O
cAMP	O
.	O

On	O
the	O
other	O
hand	O
,	O
it	O
was	O
moderately	O
counteracted	O
by	O
thioproline	O
and	O
was	O
evidently	O
inhibited	O
by	O
flurbiprofen	O
,	O
which	O
also	O
reduced	O
cellular	O
concentrations	O
of	O
prostaglandins	O
,	O
particularly	O
PGE2	O
,	O
with	O
the	O
exception	O
of	O
PGF2	O
which	O
showed	O
a	O
poor	O
response	O
.	O

Differences	O
were	O
observed	O
in	O
the	O
behavior	O
of	O
hormonal	O
uptake	O
of	O
CML	B-cell_line
,	O
in	O
contrast	O
to	O
that	O
of	O
AML	B-cell_line
,	I-cell_line
CLL	I-cell_line
and	I-cell_line
ALL	I-cell_line
peripheral	I-cell_line
leucocytes	I-cell_line
.	O

-DOCSTART-	O

Human	O
breast	O
cancer	O
and	O
impaired	O
NK	B-cell_type
cell	I-cell_type
function	O
.	O

Recent	O
advances	O
in	O
tumor	O
immunology	O
have	O
led	O
to	O
the	O
discovery	O
of	O
a	O
new	O
lymphoid	B-cell_type
cell	I-cell_type
with	O
unique	O
antitumor	O
activity	O
.	O

Natural	B-cell_type
killer	I-cell_type
(	I-cell_type
NK	I-cell_type
)	I-cell_type
cells	I-cell_type
form	O
an	O
antitumor	O
surveillance	O
system	O
and	O
appear	O
to	O
be	O
vital	O
in	O
preventing	O
tumor	O
growth	O
and	O
metastasis	O
in	O
animal	O
models	O
.	O

We	O
studied	O
NK	B-cell_type
activity	O
in	O
patients	O
with	O
benign	O
and	O
malignant	O
breast	O
disease	O
,	O
using	O
a	O
chromium-51	O
release	O
microtiter	O
cytotoxicity	O
assay	O
with	O
K562	B-cell_line
cells	I-cell_line
as	O
targets	O
.	O

Compared	O
with	O
benign	O
controls	O
,	O
patients	O
with	O
malignancies	O
had	O
significantly	O
depressed	O
NK	B-cell_type
-mediated	O
lysis	O
(	O
P	O
less	O
than	O
0.01	O
)	O
.	O

Furthermore	O
,	O
lysis	O
in	O
those	O
with	O
advanced	O
disease	O
(	O
stages	O
II	O
,	O
III	O
,	O
and	O
IV	O
)	O
was	O
significantly	O
less	O
than	O
in	O
those	O
with	O
limited	O
disease	O
(	O
stage	O
I	O
)	O
(	O
P	O
less	O
than	O
0.01	O
)	O
.	O

NK	B-cell_type
activity	O
was	O
not	O
correlated	O
to	O
estrogen	O
or	O
progesterone	O
receptor	O
states	O
.	O

Positive	O
correlation	O
of	O
a	O
depressed	O
natural	O
killer	O
activity	O
with	O
the	O
extent	O
of	O
tumor	O
spread	O
supports	O
the	O
concept	O
of	O
an	O
NK	B-cell_type
cell	I-cell_type
immune	O
surveillance	O
system	O
in	O
breast	O
cancer	O
and	O
emphasizes	O
its	O
importance	O
in	O
this	O
malignancy	O
.	O

-DOCSTART-	O

Immunosuppressive	O
effect	O
of	O
serum	O
progesterone	O
during	O
pregnancy	O
depends	O
on	O
the	O
progesterone	O
binding	O
capacity	O
of	O
the	O
lymphocytes	B-cell_type
.	O

Cytotoxic	O
activity	O
and	O
progesterone	O
binding	O
capacity	O
of	O
the	O
lymphocytes	B-cell_type
,	O
together	O
with	O
serum	O
progesterone	O
concentrations	O
,	O
were	O
determined	O
in	O
women	O
with	O
normal	O
pregnancy	O
or	O
with	O
a	O
clinical	O
diagnosis	O
of	O
threatened	O
abortion	O
or	O
threatened	O
premature	O
labour	O
.	O

The	O
lymphocytes	O
of	O
women	O
with	O
threatened	O
abortion	O
or	O
threatened	O
premature	O
labour	O
showed	O
significantly	O
higher	O
cytotoxic	O
activity	O
(	O
P	O
less	O
than	O
0.001	O
)	O
and	O
significantly	O
lower	O
progesterone	O
binding	O
capacity	O
(	O
P	O
less	O
than	O
0.001	O
)	O
than	O
did	O
lymphocytes	B-cell_type
obtained	O
from	O
the	O
healthy	O
pregnant	O
women	O
.	O

Significant	O
inverse	O
correlation	O
was	O
found	O
between	O
progesterone	O
binding	O
capacity	O
and	O
cytotoxic	O
activity	O
of	O
the	O
lymphocytes	B-cell_type
(	O
P	O
less	O
than	O
0.001	O
)	O
,	O
but	O
the	O
progesterone	O
concentration	O
of	O
the	O
pregnancy	O
serum	O
appeared	O
to	O
have	O
no	O
influence	O
on	O
the	O
other	O
two	O
parameters	O
.	O

The	O
findings	O
indicate	O
that	O
intact	O
progesterone	O
binding	O
capacity	O
of	O
the	O
lymphocytes	B-cell_type
is	O
an	O
essential	O
factor	O
for	O
the	O
manifestation	O
of	O
the	O
blocking	O
effect	O
exerted	O
by	O
pregnancy	O
serum	O
on	O
lymphocyte	B-cell_type
cytotoxicity	O
in	O
vitro	O
.	O

-DOCSTART-	O

Serum	O
sex	O
steroid	O
and	O
peptide	O
hormone	O
concentrations	O
,	O
and	O
endometrial	O
estrogen	O
and	O
progestin	O
receptor	O
levels	O
during	O
administration	O
of	O
human	B-protein
leukocyte	I-protein
interferon	I-protein
.	O

Five	O
normally	O
cycling	O
healthy	O
women	O
were	O
given	O
daily	O
subcutaneous	O
injections	O
of	O
human	B-protein
leukocyte	I-protein
interferon	I-protein
(	O
3	O
X	O
10	O
(	O
6	O
)	O
units/day	O
)	O
from	O
the	O
3rd	O
through	O
23rd	O
day	O
of	O
the	O
menstrual	O
cycle	O
,	O
and	O
serum	O
steroid	O
and	O
peptide	O
hormone	O
concentrations	O
monitored	O
at	O
3-day	O
intervals	O
during	O
the	O
treatment	O
and	O
the	O
preceding	O
control	O
cycle	O
.	O

Concentrations	O
of	O
cytosol	B-protein
and	I-protein
nuclear	I-protein
estrogen	I-protein
receptors	I-protein
(	O
ERC	B-protein
and	O
ERN	B-protein
,	O
respectively	O
)	O
and	O
progestin	B-protein
receptors	I-protein
(	O
PRC	B-protein
and	O
PRN	B-protein
)	O
were	O
also	O
measured	O
from	O
endometrial	O
biopsies	O
taken	O
on	O
the	O
24th	O
day	O
of	O
the	O
control	O
and	O
treatment	O
cycle	O
.	O

In	O
addition	O
,	O
an	O
extensive	O
monitoring	O
of	O
clinical	O
chemical	O
and	O
hematological	O
tests	O
from	O
the	O
blood	O
samples	O
were	O
performed	O
.	O

Serum	O
estradiol	O
and	O
progesterone	O
concentrations	O
were	O
significantly	O
decreased	O
during	O
the	O
treatment	O
cycle	O
,	O
suggesting	O
that	O
interferon	B-protein
interacts	O
in	O
vivo	O
with	O
the	O
function	O
of	O
both	O
FSH	O
and	O
LH	O
.	O

No	O
significant	O
changes	O
were	O
observed	O
in	O
the	O
serum	O
peptide	O
hormone	O
concentrations	O
measured	O
(	O
FSH	O
,	O
LH	O
,	O
prolactin	O
,	O
insulin	O
,	O
growth	O
hormone	O
and	O
TSH	O
)	O
;	O
neither	O
were	O
the	O
levels	O
of	O
endometrial	B-protein
ERC	I-protein
,	I-protein
ERN	I-protein
,	I-protein
PRC	I-protein
and	I-protein
PRN	I-protein
affected	O
by	O
interferon	B-protein
administration	O
.	O

As	O
expected	O
,	O
interferon	B-protein
administration	O
resulted	O
in	O
decreased	O
leukocyte	B-cell_type
counts	O
.	O

Moreover	O
,	O
an	O
increasing	O
tendency	O
in	O
the	O
activities	O
of	O
serum	B-protein
alkaline	I-protein
phosphatase	I-protein
and	O
gamma-glutamyltransferase	B-protein
during	O
the	O
interferon	B-protein
therapy	O
shows	O
that	O
interferon	B-protein
may	O
slightly	O
interfere	O
with	O
the	O
liver	O
function	O
.	O

These	O
results	O
suggest	O
that	O
one	O
of	O
the	O
mechanisms	O
by	O
which	O
interferon	B-protein
treatment	O
may	O
affect	O
the	O
growth	O
of	O
hormone-dependent	O
neoplasms	O
could	O
be	O
the	O
interaction	O
with	O
production	O
and/or	O
function	O
of	O
circulating	O
hormonal	O
compounds	O
.	O

-DOCSTART-	O

[	O
Glucocorticoid	B-protein
receptor	I-protein
level	O
in	O
the	O
blood	B-cell_type
leukocytes	I-cell_type
in	O
different	O
acute	O
diseases	O
]	O

Content	O
of	O
glucocorticoid	B-protein
receptors	I-protein
in	O
cytosol	O
of	O
blood	B-cell_type
leukocytes	I-cell_type
,	O
concentration	O
of	O
cortisol	O
and	O
amount	O
of	O
leukocytes	B-cell_type
in	O
blood	O
were	O
studied	O
in	O
20	O
patients	O
with	O
acute	O
impairments	O
within	O
the	O
second	O
day	O
of	O
the	O
disease	O
.	O

Content	O
of	O
receptors	O
in	O
cytosol	O
of	O
blood	O
leukocytes	O
was	O
studied	O
using	O
3H-triamcinolone	O
acetonide	O
.	O

Distinct	O
increase	O
in	O
amount	O
of	O
the	O
leukocyte	B-protein
glucocorticoid	I-protein
receptors	I-protein
was	O
found	O
in	O
patients	O
with	O
poisoning	O
by	O
dichlorethane	O
and	O
hypnotic	O
drugs	O
under	O
conditions	O
of	O
acute	O
myocardial	O
infarction	O
.	O

In	O
acute	O
pancreatitis	O
content	O
of	O
the	O
leukocyte	B-protein
receptors	I-protein
was	O
not	O
altered	O
as	O
compared	O
with	O
controls	O
.	O

Concentration	O
of	O
endogenous	O
cortisol	O
was	O
increased	O
in	O
blood	O
of	O
all	O
the	O
patients	O
,	O
except	O
of	O
the	O
cases	O
of	O
acetate	O
intoxication	O
.	O

Reverse	O
correlation	O
was	O
observed	O
between	O
concentration	O
of	O
cortisol	O
in	O
blood	O
and	O
content	O
of	O
glucocorticoid	O
receptors	O
in	O
leukocytes	B-cell_type
.	O

But	O
in	O
the	O
patients	O
with	O
acute	O
pancreatitis	O
the	O
decrease	O
in	O
content	O
of	O
leukocyte	B-protein
glucocorticoid	I-protein
receptors	I-protein
was	O
not	O
observed	O
although	O
there	O
was	O
an	O
increase	O
in	O
cortisol	O
concentration	O
in	O
blood	O
.	O

The	O
role	O
of	O
glucocorticoid	B-protein
receptors	I-protein
in	O
immunological	O
processes	O
under	O
conditions	O
of	O
purulent	O
complications	O
and	O
possibility	O
to	O
regulate	O
the	O
metabolism	O
in	O
leukocytes	B-cell_type

-DOCSTART-	O

Therapeutic	O
concentrations	O
of	O
glucocorticoids	O
suppress	O
the	O
antimicrobial	O
activity	O
of	O
human	B-cell_type
macrophages	I-cell_type
without	O
impairing	O
their	O
responsiveness	O
to	O
gamma	B-protein
interferon	I-protein
.	O

By	O
exposing	O
human	B-cell_type
blood-derived	I-cell_type
macrophages	I-cell_type
and	I-cell_type
alveolar	I-cell_type
macrophages	I-cell_type
in	O
vitro	O
to	O
dexamethasone	O
,	O
we	O
showed	O
in	O
these	O
studies	O
that	O
glucocorticoids	O
markedly	O
suppress	O
the	O
antimicrobial	O
activity	O
of	O
macrophages	B-cell_type
but	O
not	O
macrophage	B-cell_type
activation	O
by	O
lymphokines	B-cell_type
.	O

As	O
little	O
as	O
2.5	O
X	O
10	O
(	O
-8	O
)	O
mol/liter	O
of	O
dexamethasone	O
prevented	O
macrophages	B-cell_type
from	O
inhibiting	O
germination	O
of	O
Aspergillus	O
spores	O
or	O
from	O
eliminating	O
ingested	O
bacteria	O
such	O
as	O
Listeria	O
,	O
Nocardia	O
,	O
or	O
Salmonella	O
.	O

Damage	O
to	O
macrophage	B-cell_type
function	O
was	O
inhibited	O
by	O
progesterone	O
and	O
appeared	O
to	O
be	O
receptor-mediated	O
.	O

In	O
accordance	O
with	O
in	O
vivo	O
observations	O
,	O
dexamethasone	O
required	O
24-36	O
h	O
to	O
suppress	O
antimicrobial	O
activity	O
.	O

While	O
glucocorticoids	O
interfered	O
with	O
base-line	O
activity	O
of	O
macrophages	B-cell_type
,	O
dexamethasone	O
concentrations	O
comparable	O
to	O
drug	O
levels	O
in	O
patients	O
had	O
no	O
effect	O
on	O
macrophage	B-cell_type
activation	O
.	O

Proliferating	O
lymphocytes	B-cell_type
and	O
gamma-interferon	B-protein
thus	O
increased	O
the	O
antimicrobial	O
activity	O
of	O
phagocytes	B-cell_type
exposed	O
to	O
glucocorticoids	O
over	O
that	O
of	O
control	O
cells	O
.	O

Macrophage	O
activation	O
and	O
correction	O
of	O
the	O
dexamethasone	O
effect	O
by	O
gamma-interferon	B-protein
,	O
however	O
,	O
was	O
dependent	O
on	O
the	O
pathogen	O
.	O

The	O
lymphokine	B-protein
enhanced	O
the	O
antimicrobial	O
activity	O
of	O
dexamethasone-treated	B-cell_line
macrophages	I-cell_line
against	O
Listeria	O
and	O
Salmonella	O
but	O
not	O
against	O
Aspergillus	O
or	O
Nocardia	O
.	O

Dexamethasone-induced	O
damage	O
to	O
the	O
antimicrobial	O
activity	O
of	O
human	B-cell_type
macrophages	I-cell_type
in	O
vitro	O
parallels	O
observations	O
that	O
glucocorticoids	O
render	O
laboratory	O
animals	O
susceptible	O
to	O
listeriosis	O
and	O
aspergillosis	O
by	O
damaging	O
resident	O
macrophages	B-cell_line
.	O

Suppression	O
of	O
macrophage	B-cell_type
antimicrobial	O
activity	O
should	O
thus	O
be	O
considered	O
when	O
treating	O
patients	O
with	O
glucocorticoids	O
;	O
its	O
prevention	O
by	O
gamma-interferon	B-protein
might	O
be	O
beneficial	O
for	O
some	O
but	O
not	O
all	O
pathogens	O
.	O

-DOCSTART-	O

Interleukin	B-protein
2	I-protein
receptor	I-protein
(	O
Tac	B-protein
antigen	I-protein
)	O
expression	O
in	O
HTLV-I-associated	O
adult	O
T-cell	B-cell_type
leukemia	O
.	O

Interleukin-2	B-protein
(	O
IL-2	B-protein
)	O
is	O
a	O
lymphokine	B-protein
synthesized	O
by	O
some	O
T-cells	B-cell_type
following	O
activation	O
.	O

Resting	O
T-cells	B-cell_type
do	O
not	O
express	O
IL-2	B-protein
receptors	I-protein
,	O
but	O
receptors	O
are	O
rapidly	O
expressed	O
on	O
T-cells	B-cell_type
following	O
interaction	O
of	O
antigens	O
,	O
mitogens	O
,	O
or	O
monoclonal	B-protein
antibodies	I-protein
with	O
the	O
antigen-specific	B-protein
T-cell	I-protein
receptor	I-protein
complex	I-protein
.	O

Using	O
anti-Tac	B-protein
,	O
a	O
monoclonal	B-protein
antibody	I-protein
that	O
recognizes	O
the	O
IL-2	B-protein
receptor	I-protein
,	O
the	O
receptor	O
has	O
been	O
purified	O
and	O
shown	O
to	O
be	O
a	O
Mr	O
33	O
,	O
000	O
peptide	O
that	O
is	O
posttranslationally	O
glycosylated	O
to	O
a	O
Mr	O
55	O
,	O
000	O
mature	O
form	O
.	O

Normal	O
resting	O
T-cells	B-cell_type
and	O
most	O
leukemic	B-cell_line
T-cell	I-cell_line
populations	I-cell_line
do	O
not	O
express	O
IL-2	B-protein
receptors	I-protein
;	O
however	O
,	O
the	O
leukemic	B-cell_type
cells	I-cell_type
of	O
the	O
11	O
patients	O
examined	O
who	O
had	O
human	O
T-cell	O
lymphotropic	O
virus-associated	O
adult	O
T-cell	O
leukemia	O
expressed	O
the	O
Tac	B-protein
antigen	I-protein
.	O

In	O
human	B-cell_type
T-cell	I-cell_type
lymphotropic	I-cell_type
virus-I	I-cell_type
infected	I-cell_type
cells	I-cell_type
,	O
the	O
Mr	O
42	O
,	O
000	O
long	O
open	B-protein
reading	I-protein
frame	I-protein
protein	I-protein
encoded	O
in	O
part	O
by	O
the	O
pX	B-DNA
region	I-DNA
of	O
this	O
virus	O
may	O
act	O
as	O
a	O
transacting	B-protein
transcriptional	I-protein
activator	I-protein
that	O
induces	O
IL-2	B-DNA
receptor	I-DNA
gene	I-DNA
transcription	O
,	O
thus	O
providing	O
an	O
explanation	O
for	O
the	O
constant	O
association	O
of	O
IL-2	B-protein
receptor	I-protein
expression	O
with	O
adult	O
T-cell	O
lymphotropic	O
virus-I	O
infection	O
of	O
lymphoid	B-cell_type
cells	I-cell_type
.	O

The	O
constant	O
expression	O
of	O
large	O
numbers	O
of	O
IL-2	B-protein
receptors	I-protein
which	O
may	O
be	O
aberrant	O
may	O
play	O
a	O
role	O
in	O
the	O
uncontrolled	O
growth	O
of	O
adult	B-cell_type
T-cell	I-cell_type
leukemia	I-cell_type
cells	I-cell_type
.	O

Two	O
patients	O
with	O
Tac-positive	O
adult	O
T-cell	O
leukemia	O
have	O
been	O
treated	O
with	O
the	O
anti-Tac	B-protein
.	O

One	O
of	O
the	O
patients	O
had	O
6-	O
and	O
3-mo	O
remissions	O
of	O
his	O
leukemia	O
following	O
two	O
courses	O
of	O
therapy	O
with	O
this	O
monoclonal	B-protein
antibody	I-protein
directed	O
toward	O
this	O
growth	B-protein
factor	I-protein
receptor	I-protein
.	O

-DOCSTART-	O

Lymphocyte	B-cell_type
glucocorticoid	B-protein
receptor	I-protein
binding	O
in	O
depressed	O
patients	O
with	O
hypercortisolemia	O
.	O

Despite	O
elevated	O
levels	O
of	O
serum	O
and	O
urinary	O
cortisol	O
,	O
patients	O
with	O
depressive	O
illness	O
manifest	O
none	O
of	O
the	O
clinical	O
stigmata	O
of	O
glucocorticoid	O
excess	O
.	O

This	O
hypercortisolemia	O
in	O
the	O
absence	O
of	O
clinical	O
effects	O
suggests	O
a	O
state	O
of	O
hormone	O
resistance	O
and	O
could	O
be	O
mediated	O
by	O
alterations	O
in	O
the	O
glucocorticoid	B-protein
receptor	I-protein
.	O

Earlier	O
studies	O
have	O
shown	O
that	O
small	O
doses	O
of	O
glucocorticoids	O
cause	O
a	O
decrease	O
in	O
glucocorticoid	B-protein
receptor	I-protein
binding	O
in	O
normal	O
human	B-cell_type
lymphocytes	I-cell_type
.	O

White	O
cells	O
from	O
depressed	O
patients	O
with	O
significant	O
hypercortisolemia	O
would	O
be	O
expected	O
to	O
show	O
a	O
similar	O
change	O
in	O
receptor	O
concentration	O
if	O
peripheral	O
tissues	O
are	O
adequately	O
exposed	O
to	O
and	O
sensitive	O
to	O
the	O
hormone	O
.	O

In	O
this	O
study	O
we	O
compared	O
the	O
binding	O
of	O
[	O
3H	O
]	O
dexamethasone	O
to	O
lymphocytes	B-cell_type
from	O
normal	O
subjects	O
and	O
depressed	O
patients	O
with	O
hypercortisolemia	O
.	O

Lymphocytes	B-cell_type
from	O
normal	O
subjects	O
had	O
a	O
mean	O
receptor	O
concentration	O
of	O
10.2	O
+/-	O
0.66	O
fm/10	O
(	O
6	O
)	O
cells	O
(	O
S.E.M.	O
)	O
and	O
a	O
dissociation	O
constant	O
of	O
4.8	O
+/-	O
0.47	O
nM	O
.	O

Lymphocytes	B-cell_type
from	O
depressed	O
patients	O
with	O
abnormal	O
0800	O
h	O
serum	O
cortisol	O
after	O
dexamethasone	O
had	O
a	O
mean	O
receptor	O
concentration	O
of	O
8.8	O
+/-	O
0.75	O
fm/10	O
(	O
6	O
)	O
cells	O
,	O
which	O
was	O
not	O
significantly	O
different	O
from	O
that	O
in	O
lymphocytes	B-cell_type
from	O
normal	O
subjects	O
or	O
from	O
depressed	O
subjects	O
with	O
normal	O
post-dexamethasone	O
cortisol	O
levels	O
(	O
9.4	O
+/-	O
0.95	O
fm/10	O
(	O
6	O
)	O
cells	O
)	O
.	O

Lymphocytes	B-cell_type
from	O
depressed	O
patients	O
with	O
elevated	O
urinary	O
free	O
cortisol	O
excretion	O
(	O
UFC	O
)	O
also	O
had	O
normal	O
receptor	O
concentration	O
and	O
binding	O
affinity	O
for	O
dexamethasone	O
.	O

The	O
lack	O
of	O
a	O
change	O
in	O
lymphocyte	B-cell_type
glucocorticoid	B-protein
receptor	I-protein
concentration	O
in	O
the	O
presence	O
of	O
cortisol	O
excess	O
suggests	O
the	O
possibility	O
that	O
hypercortisolemia	O
in	O
depressive	O
illness	O
represents	O
a	O
state	O
of	O
peripheral	O
glucocorticoid	O
resistance	O
.	O

-DOCSTART-	O

Structure	O
and	O
regulation	O
of	O
the	O
glucocorticoid	B-protein
hormone	I-protein
receptor	I-protein
.	O

The	O
glucocorticoid	B-protein
receptor	I-protein
is	O
an	O
intracellular	B-protein
protein	I-protein
which	O
possesses	O
three	O
distinct	O
domains	O
,	O
one	O
that	O
binds	O
agonist	O
and	O
antagonist	O
steroids	O
,	O
one	O
that	O
binds	O
DNA	O
,	O
and	O
one	O
that	O
binds	O
anti-receptor	B-protein
antibodies	I-protein
and	O
is	O
required	O
for	O
glucocorticoid	O
modulation	O
of	O
gene	O
expression	O
.	O

In	O
intact	O
cells	O
,	O
receptor	O
number	O
,	O
affinity	O
and	O
activity	O
can	O
change	O
in	O
response	O
to	O
factors	O
that	O
bind	O
to	O
the	O
receptor	O
,	O
or	O
that	O
act	O
indirectly	O
through	O
ill-defined	O
mechanisms	O
which	O
may	O
include	O
resumption	O
or	O
arrest	O
of	O
cell	O
cycling	O
and	O
variations	O
in	O
intracellular	O
calcium	O
ion	O
concentrations	O
.	O

Some	O
of	O
these	O
factors	O
appear	O
to	O
exert	O
their	O
effect	O
by	O
controlling	O
critical	O
receptor	O
properties	O
such	O
as	O
ATP-dependent	O
phosphorylation	O
,	O
integrity	O
of	O
thiol	B-protein
groups	I-protein
,	O
and	O
exposure	O
of	O
key	B-protein
amino	I-protein
acid	I-protein
residues	I-protein
.	O

Glucocorticoid	O
agonists	O
promote	O
the	O
'transformation	O
'	O
of	O
the	O
receptor	O
into	O
the	O
DNA-binding	O
state	O
,	O
which	O
is	O
competent	O
for	O
modulating	O
gene	O
expression	O
.	O

Glucocorticoid	O
antagonists	O
are	O
steroids	O
that	O
interact	O
with	O
the	O
receptor	O
but	O
either	O
fail	O
to	O
produce	O
a	O
stable	B-protein
complex	I-protein
or	O
produce	O
a	O
stable	B-protein
but	I-protein
inefficient	I-protein
complex	I-protein
.	O

Although	O
substituent	O
groups	O
that	O
confer	O
agonist	O
or	O
antagonist	O
activity	O
to	O
the	O
steroid	O
have	O
been	O
identified	O
,	O
the	O
molecular	O
determinants	O
of	O
this	O
difference	O
at	O
the	O
receptor	O
level	O
remain	O
unknown	O
.	O

Most	O
in	O
vitro	O
and	O
in	O
vivo	O
data	O
on	O
receptor	O
regulation	O
can	O
be	O
accommodated	O
by	O
postulating	O
the	O
existence	O
of	O
an	O
intracellular	O
cycle	O
that	O
involves	O
five	O
states	O
of	O
the	O
receptor	O
.	O

The	O
active	O
free	O
receptor	O
is	O
phosphorylated	O
,	O
reduced	O
,	O
and	O
presumably	O
oligomeric	O
(	O
state	O
A	O
)	O
.	O

Following	O
binding	O
of	O
an	O
agonist	O
(	O
state	O
B	O
)	O
,	O
it	O
can	O
become	O
transformed	O
by	O
dissociation	O
into	O
its	O
subunits	O
and	O
dephosphorylation	O
(	O
state	O
C	O
)	O
.	O

The	O
transformed	B-protein
receptor	I-protein
then	O
interacts	O
with	O
chromatin	B-DNA
(	O
state	O
D	O
)	O
.	O

Dissociation	O
of	O
the	O
steroid	O
and	O
oxidation	O
of	O
receptor	B-protein
thiol	I-protein
group	I-protein
(	O
s	O
)	O
lead	O
to	O
the	O
inactive	O
receptor	O
form	O
(	O
state	O
E	O
)	O
.	O

Reduction	O
and	O
rephosphorylation	O
of	O
the	O
receptor	O
enable	O
it	O
to	O
bind	O
steroids	O
again	O
so	O
that	O
the	O
cycle	O
is	O
closed	O
.	O

-DOCSTART-	O

Thermodynamics	O
of	O
steroid	O
binding	O
to	O
the	O
human	B-protein
glucocorticoid	I-protein
receptor	I-protein
.	O

The	O
thermodynamics	O
of	O
the	O
interaction	O
of	O
glucocorticoids	O
with	O
their	O
receptor	O
were	O
studied	O
in	O
cytosol	O
from	O
human	B-cell_type
lymphoblastoid	I-cell_type
cells	I-cell_type
.	O

The	O
rate	O
and	O
affinity	O
constants	O
of	O
dexamethasone	O
and	O
cortisol	O
between	O
0	O
degree	O
and	O
25	O
degrees	O
C	O
were	O
calculated	O
by	O
curve-fitting	O
from	O
time-course	O
and	O
equilibrium	O
kinetics	O
.	O

The	O
data	O
were	O
consistent	O
with	O
a	O
simple	O
reversible	O
bimolecular	O
interaction	O
.	O

Arrhenius	O
and	O
Va	O
n't	O
Hoff	O
plots	O
were	O
curvilinear	O
for	O
both	O
steroids	O
.	O

At	O
equilibrium	O
,	O
the	O
solution	O
for	O
the	O
equation	O
delta	O
G	O
=	O
delta	O
H	O
-	O
T	O
X	O
delta	O
S	O
(	O
eqn.	O
1	O
)	O
was	O
(	O
in	O
kJ	O
X	O
mol-1	O
)	O
-47	O
=	O
36	O
-	O
83	O
(	O
dexamethasone	O
)	O
and	O
-42	O
=	O
-9	O
-	O
33	O
(	O
cortisol	O
)	O
at	O
0	O
degree	O
C	O
.	O

Enthalpy	O
and	O
entropy	O
changes	O
decreased	O
quasi-linearly	O
with	O
temperature	O
such	O
that	O
,	O
at	O
25	O
degrees	O
C	O
,	O
the	O
respective	O
values	O
were	O
-50	O
=	O
-75	O
+	O
25	O
and	O
-43	O
=	O
-48	O
+	O
5	O
.	O

Thus	O
,	O
for	O
both	O
steroids	O
,	O
the	O
interaction	O
was	O
entropy-driven	O
at	O
low	O
temperature	O
and	O
became	O
entirely	O
enthalpy-driven	O
at	O
20	O
degrees	O
C	O
.	O

Thermodynamic	O
values	O
for	O
the	O
transition	O
state	O
were	O
calculated	O
from	O
the	O
rate	O
constants	O
.	O

For	O
the	O
forward	O
reaction	O
,	O
eqn.	O
(	O
1	O
)	O
gave	O
45	O
=	O
84	O
-	O
39	O
(	O
dexamethasone	O
)	O
and	O
46	O
=	O
60	O
-	O
14	O
(	O
cortisol	O
)	O
at	O
0	O
degree	O
C	O
,	O
and	O
44	O
=	O
24	O
+	O
20	O
(	O
dexamethasone	O
)	O
and	O
46	O
=	O
28	O
+	O
18	O
(	O
cortisol	O
)	O
at	O
25	O
degrees	O
C	O
.	O

These	O
data	O
fit	O
quite	O
well	O
with	O
a	O
two-step	O
model	O
[	O
Ross	O
&	O
Subramanian	O
(	O
1981	O
)	O
Biochemistry	O
20	O
,	O
3096-3102	O
]	O
proposed	O
for	O
ligand-protein	O
interactions	O
,	O
which	O
involves	O
a	O
partial	O
immobilization	O
of	O
the	O
reacting	O
species	O
governed	O
by	O
hydrophobic	O
forces	O
,	O
followed	O
by	O
stabilization	O
of	O
the	O
complex	O
by	O
short-range	O
interactions	O
.	O

On	O
the	O
basis	O
of	O
this	O
model	O
,	O
an	O
analysis	O
of	O
the	O
transition-state	O
thermodynamics	O
led	O
to	O
the	O
conclusion	O
that	O
no	O
more	O
than	O
half	O
of	O
the	O
steroid	O
molecular	O
area	O
is	O
engaged	O
in	O
the	O
binding	O
process	O
.	O

-DOCSTART-	O

Cell	O
cycle-related	O
changes	O
in	O
number	O
of	O
T-lymphocyte	B-protein
receptors	I-protein
for	O
glucocorticoids	O
and	O
insulin	O
.	O

Enriched	O
human	B-cell_type
peripheral	I-cell_type
T-lymphocytes	I-cell_type
were	O
stimulated	O
with	O
PHA	O
and	O
examined	O
for	O
variations	O
in	O
insulin	O
and	O
glucocorticoid	O
(	O
dexamethasone	O
)	O
receptor	O
numbers	O
during	O
the	O
early	O
phases	O
of	O
the	O
cell	O
cycle	O
.	O

Cells	O
in	O
G0	O
,	O
G1a	O
and	O
G1b	O
phases	O
,	O
where	O
the	O
G1a	O
-	O
G1b	O
transition	O
is	O
an	O
Interleukin	B-protein
2	I-protein
dependent	O
event	O
,	O
were	O
quantitated	O
by	O
flow	O
cytometry	O
.	O

Few	O
but	O
significant	O
numbers	O
of	O
glucocorticoid	B-protein
receptors	I-protein
(	O
2700/cell	O
)	O
and	O
no	O
insulin	B-protein
receptors	I-protein
(	O
-1/cell	O
)	O
were	O
found	O
in	O
the	O
resting	O
(	O
G0	O
)	O
phase	O
.	O

As	O
cells	O
entered	O
the	O
G1a	O
phase	O
the	O
specific	O
binding	O
of	O
dexamethasone	O
increased	O
and	O
of	O
insulin	O
took	O
place	O
.	O

Although	O
the	O
specific	O
binding	O
further	O
increased	O
as	O
T-cells	B-cell_type
entered	O
the	O
G1b	O
phase	O
(	O
as	O
measured	O
at	O
44	O
h	O
of	O
incubation	O
and	O
using	O
hydroxyurea-treated	B-cell_line
cells	I-cell_line
)	O
,	O
the	O
major	O
changes	O
in	O
the	O
specific	O
binding	O
of	O
dexamethasone	O
took	O
place	O
during	O
the	O
period	O
16	O
-	O
20	O
h	O
after	O
stimulation	O
.	O

Based	O
on	O
these	O
findings	O
,	O
it	O
is	O
concluded	O
that	O
both	O
receptor	O
types	O
(	O
cell	B-protein
membrane	I-protein
and	I-protein
cytoplasmic	I-protein
receptors	I-protein
)	O
are	O
being	O
formed	O
and	O
increased	O
at	O
G1	O
phase	O
prior	O
to	O
cell	O
proliferation	O
,	O
indicating	O
the	O
importance	O
of	O
G1	O
phase	O
in	O
immunoregulation	O
.	O

-DOCSTART-	O

Glucocorticoid	B-protein
receptors	I-protein
and	O
cortico-sensitivity	O
in	O
a	O
human	B-cell_line
clonal	I-cell_line
monocytic	I-cell_line
cell	I-cell_line
line	I-cell_line
,	O
CM-SM	B-cell_line
.	O

CM-SM	B-cell_line
is	O
a	O
clonal	B-cell_line
line	I-cell_line
of	O
human	B-cell_type
precursor	I-cell_type
mononuclear	I-cell_type
phagocytes	I-cell_type
inducible	O
to	O
macrophage	B-cell_type
differentiation	O
in	O
response	O
to	O
the	O
tumor	O
promoter	O
phorbol	O
ester	O
12-O-tetradecanoyl-phorbol-13-acetate	O
(	O
TPA	O
)	O
.	O

Untreated	O
CM-SM	O
cells	O
contain	O
single	O
class	O
,	O
high-affinity	O
(	O
KD	O
=	O
4.0	O
X	O
10	O
(	O
-9	O
)	O
M	O
)	O
glucocorticoid-specific	O
receptor	O
sites	O
(	O
approximately	O
60	O
,	O
000	O
per	O
cell	O
)	O
,	O
as	O
measured	O
by	O
a	O
whole	O
cell	O
assay	O
,	O
at	O
37	O
degrees	O
C	O
,	O
using	O
[	O
3H	O
]	O
triamcinolone	O
acetonide	O
(	O
TA	O
)	O
.	O

Exposure	O
of	O
CM-SM	B-cell_line
to	O
dexamethasone	O
(	O
DEX	O
)	O
produced	O
a	O
progressive	O
,	O
dose-	O
and	O
time-related	O
series	O
of	O
changes	O
in	O
CM-SM	B-cell_line
cell	O
growth	O
,	O
saturation	O
density	O
,	O
morphology	O
,	O
and	O
functional	O
properties	O
,	O
with	O
half-maximal	O
effects	O
at	O
about	O
10	O
(	O
-9	O
)	O
M	O
for	O
DEX	O
.	O

TA-receptor	O
sites	O
rapidly	O
decreased	O
(	O
about	O
70	O
%	O
)	O
after	O
DEX	O
treatment	O
,	O
without	O
any	O
apparent	O
change	O
in	O
steroid	O
specificity	O
and	O
affinity	O
.	O

After	O
5	O
days	O
in	O
culture	O
with	O
a	O
saturating	O
concentration	O
(	O
3.6	O
X	O
10	O
(	O
-8	O
)	O
M	O
)	O
of	O
hormone	O
,	O
the	O
cells	O
reached	O
a	O
saturation	O
density	O
of	O
about	O
9.0	O
X	O
10	O
(	O
6	O
)	O
viable	O
cells/ml	O
(	O
about	O
4.0	O
X	O
10	O
(	O
6	O
)	O
viable	O
cells/ml	O
in	O
the	O
controls	O
)	O
,	O
while	O
the	O
modal	O
volume	O
of	O
the	O
resulting	O
cell	B-cell_line
population	I-cell_line
was	O
approximately	O
60	O
%	O
,	O
as	O
compared	O
to	O
the	O
volume	O
of	O
untreated	B-cell_line
cells	I-cell_line
.	O

DEX-treated	B-cell_line
cells	I-cell_line
appeared	O
less	O
differentiated	O
than	O
controls	O
,	O
as	O
assessed	O
by	O
combined	O
morphologic	O
,	O
antigenic	O
,	O
and	O
cytoenzymatic	O
analyses	O
.	O

DEX	O
almost	O
completely	O
inhibited	O
TPA	O
activation	O
of	O
the	O
following	O
macrophage	B-cell_type
functions	O
:	O
adherency	O
to	O
the	O
culture	O
plate	O
,	O
expression	O
of	O
lysosomal	B-protein
enzymes	I-protein
,	O
Fc	B-protein
and	I-protein
C3	I-protein
receptors	I-protein
,	O
and	O
stimulation	O
of	O
phagocytosis	O
.	O

After	O
removal	O
of	O
DEX	O
,	O
the	O
cells	O
,	O
within	O
a	O
few	O
passages	O
,	O
returned	O
to	O
a	O
state	O
apparently	O
identical	O
to	O
the	O
untreated	O
controls	O
and	O
could	O
be	O
induced	O
to	O
macrophage	B-cell_type
differentiation	O
in	O
response	O
to	O
TPA	O
.	O

-DOCSTART-	O

Acute	O
lymphoblastic	O
leukemia	O
in	O
children	O
:	O
current	O
status	O
,	O
controversies	O
,	O
and	O
future	O
perspective	O
.	O

Disease-free	O
survival	O
(	O
DFS	O
)	O
in	O
childhood	O
ALL	O
is	O
60	O
%	O
,	O
and	O
survival	O
in	O
good	O
,	O
average	O
,	O
and	O
poor	O
prognostic	O
groups	O
defined	O
by	O
initial	O
WBC	O
and	O
age	O
is	O
90	O
,	O
60	O
,	O
and	O
45	O
%	O
,	O
respectively	O
.	O

Additional	O
immunological	O
,	O
morphological	O
,	O
biochemical	O
,	O
cytokinetic	O
,	O
and	O
cytogenetic	O
factors	O
have	O
been	O
identified	O
,	O
illustrating	O
the	O
heterogeneity	O
of	O
ALL	O
and	O
its	O
derivation	O
from	O
malignant	B-cell_line
clones	I-cell_line
at	O
various	O
stages	O
of	O
differentiation	O
and	O
with	O
varying	O
rates	O
of	O
proliferation	O
.	O

Of	O
biologic	O
importance	O
,	O
these	O
factors	O
may	O
refine	O
further	O
the	O
characteristic	O
features	O
of	O
clinically-determined	O
prognostic	O
groups	O
.	O

Multivariate	O
analysis	O
of	O
large	O
prospective	O
trials	O
with	O
homogeneous	O
therapy	O
will	O
be	O
required	O
to	O
determine	O
the	O
independent	O
prognostic	O
importance	O
of	O
these	O
factors	O
.	O

Current	O
treatment	O
strategies	O
in	O
ALL	O
include	O
(	O
1	O
)	O
tailoring	O
therapy	O
and	O
its	O
intensity	O
to	O
prognostic	O
groups	O
;	O
(	O
2	O
)	O
multiple-drug	O
combinations	O
in	O
induction	O
;	O
(	O
3	O
)	O
early	O
use	O
of	O
intrathecal	O
(	O
IT	O
)	O
methotrexate	O
(	O
MTX	O
)	O
;	O
(	O
4	O
)	O
CNS	O
prophylaxis	O
with	O
IT	O
MTX	O
alone	O
in	O
good	O
prognosis	O
patients	O
and	O
combined	O
cranial	O
radiation	O
(	O
CXRT	O
)	O
,	O
1800	O
rads	O
plus	O
IT	O
MTX	O
,	O
in	O
average	O
and	O
poor	O
prognosis	O
patients	O
.	O

Current	O
studies	O
show	O
a	O
CNS	O
relapse	O
rate	O
of	O
5	O
%	O
in	O
all	O
prognostic	O
groups	O
.	O

Late	O
neuropsychological	O
defects	O
caused	O
by	O
cranial	O
XRT	O
and	O
IT	O
MTX	O
have	O
prompted	O
programs	O
designed	O
to	O
reduce	O
the	O
potential	O
late	O
toxicity	O
of	O
CNS	O
prophylaxis	O
.	O

More	O
pronounced	O
in	O
younger	O
children	O
,	O
these	O
abnormalities	O
include	O
decreased	O
IQ	O
,	O
visual-motor	O
incoordination	O
,	O
poor	O
performance	O
in	O
mathematics	O
,	O
and	O
memory	O
dysfunction	O
.	O

Until	O
1980	O
,	O
more	O
intensive	O
induction	O
,	O
consolidation	O
,	O
and	O
maintenance	O
therapy	O
had	O
failed	O
to	O
prolong	O
DFS	O
in	O
children	O
with	O
a	O
poor	O
prognosis	O
.	O

In	O
West	O
Germany	O
(	O
Berlin-Frankfurt-Muenster	O
protocol	O
)	O
a	O
70	O
to	O
75	O
%	O
DFS	O
is	O
seen	O
in	O
all	O
patients	O
regardless	O
of	O
initial	O
WBC	O
,	O
suggesting	O
that	O
effective	O
therapy	O
will	O
override	O
prognostic	O
factors	O
.	O

Ultra-high-dose	O
MTX	O
,	O
without	O
cranial	O
radiation	O
,	O
is	O
also	O
showing	O
promise	O
in	O
poor	O
prognosis	O
patients	O
.	O

Other	O
issues	O
include	O
the	O
optimal	O
duration	O
of	O
therapy	O
,	O
the	O
role	O
of	O
testicular	O
biopsies	O
,	O
and	O
prophylactic	O
testicular	O
radiation	O
.	O

Recent	O
studies	O
suggest	O
that	O
prognostic	O
factors	O
lose	O
their	O
significance	O
after	O
2	O
years	O
of	O
continuous	O
complete	O
remission	O
and	O
that	O
2	O
years	O
of	O
maintenance	O
therapy	O
is	O
adequate	O
.	O

Bilateral	O
open-wedge	O
testicular	O
biopsies	O
have	O
identified	O
occult	O
testicular	O
disease	O
in	O
8	O
to	O
10	O
%	O
of	O
males	O
.	O

A	O
unified	O
approach	O
to	O
children	O
with	O
leukemia/lymphoma	O
,	O
a	O
group	O
with	O
a	O
particularly	O
poor	O
prognosis	O
,	O
utilizing	O
NHL-type	O
therapy	O
may	O
be	O
more	O
effective	O
than	O
conventional	O
ALL	O
therapy	O
.	O

(	O
ABSTRACT	O
TRUNCATED	O
AT	O
400	O
WORDS	O
)	O

-DOCSTART-	O

Glucocorticoid	B-protein
receptor	I-protein
and	O
in	O
vitro	O
sensitivity	O
to	O
steroid	O
hormones	O
in	O
human	O
lymphoproliferative	O
diseases	O
and	O
myeloid	O
leukemia	O
.	O

The	O
glucocorticoid	B-protein
receptor	I-protein
(	O
GR	B-protein
)	O
quantitation	O
by	O
a	O
whole-cell	O
assay	O
and/or	O
cytosol	O
technique	O
and	O
the	O
in	O
vitro	O
sensitivity	O
to	O
steroids	O
have	O
been	O
assessed	O
in	O
peripheral	B-cell_type
blood	I-cell_type
cells	I-cell_type
from	O
normal	O
donors	O
and	O
patients	O
with	O
chronic	O
lymphatic	O
leukemia	O
(	O
CLL	O
)	O
,	O
acute	O
lymphoblastic	O
leukemia	O
(	O
ALL	O
)	O
,	O
lymphosarcoma	O
cell	O
leukemia	O
(	O
LSCL	O
)	O
,	O
acute	O
nonlymphatic	O
leukemia	O
(	O
ANLL	O
)	O
,	O
and	O
chronic	O
myeloid	O
leukemia	O
(	O
CML	O
)	O
.	O

Within	O
the	O
lymphoproliferative	O
diseases	O
,	O
ALL	B-cell_line
cells	I-cell_line
exhibited	O
the	O
highest	O
GR	B-protein
concentration	O
(	O
regardless	O
of	O
the	O
method	O
used	O
)	O
and	O
the	O
highest	O
in	O
vitro	O
inhibition	O
of	O
spontaneous	O
[	O
3H	O
]	O
thymidine	O
(	O
[	O
3H	O
]	O
TdR	O
)	O
uptake	O
by	O
glucocorticoids	O
.	O

A	O
significant	O
relationship	O
between	O
GR	B-protein
concentration	O
(	O
whole-cell	O
assay	O
)	O
and	O
in	O
vitro	O
sensitivity	O
to	O
dexamethasone	O
was	O
also	O
found	O
.	O

On	O
the	O
contrary	O
,	O
CLL	B-cell_line
cells	I-cell_line
presented	O
the	O
highest	O
sensitivity	O
to	O
glucocorticoids	O
in	O
PHA-stimulated	B-cell_line
cell	I-cell_line
cultures	I-cell_line
.	O

Cells	O
from	O
the	O
only	O
two	O
ALL	O
patients	O
who	O
did	O
not	O
undergo	O
a	O
remission	O
after	O
glucocorticoid-inclusive	O
chemotherapy	O
had	O
both	O
the	O
lowest	O
in	O
vitro	O
sensitivity	O
to	O
dexamethasone	O
and	O
the	O
lowest	O
GR	O
concentration	O
with	O
whole-cell	O
assay	O
.	O

Concerning	O
myeloid	O
leukemia	O
,	O
ANLL	O
patients	O
had	O
GR	B-protein
concentrations	O
slightly	O
higher	O
than	O
those	O
found	O
in	O
the	O
ALL	O
group	O
but	O
exhibited	O
the	O
lowest	O
degree	O
of	O
inhibition	O
of	O
spontaneous	O
[	O
3H	O
]	O
TdR	O
uptake	O
by	O
dexamethasone	O
(	O
stimulatory	O
effects	O
occurred	O
in	O
some	O
cases	O
)	O
.	O

CML	B-cell_line
cells	I-cell_line
exhibited	O
an	O
inhibition	O
degree	O
by	O
in	O
vitro	O
glucocorticoids	O
significantly	O
higher	O
than	O
that	O
of	O
ANLL	B-cell_line
cells	I-cell_line
but	O
not	O
different	O
from	O
that	O
of	O
lymphoproliferative	O
diseases	O
.	O

No	O
clear	O
relationship	O
among	O
GR	O
pattern	O
,	O
in	O
vitro	O
cell	O
sensitivity	O
to	O
glucocorticoids	O
,	O
and	O
clinicohematologic	O
parameters	O
was	O
observed	O
in	O
myeloid	O
leukemia-bearing	O
patients	O
.	O

-DOCSTART-	O

Glucocorticoid	B-protein
receptors	I-protein
and	O
in	O
vitro	O
corticosensitivity	O
of	O
peanut-positive	B-cell_line
and	I-cell_line
peanut-negative	I-cell_line
human	I-cell_line
thymocyte	I-cell_line
subpopulations	I-cell_line
.	O

In	O
6	O
human	O
thymus	O
glands	O
,	O
the	O
immature	O
subset	O
of	O
thymocytes	B-cell_type
was	O
separated	O
from	O
the	O
more	O
mature	O
one	O
,	O
by	O
differential	O
peanut	O
lectin	O
agglutination	O
.	O

These	O
2	O
cell	O
subpopulations	O
were	O
analyzed	O
for	O
glucocorticoid	B-protein
receptor	I-protein
content	O
by	O
using	O
a	O
whole	O
cell	O
assay	O
,	O
with	O
(	O
3H	O
)	O
-triamcinolone	O
acetonide	O
as	O
tracer	O
.	O

The	O
unagglutinated	B-cell_type
thymocytes	I-cell_type
(	O
peanut	O
negative	O
)	O
contained	O
about	O
2	O
times	O
more	O
receptor	O
sites	O
per	O
cell	O
than	O
agglutinated	O
(	O
peanut	O
positive	O
)	O
ones	O
(	O
7650	O
+/-	O
1550	O
S.D.	O
verus	O
3195	O
+/-	O
896	O
S.D.	O
)	O
.	O

The	O
affinity	O
for	O
steroid	O
was	O
similar	O
in	O
both	O
cell	O
subsets	O
,	O
as	O
was	O
the	O
stereospecificity	O
for	O
glucocorticoids	O
,	O
the	O
time-course	O
of	O
steroid-receptor	O
association	O
,	O
and	O
cytoplasmic	O
to	O
nuclear	O
translocation	O
.	O

Despite	O
the	O
greater	O
number	O
of	O
glucocorticoid	B-protein
receptor	I-protein
sites	O
,	O
the	O
peanut-negative	B-cell_line
thymocyte	I-cell_line
subpopulation	I-cell_line
did	O
not	O
differ	O
from	O
the	O
peanut-positive	B-cell_line
one	O
in	O
its	O
sensitivity	O
to	O
the	O
inhibitory	O
effects	O
of	O
triamcinolone	O
acetonide	O
,	O
as	O
determined	O
by	O
measurements	O
of	O
the	O
incorporation	O
of	O
radiolabeled	O
precursors	O
of	O
protein	O
and	O
DNA	O
.	O

Moreover	O
,	O
the	O
peanut-negative	B-cell_line
subset	I-cell_line
appeared	O
more	O
resistant	O
in	O
vitro	O
to	O
the	O
steroid-induced	O
cell	O
lysis	O
as	O
compared	O
to	O
the	O
peanut-positive	B-cell_line
one	O
.	O

Thus	O
,	O
our	O
data	O
suggest	O
that	O
glucocorticoid	O
receptor	O
density	O
and	O
corticosensitivity	O
are	O
not	O
directly	O
correlated	O
and	O
that	O
the	O
number	O
of	O
glucocorticoid	B-protein
receptor	I-protein
sites	O
may	O
be	O
dependent	O
on	O
the	O
degree	O
of	O
immunologic	O
maturation	O

-DOCSTART-	O

Defective	O
binding	O
and	O
function	O
of	O
1	B-protein
,	I-protein
25-dihydroxyvitamin	I-protein
D3	I-protein
receptors	I-protein
in	O
peripheral	B-cell_type
mononuclear	I-cell_type
cells	I-cell_type
of	O
patients	O
with	O
end-organ	O
resistance	O
to	O
1	O
,	O
25-dihydroxyvitamin	O
D	O
.	O

Lectin-induced	O
DNA	O
synthesis	O
by	O
peripheral	B-cell_type
mononuclear	I-cell_type
cells	I-cell_type
from	O
17	O
normal	O
donors	O
was	O
inhibited	O
(	O
40-60	O
%	O
)	O
by	O
1	O
,	O
25-dihydroxyvitamin	O
D3	O
(	O
1	O
,	O
25	O
[	O
OH	O
]	O
2D3	O
)	O
at	O
physiological	O
concentrations	O
(	O
10	O
(	O
-10	O
)	O
-10	O
(	O
-9	O
)	O
M	O
)	O
.	O

The	O
lymphocytes	B-cell_type
acquire	O
specific	O
receptors	O
for	O
1	O
,	O
25	O
(	O
OH	O
)	O
2D3	O
upon	O
activation	O
by	O
the	O
lectins	B-protein
.	O

This	O
process	O
precedes	O
the	O
inhibitory	O
effect	O
of	O
1	O
,	O
25	O
(	O
OH	O
)	O
2D3	O
.	O

We	O
studied	O
lymphocytes	B-cell_type
from	O
six	O
patients	O
from	O
four	O
different	O
kindreds	O
with	O
the	O
syndrome	O
of	O
hereditary	O
end-organ	O
resistance	O
to	O
1	O
,	O
25	O
(	O
OH	O
)	O
2D	O
(	O
the	O
so-called	O
vitamin	O
D-dependent	O
rickets	O
type	O
II	O
)	O
.	O

In	O
five	O
patients	O
(	O
three	O
kindreds	O
)	O
peripheral	B-cell_type
blood	I-cell_type
mononuclear	I-cell_type
cells	I-cell_type
did	O
not	O
acquire	O
receptors	O
for	O
1	O
,	O
25	O
(	O
OH	O
)	O
2D3	O
upon	O
phytohemagglutinin-induced	O
activation	O
.	O

Moreover	O
,	O
in	O
contrast	O
to	O
normal	B-cell_type
lymphocytes	I-cell_type
,	O
the	O
mitogenic	O
stimulation	O
of	O
these	O
patients	O
'	O
lymphocytes	B-cell_type
by	O
phytohemagglutinin	B-protein
and	O
concanavalin	O
A	O
was	O
not	O
inhibited	O
by	O
1	O
,	O
25	O
(	O
OH	O
)	O
2D3	O
.	O

Activated	O
lymphocytes	B-cell_type
of	O
the	O
sixth	O
patient	O
from	O
a	O
fourth	O
kindred	O
exhibited	O
normal	O
binding	O
of	O
[	O
3H	O
]	O
1	O
,	O
25	O
(	O
OH	O
)	O
2D3	O
but	O
the	O
hormone	O
failed	O
to	O
inhibit	O
the	O
mitogenic	O
stimulation	O
.	O

A	O
similar	O
pattern	O
of	O
the	O
vitamin	O
D	O
effector	O
system	O
was	O
previously	O
observed	O
in	O
fibroblasts	O
cultured	O
from	O
skin	O
biopsies	O
of	O
the	O
same	O
group	O
of	O
patients	O
.	O

The	O
conclusions	O
from	O
these	O
findings	O
are	O
:	O
(	O
a	O
)	O
the	O
inhibition	O
of	O
mitogenic	O
stimulation	O
by	O
1	O
,	O
25	O
(	O
OH	O
)	O
2D3	O
is	O
mediated	O
by	O
specific	B-protein
functional	I-protein
receptors	I-protein
to	O
the	O
hormone	O
;	O
and	O
(	O
b	O
)	O
the	O
receptors	O
for	O
1	O
,	O
25	O
(	O
OH	O
)	O
2D3	O
in	O
mononuclear	B-cell_type
cells	I-cell_type
are	O
probably	O
controlled	O
genetically	O
by	O
the	O
same	O
mechanisms	O
as	O
the	O
effector	O
system	O
in	O
well-characterized	O
target	O
organs	O
of	O
the	O
hormone	O
,	O
such	O
as	O
intestine	O
and	O
kidney	O
.	O

-DOCSTART-	O

Glucocorticoid	B-protein
receptors	I-protein
of	O
mononuclear	B-cell_type
leukocytes	I-cell_type
from	O
myasthenia	O
gravis	O
patients	O
.	O

The	O
present	O
study	O
was	O
performed	O
to	O
analyse	O
glucocorticoid	B-protein
receptor	I-protein
(	O
GR	B-protein
)	O
binding	O
in	O
peripheral	B-cell_type
blood	I-cell_type
mononuclear	I-cell_type
leukocytes	I-cell_type
(	O
MNL	B-cell_type
)	O
from	O
39	O
myasthenia	O
gravis	O
(	O
MG	O
)	O
patients	O
(	O
unoperated	O
patients	O
(	O
n	O
=	O
13	O
)	O
,	O
thymectomized	O
patients	O
(	O
n	O
=	O
14	O
)	O
and	O
patients	O
receiving	O
glucocorticoids	O
:	O
thymectomized	O
(	O
n	O
=	O
11	O
)	O
and	O
unoperated	O
(	O
n	O
=	O
6	O
]	O
.	O

A	O
whole	O
cell	O
binding	O
assay	O
with	O
3	O
(	O
H	O
)	O
dexamethasone	O
was	O
used	O
.	O

GR	B-protein
mean	O
values	O
were	O
significantly	O
higher	O
in	O
the	O
MNL	B-cell_type
of	O
MG	O
patients	O
(	O
thymectomized	O
or	O
not	O
)	O
not	O
receiving	O
glucocorticoid	O
than	O
in	O
the	O
MNL	B-cell_type
of	O
healthy	O
donors	O
.	O

Affinity	O
was	O
within	O
the	O
normal	O
range	O
.	O

Sex	O
,	O
age	O
or	O
clinical	O
forms	O
of	O
illness	O
did	O
not	O
influence	O
the	O
results	O
.	O

In	O
patients	O
receiving	O
prednisone	O
(	O
Pd	O
)	O
the	O
GR	B-protein
values	O
were	O
significantly	O
lower	O
than	O
in	O
MG	O
patients	O
without	O
Pd	O
therapy	O
,	O
independent	O
of	O
Pd	O
dose	O
or	O
time	O
of	O
administration	O
.	O

No	O
differences	O
in	O
receptor	O
binding	O
between	O
normal	O
subjects	O
and	O
MG	O
patients	O
receiving	O
Pd	O
have	O
been	O
found	O
.	O

-DOCSTART-	O

Immunological	O
interference	O
of	O
high	O
dose	O
corticosteroids	O
.	O

High-dose	O
corticosteroids	O
(	O
HDC	O
)	O
will	O
influence	O
cellular	O
as	O
well	O
as	O
humoral	O
participants	O
of	O
the	O
immune	O
response	O
.	O

The	O
lymphoid	O
tissue	O
will	O
decrease	O
in	O
size	O
and	O
weight	O
after	O
prolonged	O
treatment	O
with	O
HDC	O
.	O

Lymphocyte	O
functions	O
will	O
be	O
impaired	O
.	O

Reduced	O
synthesis	O
of	O
B-	B-cell_type
as	I-cell_type
well	I-cell_type
as	I-cell_type
T-lymphocytes	I-cell_type
will	O
be	O
seen	O
.	O

The	O
inhibitory	O
effect	O
on	O
B-cell	B-cell_type
function	O
can	O
be	O
observed	O
both	O
as	O
decreased	O
serum	O
levels	O
of	O
immunoglobulins	B-protein
and	O
as	O
impaired	O
binding	O
of	O
antibodies	B-protein
and	O
complement	O
to	O
the	O
cellular	O
surface	O
.	O

Reduced	O
T-cell	B-cell_type
function	O
indicated	O
by	O
impaired	O
stimulation	O
by	O
PHA	B-protein
and	O
porkweed	B-protein
as	O
well	O
as	O
by	O
impaired	O
lymphokinin	O
effects	O
on	O
leukocyte	B-cell_type
migration	O
inhibition	O
has	O
been	O
reported	O
.	O

Reduced	O
lymphocyte	O
adherence	O
to	O
antigen	O
and	O
suppressed	O
lymphocyte	O
reaction	O
have	O
also	O
been	O
observed	O
.	O

Humoral	O
factors	O
involved	O
in	O
chemotaxis	O
,	O
opsonisation	O
,	O
phagocytosis	O
,	O
vascular	O
permeability	O
leading	O
to	O
leakage	O
of	O
fluid	O
and	O
cells	O
and	O
factors	O
involved	O
in	O
lysis	O
of	O
antigens	O
are	O
impaired	O
.	O

This	O
can	O
be	O
explained	O
partly	O
by	O
the	O
observed	O
reduced	O
complement	O
activation	O
via	O
the	O
alternative	O
as	O
well	O
as	O
the	O
classical	O
pathway	O
in	O
association	O
with	O
HDC	O
therapy	O
.	O

Acute	O
processes	O
with	O
increased	O
vascular	O
permeability	O
and	O
accumulation	O
of	O
leukocytes	B-cell_type
as	O
impairing	O
factors	O
could	O
be	O
influenced	O
beneficially	O
by	O
HDC	O
therapy	O
.	O

This	O
positive	O
effect	O
can	O
be	O
seen	O
in	O
treatment	O
of	O
septic	O
shock	O
or	O
rejection	O
of	O
a	O
transplant	O
.	O

However	O
,	O
if	O
sepsis	O
or	O
rejection	O
is	O
not	O
rapidly	O
reversed	O
,	O
complications	O
such	O
as	O
multisystem	O
organ	O
failure	O
and	O
bacteremia	O
are	O
prone	O
to	O
appear	O
.	O

-DOCSTART-	O

Identification	O
of	O
human	B-protein
leukemic	I-protein
glucocorticoid	I-protein
receptors	I-protein
using	O
affinity	O
labeling	O
and	O
anti-human	B-protein
glucocorticoid	I-protein
receptor	I-protein
antibodies	I-protein
.	O

Antisera	O
raised	O
against	O
human	B-protein
lymphoid	I-protein
glucocorticoid	I-protein
receptors	I-protein
were	O
used	O
in	O
combination	O
with	O
the	O
glucocorticoid	O
receptor	O
affinity	O
label	O
[	O
3H	O
]	O
dexamethasone	O
21-mesylate	O
[	O
(	O
3H	O
]	O
DM	O
)	O
to	O
identify	O
the	O
glucocorticoid	B-protein
receptors	I-protein
of	O
the	O
human	B-cell_line
B-lymphoblastoid	I-cell_line
cell	I-cell_line
line	I-cell_line
IM-9	B-cell_line
and	O
the	O
human	B-cell_line
T-cell	I-cell_line
leukemic	I-cell_line
cell	I-cell_line
line	I-cell_line
CEM-C7	B-cell_line
.	O

Antisera	O
were	O
obtained	O
following	O
immunization	O
of	O
New	O
Zealand	O
White	O
rabbits	O
with	O
[	B-protein
3H	I-protein
]	I-protein
triamcinolone	I-protein
acetonide	I-protein
[	I-protein
(	I-protein
3H	I-protein
]	I-protein
TA	I-protein
)	I-protein
-glucocorticoid	I-protein
receptor	I-protein
complexes	I-protein
partially	O
purified	O
by	O
two-stage	O
DNA-cellulose	O
chromatography	O
.	O

The	O
presence	O
of	O
anti-human	B-protein
glucocorticoid	I-protein
receptor	I-protein
antibodies	I-protein
was	O
verified	O
by	O
:	O
(	O
a	O
)	O
adsorption	O
of	O
[	B-protein
3H	I-protein
]	I-protein
TA-receptor-antibody	I-protein
complexes	I-protein
to	O
Protein	B-protein
A	I-protein
;	O
(	O
b	O
)	O
a	O
shift	O
to	O
higher	O
apparent	O
molecular	O
weight	O
in	O
the	O
elution	O
position	O
from	O
Sephacryl	O
S300	O
of	O
[	B-protein
3H	I-protein
]	I-protein
TA-receptor	I-protein
complexes	I-protein
incubated	O
with	O
immune	O
serum	O
;	O
and	O
(	O
c	O
)	O
the	O
ability	O
of	O
immune	O
serum	O
to	O
displace	O
[	B-protein
3H	I-protein
]	I-protein
TA-receptor	I-protein
complexes	I-protein
on	O
sucrose	O
gradients	O
.	O

These	O
antibodies	B-protein
also	O
recognized	O
rat	O
liver	O
and	O
murine	B-protein
S49	I-protein
cell	I-protein
glucocorticoid	I-protein
receptors	I-protein
.	O

Sodium	O
dodecyl	O
sulfate-polyacrylamide	O
gel	O
electrophoresis	O
of	O
[	O
3H	O
]	O
DM-labeled	O
IM-9	O
cytosol	O
identified	O
a	O
major	O
competable	O
band	O
with	O
a	O
molecular	O
weight	O
of	O
approximately	O
90	O
,	O
000	O
,	O
three	O
minor	O
competable	O
components	O
with	O
molecular	O
weights	O
of	O
approximately	O
78	O
,	O
000	O
,	O
approximately	O
51	O
,	O
000	O
,	O
and	O
approximately	O
38	O
,	O
500	O
,	O
and	O
at	O
least	O
21	O
other	O
noncompetable	O
components	O
.	O

Following	O
immunoprecipitation	O
of	O
[	O
3H	O
]	O
DM-labeled	O
cytosol	O
with	O
immune	O
serum	O
,	O
only	O
the	O
Mr	O
90	O
,	O
000	O
and	O
78	O
,	O
000	O
components	O
were	O
seen	O
.	O

Sodium	O
dodecyl	O
sulfate-polyacrylamide	O
gel	O
electrophoresis	O
of	O
[	O
3H	O
]	O
DM-labeled	O
CEM-C7	O
cytosol	O
revealed	O
a	O
larger	O
number	O
of	O
[	O
3H	O
]	O
DM-labeled	O
components	O
.	O

However	O
,	O
after	O
immunoprecipitation	O
of	O
[	O
3H	O
]	O
DM-labeled	O
CEM-C7	O
cytosol	O
,	O
a	O
predominant	O
competable	O
component	O
with	O
a	O
molecular	O
weight	O
of	O
90	O
,	O
000	O
was	O
easily	O
identified	O
.	O

This	O
component	O
was	O
markedly	O
diminished	O
when	O
cytosols	O
from	O
the	O
glucocorticoid	B-cell_line
receptor-deficient	I-cell_line
cell	I-cell_line
line	I-cell_line
ICR-27	B-cell_line
were	O
used	O
.	O

Thus	O
,	O
the	O
combination	O
of	O
affinity	O
labeling	O
and	O
anti-human	B-protein
glucocorticoid	I-protein
receptor	I-protein
antibodies	I-protein
is	O
capable	O
of	O
providing	O
direct	O
physical	O
identification	O
of	O
human	B-protein
lymphoid	I-protein
glucocorticoid	I-protein
receptors	I-protein
.	O

-DOCSTART-	O

Effect	O
of	O
thymosin	O
on	O
glucocorticoid	B-protein
receptor	I-protein
activity	O
and	O
glucocorticoid	O
sensitivity	O
of	O
human	B-cell_type
thymocytes	I-cell_type
.	O

Incubation	O
with	O
thymosin	O
fraction	O
5	O
,	O
(	O
TMS	O
F5	O
at	O
300	O
micrograms/ml	O
)	O
a	O
partially	O
purified	O
thymic	O
factor	O
,	O
reduced	O
the	O
steroid	O
binding	O
activity	O
of	O
human	B-cell_type
infant	I-cell_type
thymocytes	I-cell_type
from	O
9.6	O
+/-	O
2.1	O
fmole/ml	O
to	O
5.0	O
+/-	O
2.0	O
fmole/ml	O
.	O

The	O
glucocorticoid	B-protein
receptor	I-protein
activity	O
in	O
normal	O
infant	O
thymocytes	O
was	O
found	O
to	O
be	O
2	O
,	O
146	O
+/-	O
726	O
(	O
s.d.	O
)	O
sites	O
per	O
cell	O
with	O
dissociation	O
constant	O
of	O
1.4	O
+/-	O
0.6	O
X	O
10	O
(	O
-8	O
)	O
M	O
.	O

TMS	O
F5	O
also	O
increased	O
the	O
resistance	O
of	O
human	O
thymocytes	O
to	O
the	O
cytolytic	O
effect	O
of	O
dexamethasone	O
(	O
2.5	O
X	O
10	O
(	O
-8	O
)	O
M	O
)	O
to	O
168.6	O
+/-	O
30.2	O
%	O
of	O
control	O
(	O
P	O
less	O
than	O
0.01	O
)	O
.	O

In	O
animals	O
,	O
medullary	B-cell_type
and	I-cell_type
peripheral	I-cell_type
blood	I-cell_type
T	I-cell_type
cells	I-cell_type
are	O
more	O
resistant	O
to	O
glucocorticoids	O
than	O
immature	B-cell_type
thymic	I-cell_type
T	I-cell_type
cells	I-cell_type
.	O

The	O
results	O
show	O
that	O
thymosin	O
can	O
induce	O
changes	O
consistent	O
with	O
differentiation	O
in	O
human	B-cell_type
thymocytes	I-cell_type
.	O

These	O
in	O
vitro	O
results	O
are	O
consistent	O
with	O
a	O
physiological	O
role	O
of	O
thymosin	O
in	O
intrathymic	O
T	O
cell	O
maturation	O
in	O
man	O
.	O

Incubation	O
of	O
a	O
human	B-cell_line
malignant	I-cell_line
thymus	I-cell_line
derived	I-cell_line
T	I-cell_line
cell	I-cell_line
line	I-cell_line
(	O
MOLT	B-cell_line
3	I-cell_line
)	O
with	O
TMS	O
F5	O
also	O
resulted	O
in	O
a	O
significant	O
reduction	O
of	O
the	O
number	O
of	O
steroid	O
binding	O
sites	O
to	O
44.2	O
+/-	O
15.3	O
%	O
of	O
control	O
(	O
P	O
less	O
than	O
0.05	O
)	O
,	O
but	O
TMS	O
F5	O
did	O
not	O
significantly	O
reduce	O
the	O
glucocorticoid	O
sensitivity	O
of	O
MOLT	B-cell_line
3	I-cell_line
cells	I-cell_line
.	O

-DOCSTART-	O

Specific	B-protein
high-affinity	I-protein
receptors	I-protein
for	O
1	O
,	O
25-dihydroxyvitamin	O
D3	O
in	O
human	B-cell_type
peripheral	I-cell_type
blood	I-cell_type
mononuclear	I-cell_type
cells	I-cell_type
:	O
presence	O
in	O
monocytes	B-cell_type
and	O
induction	O
in	O
T	B-cell_type
lymphocytes	I-cell_type
following	O
activation	O
.	O

Human	B-cell_type
peripheral	I-cell_type
blood	I-cell_type
monocytes	I-cell_type
have	O
high	O
affinity	O
binding	O
sites	O
for	O
1	O
,	O
25-	O
(	O
OH	O
)	O
2D3	O
(	O
Kd	O
0.14	O
nM	O
,	O
sedimentation	O
coefficient	O
3.7S	O
)	O
.	O

Resting	B-cell_type
human	I-cell_type
peripheral	I-cell_type
blood	I-cell_type
T	I-cell_type
lymphocytes	I-cell_type
,	O
however	O
,	O
do	O
not	O
have	O
a	O
demonstrable	O
1	B-protein
,	I-protein
25-	I-protein
(	I-protein
OH	I-protein
)	I-protein
2D3	I-protein
receptor	I-protein
.	O

After	O
activation	O
with	O
phytohemagglutinin	B-protein
the	O
T	B-cell_type
cells	I-cell_type
exhibit	O
the	O
receptor	O
within	O
24	O
h	O
,	O
and	O
this	O
expression	O
is	O
blocked	O
by	O
cycloheximide	O
.	O

The	O
receptor	O
in	O
activated	B-cell_type
T	I-cell_type
lymphocytes	I-cell_type
has	O
a	O
sedimentation	O
coefficient	O
of	O
3.7S	O
and	O
a	O
high	O
affinity	O
(	O
Kd	O
0.10	O
nM	O
)	O
for	O
the	O
ligand	O
.	O

-DOCSTART-	O

Effects	O
of	O
chronic	O
glucocorticoid	O
excess	O
in	O
man	O
on	O
insulin	O
binding	O
to	O
circulating	B-cell_type
cells	I-cell_type
:	O
differences	O
between	O
endogenous	O
and	O
exogenous	O
hypercorticism	O
.	O

We	O
measured	O
[	O
125I	O
]	O
insulin	O
binding	O
to	O
circulating	B-cell_type
monocytes	I-cell_type
or	I-cell_type
erythrocytes	I-cell_type
from	O
16	O
patients	O
with	O
chronic	O
glucocorticoid	O
excess	O
,	O
9	O
chronically	O
treated	O
with	O
prednisone	O
and	O
7	O
with	O
adrenocortical	O
hyperfunction	O
.	O

With	O
monocytes	B-cell_type
,	O
[	O
125I	O
]	O
insulin	O
binding	O
was	O
iincreased	O
in	O
all	O
patients	O
.	O

Analysis	O
of	O
binding	O
data	O
indicated	O
that	O
increased	O
binding	O
in	O
patients	O
treated	O
with	O
prednisone	O
was	O
due	O
to	O
an	O
ncrease	O
in	O
receptor	O
concentration	O
,	O
whereas	O
in	O
patients	O
with	O
adrenocortical	O
hyperfunction	O
,	O
it	O
was	O
due	O
to	O
an	O
increase	O
in	O
receptor	O
affinity	O
.	O

With	O
erythrocytes	O
from	O
patients	O
with	O
adrenocortical	O
hyperfunction	O
there	O
was	O
an	O
increase	O
in	O
receptor	O
affinity	O
and	O
a	O
decrease	O
in	O
receptor	O
concentration	O
,	O
so	O
that	O
the	O
binding	O
of	O
[	O
125I	O
]	O
insulin	O
was	O
normal	O
.	O

The	O
disparity	O
of	O
results	O
between	O
endogenous	O
and	O
exogenous	O
hypercorticism	O
,	O
between	O
the	O
two	O
cell	O
types	O
,	O
and	O
between	O
the	O
present	O
studies	O
and	O
previous	O
studies	O
suggest	O
that	O
the	O
effects	O
of	O
glucocorticoid	O
excess	O
on	O
the	O
insulin	B-protein
receptor	I-protein
are	O
extremely	O
complex	O
and	O
wide-ranging	O
and	O
that	O
in	O
this	O
condition	O
,	O
extrapolations	O
in	O
humans	O
from	O
data	O
with	O
circulating	B-cell_type
cells	I-cell_type
to	O
liver	O
and	O
muscle	O
may	O
not	O
be	O
appropriate	O
.	O

-DOCSTART-	O

Reduced	O
level	O
of	O
cellular	B-protein
glucocorticoid	I-protein
receptors	I-protein
in	O
patients	O
with	O
anorexia	O
nervosa	O
.	O

Specific	O
glucocorticoid	B-protein
receptors	I-protein
were	O
measured	O
in	O
circulating	O
mononuclear	B-cell_type
leukocytes	I-cell_type
from	O
12	O
patients	O
with	O
anorexia	O
nervosa	O
and	O
21	O
healthy	O
control	O
subjects	O
.	O

Cells	O
from	O
patients	O
were	O
found	O
to	O
contain	O
a	O
significantly	O
(	O
p	O
less	O
than	O
0.01	O
)	O
lower	O
level	O
of	O
glucocorticoid	B-protein
receptor	I-protein
(	O
3830	O
+/-	O
210	O
sites/cell	O
,	O
mean	O
+/-	O
SE	O
)	O
than	O
those	O
from	O
controls	O
(	O
4930	O
+/-	O
250	O
sites/cell	O
)	O
.	O

A	O
partial	O
glucocorticoid	B-protein
receptor	I-protein
defect	O
may	O
well	O
explain	O
the	O
abnormal	O
cortisol	O
metabolism	O
and	O
glucocorticoid	O
resistance	O
commonly	O
found	O
in	O
patients	O
with	O
anorexia	O
nervosa	O
.	O

-DOCSTART-	O

Regulation	O
of	O
the	O
glucocorticoid	B-protein
receptor	I-protein
in	O
human	B-cell_type
lymphocytes	I-cell_type
.	O

The	O
presence	O
of	O
a	O
glucocorticoid	B-protein
receptor	I-protein
in	O
human	B-cell_type
lymphocytes	I-cell_type
is	O
well	O
established	O
,	O
but	O
factors	O
affecting	O
its	O
regulation	O
have	O
not	O
been	O
described	O
.	O

Using	O
a	O
competitive	O
binding	O
whole	O
cell	O
assay	O
,	O
we	O
have	O
examined	O
the	O
binding	O
of	O
[	O
3H	O
]	O
-dexamethasone	O
at	O
24	O
and	O
37	O
degrees	O
C	O
in	O
untreated	O
normal	O
subjects	O
and	O
in	O
healthy	O
subjects	O
taking	O
various	O
glucocorticoid	O
preparations	O
.	O

At	O
24	O
degrees	O
C	O
normal	B-cell_type
human	I-cell_type
lymphocytes	I-cell_type
had	O
6000	O
binding	O
sites/cell	O
and	O
a	O
dissociation	O
constant	O
of	O
4	O
x	O
10	O
(	O
-9	O
)	O
M	O
.	O

The	O
administration	O
of	O
1	O
mg	O
of	O
dexamethasone	O
,	O
5	O
mg	O
of	O
prednisone	O
,	O
and	O
37.5	O
mg	O
of	O
cortisone	O
acetate	O
resulted	O
in	O
a	O
30	O
%	O
decrease	O
in	O
binding	O
sites	O
after	O
1	O
week	O
with	O
no	O
change	O
in	O
binding	O
affinity	O
.	O

No	O
changes	O
in	O
the	O
number	O
of	O
binding	O
sites	O
was	O
noted	O
before	O
1	O
week	O
and	O
the	O
diminished	O
number	O
persisted	O
for	O
1	O
week	O
after	O
discontinuation	O
of	O
glucocorticoid	O
treatment	O
.	O

Lymphocytes	B-cell_type
from	O
hospitalized	O
patients	O
taking	O
40-60	O
mg	O
of	O
dexamethasone	O
daily	O
demonstrated	O
the	O
same	O
change	O
in	O
number	O
of	O
binding	O
sites	O
that	O
was	O
seen	O
in	O
normal	O
subjects	O
taking	O
1	O
mg	O
of	O
dexamethasone	O
.	O

When	O
binding	O
assays	O
were	O
carried	O
out	O
at	O
physiologic	O
temperature	O
there	O
was	O
the	O
same	O
decrease	O
in	O
number	O
of	O
binding	O
sites	O
after	O
dexamethasone	O
administration	O
,	O
and	O
in	O
addition	O
,	O
there	O
was	O
a	O
two-fold	O
increase	O
in	O
binding	O
affinity	O
.	O

Glucocorticoid	O
administration	O
results	O
in	O
a	O
time-dependent	O
decrease	O
in	O
the	O
number	O
of	O
lymphocyte	O
glucocorticoid	O
binding	O
sites	O
that	O
is	O
independent	O
of	O
the	O
type	O
of	O
glucocorticoid	O
administered	O
.	O

This	O
is	O
the	O
first	O
in	O
vivo	O
demonstration	O
that	O
glucocorticoids	O
modulate	O
their	O
own	O
receptors	O
in	O
man	O
.	O

-DOCSTART-	O

Immunoglobulin	O
localization	O
in	O
benign	O
and	O
malignant	O
lesions	O
of	O
the	O
human	O
mammary	O
gland	O
.	O

Using	O
direct	O
imunofluorescence	O
,	O
lesions	O
from	O
266	O
human	O
breast	O
specimens	O
were	O
studied	O
for	O
the	O
presence	O
of	O
IgA	O
,	O
IgM	O
,	O
or	O
IgG	O
localization	O
.	O

The	O
lesions	O
included	O
benign	O
elements	O
from	O
66	O
subcutaneous	O
mastectomy	O
specimens	O
in	O
which	O
the	O
absence	O
of	O
simultaneous	O
breast	O
malignancy	O
was	O
documented	O
,	O
primary	O
breast	O
carcinomas	O
from	O
153	O
mastectomy	O
specimens	O
,	O
and	O
47	O
biopsies	O
containing	O
metastatic	O
breast	O
cancer	O
.	O

A	O
statistically	O
significant	O
association	O
of	O
IgA	B-protein
and	O
IgM	B-protein
with	O
benign	O
lesions	O
was	O
contrasted	O
to	O
the	O
association	O
of	O
IgG	B-protein
with	O
malignant	O
lesions	O
.	O

In	O
both	O
primary	O
and	O
metastatic	O
lesions	O
,	O
IgG	B-protein
localization	O
was	O
associated	O
with	O
estrogen-receptor-poor	O
primary	O
cancers	O
as	O
compared	O
with	O
estrogen-receptor-rich	O
primary	O
cancers	O
.	O

Among	O
primary	O
breast	O
cancer	O
patients	O
,	O
IgG	B-protein
localization	O
in	O
the	O
tumor	O
correlated	O
with	O
relative	O
lymphopenia	O
.	O

A	O
shorter	O
disease-free	O
interval	O
was	O
noted	O
in	O
association	O
with	O
IgG	B-protein
localization	O
among	O
the	O
metastatic	O
breast	O
lesions	O
.	O

No	O
statistically	O
significant	O
association	O
between	O
stage	O
of	O
disease	O
and	O
immunoglobulin	O
presence	O
was	O
demonstrable	O
.	O

Moderate-to-severe	O
intraductal	O
epithelial	O
hyperplasias	O
were	O
more	O
often	O
associated	O
with	O
immunoglobulin	B-protein
G	I-protein
localization	O
that	O
were	O
other	O
benign	O
lesions	O

-DOCSTART-	O

Correlation	O
of	O
steroid	B-protein
receptors	I-protein
with	O
histologic	O
differentiation	O
in	O
mammary	O
carcinoma	O
.	O

A	O
Singapore	O
experience	O
.	O

Cancer	O
of	O
the	O
breast	O
is	O
the	O
most	O
common	O
tumor	O
in	O
females	O
in	O
Singapore	O
,	O
with	O
the	O
rate	O
of	O
20.7	O
per	O
100	O
,	O
000	O
per	O
year	O
(	O
1977	O
estimate	O
)	O
,	O
which	O
is	O
predicted	O
to	O
increase	O
to	O
29.8	O
per	O
100	O
,	O
000	O
women	O
per	O
year	O
by	O
1995	O
.	O

A	O
detailed	O
histopathologic	O
review	O
of	O
50	O
primary	O
breast	O
cancer	O
tumors	O
analyzed	O
for	O
estrogen	B-protein
receptor	I-protein
(	O
ER	B-protein
)	O
level	O
was	O
carried	O
out	O
and	O
a	O
variety	O
of	O
morphologic	O
features	O
correlated	O
with	O
ER	B-protein
results	O
to	O
identify	O
any	O
factors	O
that	O
will	O
improve	O
the	O
management	O
and	O
prognosis	O
for	O
breast	O
cancer	O
.	O

Cytosol	O
was	O
incubated	O
with	O
3H-estradiol	O
in	O
the	O
presence	O
and	O
absence	O
of	O
cold	O
diethylstilbestrol	O
,	O
and	O
bound	O
and	O
free	O
hormone	O
were	O
separated	O
by	O
Dextran-coated	O
charcoal	O
method	O
.	O

Tumors	O
binding	O
more	O
than	O
5	O
fmol/mg	O
cytosol	B-protein
protein	I-protein
were	O
classified	O
as	O
ER	B-protein
-positive	O
.	O

Progesterone	B-protein
receptor	I-protein
(	O
PR	B-protein
)	O
level	O
was	O
analyzed	O
in	O
some	O
specimens	O
with	O
the	O
use	O
of	O
a	O
similar	O
method	O
.	O

Most	O
of	O
the	O
patients	O
were	O
Chinese	O
(	O
90	O
%	O
)	O
.	O

Three	O
patients	O
were	O
Malays	O
,	O
one	O
was	O
Indian	O
,	O
and	O
one	O
was	O
European	O
in	O
this	O
series	O
.	O

Results	O
indicated	O
that	O
there	O
was	O
strong	O
correlation	O
between	O
ER	B-protein
level	O
,	O
age	O
,	O
and	O
histologic	O
grade	O
of	O
the	O
tumors	O
.	O

No	O
correlation	O
existed	O
between	O
absence	O
or	O
presence	O
of	O
lymph	O
node	O
metastases	O
and	O
ER	B-protein
.	O

Although	O
there	O
was	O
a	O
trend	O
for	O
ER	B-protein
-positive	O
tumors	O
to	O
have	O
a	O
low-grade	O
lymphocytic	O
infiltration	O
,	O
the	O
difference	O
was	O
not	O
statistically	O
significant	O
.	O

-DOCSTART-	O

Mononuclear	B-cell_type
cells	I-cell_type
infiltrating	O
human	O
mammary	O
carcinomas	O
:	O
immunohistochemical	O
analysis	O
with	O
monoclonal	B-protein
antibodies	I-protein
.	O

Breast	O
carcinomas	O
were	O
examined	O
by	O
the	O
immunoperoxidase	O
technique	O
using	O
antisera	O
specific	O
for	O
lymphocyte	B-cell_type
subsets	I-cell_type
,	O
monocytes	B-cell_type
,	O
NK	B-cell_type
cells	I-cell_type
and	O
major	B-protein
histocompatibility	I-protein
antigens	I-protein
(	O
HLA-A	B-protein
,	O
-B	B-protein
,	O
-C	O
;	O
Ia-like	B-protein
)	O
.	O

Sixty-four	O
per	O
cent	O
of	O
the	O
patients	O
had	O
a	O
moderate	O
or	O
strong	O
mononuclear	O
cell	O
infiltration	O
,	O
77	O
%	O
of	O
the	O
patients	O
without	O
mononuclear	B-cell_type
cell	I-cell_type
infiltration	O
had	O
receptors	O
for	O
estrogens	O
as	O
compared	O
to	O
51	O
%	O
of	O
the	O
patients	O
with	O
infiltration	O
.	O

The	O
majority	O
of	O
the	O
infiltrating	B-cell_type
mononuclear	I-cell_type
cells	I-cell_type
were	O
T	B-cell_type
cells	I-cell_type
;	O
generally	O
the	O
OKT8	B-cell_line
cells	I-cell_line
were	O
predominant	O
.	O

The	O
Leu	O
3A/OKT8	O
cell	O
ratio	O
was	O
not	O
related	O
to	O
histological	O
type	O
,	O
tumor	O
size	O
,	O
age	O
of	O
the	O
patient	O
or	O
presence	O
of	O
metastases	O
.	O

Some	O
of	O
the	O
T	O
cells	O
had	O
the	O
Ia	B-protein
antigen	I-protein
and	O
were	O
thus	O
probably	O
activated	O
.	O

The	O
B	B-cell_type
cells	I-cell_type
were	O
either	O
absent	O
or	O
less	O
numerous	O
than	O
the	O
T	O
cells	O
.	O

There	O
was	O
no	O
relation	O
between	O
their	O
distribution	O
and	O
the	O
various	O
parameters	O
studied	O
.	O

A	O
few	O
monocytes	B-cell_type
were	O
heterogeneous	O
according	O
to	O
their	O
markers	O
(	O
OKM	B-protein
I	I-protein
and	O
acid	O
phosphatase	O
)	O
.	O

In	O
6	O
cases	O
only	O
there	O
was	O
a	O
strong	O
infiltration	O
of	O
mononuclear	B-cell_type
cells	I-cell_type
positive	O
for	O
acid	B-protein
phosphatase	I-protein
.	O

The	O
number	O
of	O
the	O
natural	B-cell_type
killer	I-cell_type
cells	I-cell_type
was	O
also	O
low	O
.	O

Only	O
a	O
few	O
mononuclear	O
infiltrating	O
cells	O
had	O
receptors	O
for	O
transferrin	O
.	O

There	O
was	O
a	O
positive	O
correlation	O
between	O
the	O
inflammatory	O
infiltration	O
and	O
the	O
presence	O
of	O
HLA	B-protein
class-I	I-protein
antigens	I-protein
on	O
tumor	B-cell_type
cell	I-cell_type
s	O
.	O

Some	O
of	O
the	O
antisera	O
specific	O
for	O
lymphocyte	B-cell_type
subsets	I-cell_type
also	O
stained	O
the	O
breast	B-cell_type
carcinoma	I-cell_type
cells	I-cell_type
.	O

The	O
great	O
variations	O
in	O
the	O
subsets	O
of	O
mononuclear	B-cell_type
cells	I-cell_type
in	O
breast	O
carcinomas	O
may	O
correspond	O
to	O
various	O
systems	O
of	O
defense	O
against	O
neoplasm	O
.	O

-DOCSTART-	O

A	O
case	O
of	O
male	O
pseudohermaphroditism	O
with	O
normal	O
androgen	B-protein
receptor	I-protein
binding	O
and	O
47	O
,	O
XYY	O
karyotype	O
.	O

A	O
case	O
of	O
male	O
pseudohermaphroditism	O
with	O
47	O
,	O
XYY	O
karyotype	O
in	O
blood	O
and	O
cutaneous	B-cell_type
fibroblasts	I-cell_type
is	O
described	O
.	O

The	O
plasma	O
testosterone	O
response	O
to	O
HCG	O
stimulation	O
was	O
slightly	O
below	O
the	O
normal	O
range	O
on	O
two	O
occasions	O
suggesting	O
a	O
deficit	O
of	O
gonadal	O
function	O
.	O

A	O
study	O
of	O
the	O
receptors	O
for	O
dihydrotestosterone	O
in	O
fibroblasts	B-cell_type
of	O
genital	O
and	O
nongenital	O
skin	O
showed	O
a	O
normal	O
concentration	O
of	O
receptors	O
in	O
genital	O
skin	O
;	O
5-alpha-reductase	B-protein
activity	O
in	O
fibroblasts	B-cell_type
of	O
the	O
genital	O
skin	O
was	O
low	O
,	O
but	O
the	O
plasma	O
relationship	O
testosterone/dihydrotestosterone	O
under	O
HCG	O
stimulation	O
was	O
normal	O
.	O

The	O
diagnostic	O
possibility	O
of	O
a	O
complete	O
testicular	O
feminization	O
syndrome	O
with	O
normal	O
receptors	O
for	O
dihydrotestosterone	O
is	O
commented	O
on	O
.	O

-DOCSTART-	O

1	O
,	O
25-Dihydroxyvitamin	O
D3	O
inhibits	O
antigen-induced	O
T	B-cell_type
cell	I-cell_type
activation	O
.	O

The	O
proliferative	O
response	O
of	O
murine	B-cell_type
spleen	I-cell_type
and	I-cell_type
thymus	I-cell_type
cells	I-cell_type
to	O
antigen	B-protein
but	O
not	O
to	O
lectin	B-protein
was	O
inhibited	O
by	O
the	O
active	O
metabolite	O
of	O
vitamin	O
D3	O
,	O
1	O
,	O
25-	O
(	O
OH	O
)	O
2D3	O
.	O

To	O
directly	O
examine	O
the	O
effect	O
of	O
1	O
,	O
25-	O
(	O
OH	O
)	O
2D3	O
on	O
T	B-cell_type
cell	I-cell_type
activation	O
in	O
the	O
absence	O
of	O
other	O
complicating	O
interactions	O
,	O
we	O
utilized	O
a	O
panel	O
of	O
cloned	O
Ia-restricted	B-cell_line
T	I-cell_line
cell	I-cell_line
hybridomas	I-cell_line
that	O
secrete	O
IL	B-protein
2	I-protein
on	O
activation	O
by	O
cloned	O
Ia-bearing	B-cell_type
stimulator	I-cell_type
cells	I-cell_type
(	O
TA3	B-cell_type
)	O
or	O
when	O
stimulated	O
by	O
mitogen	O
.	O

Physiologic	O
concentrations	O
of	O
1	O
,	O
25-	O
(	O
OH	O
)	O
2D3	O
(	O
0.01	O
to	O
0.1	O
nm	O
)	O
inhibited	O
the	O
antigen-induced	O
secretion	O
of	O
IL	O
2	O
by	O
several	O
of	O
these	O
T	B-cell_line
cell	I-cell_line
hybridomas	I-cell_line
.	O

This	O
inhibition	O
was	O
dependent	O
on	O
the	O
concentration	O
of	O
the	O
free	O
hormone	O
and	O
could	O
be	O
overcome	O
by	O
increasing	O
the	O
number	O
of	O
Ia-bearing	B-cell_line
stimulator	I-cell_line
cells	I-cell_line
used	O
.	O

Pretreatment	O
of	O
the	O
T	B-cell_line
hybridoma	I-cell_line
but	O
not	O
the	O
TA3	B-cell_line
stimulator	I-cell_line
cell	I-cell_line
with	O
1	O
,	O
25-	O
(	O
OH	O
)	O
2D3	O
resulted	O
in	O
inhibition	O
of	O
activation	O
.	O

These	O
results	O
are	O
consistent	O
with	O
the	O
finding	O
that	O
specific	O
1	B-protein
,	I-protein
25-	I-protein
(	I-protein
OH	I-protein
)	I-protein
2D3	I-protein
receptors	I-protein
are	O
present	O
on	O
the	O
T	O
cell	O
hybridomas	O
but	O
are	O
lacking	O
in	O
TA3	B-cell_line
cells	I-cell_line
.	O

1	O
,	O
25-	O
(	O
OH	O
)	O
2D3	O
failed	O
,	O
however	O
,	O
to	O
inhibit	O
the	O
activation	O
of	O
the	O
T	B-cell_line
cell	I-cell_line
hybridomas	I-cell_line
by	O
lectin	B-protein
or	O
by	O
an	O
anti-Thy-1	B-protein
antibody	I-protein
.	O

These	O
findings	O
suggest	O
that	O
1	O
,	O
25-	O
(	O
OH	O
)	O
2D3	O
may	O
be	O
interfering	O
with	O
early	O
events	O
of	O
antigen-induced	O
T	B-cell_type
cell	I-cell_type
activation	O
,	O
perhaps	O
by	O
hindering	O
T	B-cell_type
cell	I-cell_type
recognition	O
of	O
the	O
relevant	O
antigen	O
on	O
stimulator	O
cell	O
surfaces	O
.	O

This	O
system	O
should	O
prove	O
useful	O
in	O
studying	O
the	O
molecular	O
mechanisms	O
by	O
which	O
1	O
,	O
25-	O
(	O
OH	O
)	O
2D3	O
acts	O
to	O
inhibit	O
T	O
cell	O
activation	O
and	O
subsequent	O
IL	B-protein
2	I-protein
production	O
.	O

-DOCSTART-	O

Glucocorticoid	B-protein
receptors	I-protein
and	O
steroid	O
sensitivity	O
in	O
normal	O
and	O
neoplastic	O
human	O
lymphoid	O
tissues	O
:	O
a	O
review	O
.	O

The	O
determination	O
of	O
estrogen	B-protein
and	I-protein
progesterone	I-protein
receptors	I-protein
in	O
breast	O
cancer	O
has	O
been	O
shown	O
to	O
be	O
useful	O
in	O
predicting	O
the	O
response	O
to	O
endocrine	O
therapy	O
.	O

Given	O
their	O
well-known	O
inhibitory	O
effects	O
on	O
lymphoid	O
tissue	O
,	O
glucocorticoids	O
have	O
been	O
used	O
widely	O
in	O
the	O
treatment	O
of	O
leukemia	O
.	O

Given	O
these	O
facts	O
,	O
over	O
the	O
last	O
10	O
years	O
,	O
several	O
investigators	O
have	O
measured	O
the	O
number	O
of	O
glucocorticoid	B-protein
receptors	I-protein
in	O
normal	O
and	O
neoplastic	O
lymphoid	O
tissue	O
to	O
see	O
whether	O
their	O
number	O
correlated	O
with	O
glucocorticoid	O
responsiveness	O
in	O
vitro	O
or	O
in	O
vivo	O
.	O

No	O
clear	O
correlation	O
could	O
be	O
established	O
between	O
the	O
level	O
of	O
glucocorticoid	B-protein
receptor	I-protein
and	O
the	O
in	O
vitro	O
action	O
of	O
steroids	O
in	O
normal	O
and	O
neoplastic	O
lymphoid	O
tissue	O
.	O

In	O
contrast	O
,	O
attempts	O
to	O
correlate	O
glucocorticoid	B-protein
receptor	I-protein
levels	O
in	O
acute	O
lymphocytic	O
leukemia	O
to	O
in	O
vivo	O
steroid	O
responsiveness	O
and	O
immunological	O
type	O
using	O
the	O
whole-cell-binding	O
assay	O
for	O
receptor	O
determination	O
and	O
selecting	O
the	O
patients	O
according	O
to	O
age	O
and	O
immunological	O
criteria	O
have	O
been	O
more	O
successful	O
.	O

-DOCSTART-	O

[	O
Glucocorticoid	B-protein
receptors	I-protein
in	O
normal	B-cell_type
human	I-cell_type
lymphocytes	I-cell_type
]	O

Glucocorticoid	B-cell_type
(	I-cell_type
GC	I-cell_type
)	I-cell_type
receptors	I-cell_type
were	O
studied	O
in	O
intact	O
lymphocytes	B-cell_type
from	O
11	O
donors	O
.	O

GC	O
binding	O
parameters	O
were	O
found	O
to	O
be	O
highly	O
reproducible	O
in	O
repeated	O
experiments	O
with	O
lymphocytes	B-cell_type
.	O

It	O
was	O
shown	O
that	O
GC	B-protein
receptors	I-protein
in	O
donors	O
'	O
lymphocytes	B-cell_type
could	O
be	O
distributed	O
into	O
two	O
different	O
classes	O
similarly	O
to	O
the	O
pattern	O
seen	O
in	O
skin	B-cell_type
fibroblasts	I-cell_type
.	O

Human	B-cell_type
lymphocytes	I-cell_type
are	O
an	O
adequate	O
object	O
for	O
studying	O
genetically	O
determined	O
variability	O
of	O
GC	B-protein
receptors	I-protein
and	O
its	O
clinical	O
importance	O
.	O

-DOCSTART-	O

Specific	O
estrogen	B-protein
binding	I-protein
sites	I-protein
in	O
human	B-cell_type
lymphoid	I-cell_type
cells	I-cell_type
and	O
thymic	B-cell_type
cells	I-cell_type
.	O

The	O
binding	O
of	O
estrogen	O
in	O
preparations	O
of	O
human	B-cell_type
peripheral	I-cell_type
blood	I-cell_type
mononuclear	I-cell_type
cells	I-cell_type
,	I-cell_type
as	O
well	O
as	O
by	O
splenic	B-cell_type
and	I-cell_type
thymic	I-cell_type
cells	I-cell_type
is	O
demonstrated	O
by	O
three	O
different	O
approaches	O
(	O
Dextran-coated	O
charcoal	O
method	O
,	O
whole	O
cell	O
assay	O
,	O
and	O
gel	O
filtration	O
on	O
a	O
sepharose	O
4B	O
column	O
)	O
.	O

Scatchard	O
's	O
analysis	O
of	O
[	O
3H	O
]	O
-moxestrol	O
(	O
R2858	O
)	O
and	O
[	O
3H	O
]	O
-estradiol	O
binding	O
proves	O
the	O
existence	O
of	O
a	O
single	O
class	O
of	O
receptor	B-protein
sites	I-protein
having	O
a	O
dissociation	O
constant	O
of	O
0.18-2.4	O
X	O
10	O
(	O
-9	O
)	O
M	O
.	O

Physicochemical	O
properties	O
of	O
the	O
binder	O
,	O
including	O
binding	O
capacity	O
and	O
steroid	O
specificity	O
,	O
are	O
quite	O
similar	O
to	O
those	O
reported	O
for	O
the	O
thymus	O
of	O
small	O
mammalian	O
species	O
or	O
human	O
thymoma	O
.	O

-DOCSTART-	O

Administration	O
of	O
fibroblast	B-protein
interferon	I-protein
to	O
patients	O
with	O
advanced	O
breast	O
cancer	O
:	O
possible	O
effects	O
on	O
skin	O
metastasis	O
and	O
on	O
hormone	B-protein
receptors	I-protein
.	O

Eleven	O
patients	O
with	O
metastasized	O
breast	O
cancer	O
received	O
8	O
intramuscular	O
injections	O
of	O
6	O
x	O
10	O
(	O
6	O
)	O
units	O
of	O
human	B-protein
fibroblast	I-protein
interferon	I-protein
over	O
a	O
period	O
of	O
40	O
days	O
.	O

The	O
injections	O
did	O
not	O
cause	O
local	O
irritation	O
or	O
inflammation	O
.	O

Fever	O
occurred	O
in	O
only	O
1	O
of	O
the	O
11	O
patients	O
.	O

Although	O
several	O
types	O
of	O
metastases	O
were	O
monitored	O
,	O
only	O
skin	O
nodules	O
consistently	O
(	O
10	O
out	O
of	O
11	O
patients	O
)	O
exhibited	O
changes	O
that	O
were	O
suggestive	O
of	O
a	O
therapeutic	O
effect	O
of	O
the	O
treatment	O
regimen	O
:	O
either	O
a	O
simple	O
decrease	O
in	O
size	O
of	O
some	O
nodules	O
or	O
central	O
necrosis	O
accompanied	O
by	O
an	O
inflammatory	O
reaction	O
.	O

NK-activity	O
of	O
peripheral	B-cell_type
blood	I-cell_type
leukocytes	I-cell_type
was	O
significantly	O
increased	O
after	O
administration	O
of	O
the	O
first	O
dose	O
;	O
the	O
effect	O
of	O
subsequent	O
injections	O
was	O
less	O
clear	O
.	O

Receptors	O
for	O
estrogens	O
and	O
progestogens	O
were	O
increased	O
in	O
the	O
tumor	O
biopsies	O
of	O
2	O
out	O
of	O
2	O
and	O
5	O
out	O
of	O
6	O
patients	O
tested	O
respectively	O
.	O

-DOCSTART-	O

Decreased	O
glucocorticoid	B-protein
receptor	I-protein
binding	O
in	O
adrenal	O
insufficiency	O
.	O

To	O
examine	O
the	O
effect	O
of	O
glucocorticoid	O
deficiency	O
on	O
the	O
glucocorticoid	B-protein
receptor	I-protein
,	O
we	O
examine	O
the	O
binding	O
of	O
[	O
3H	O
]	O
dexamethasone	O
to	O
lymphocytes	B-cell_type
in	O
normal	O
subjects	O
and	O
patients	O
with	O
adrenal	O
insufficiency	O
before	O
and	O
after	O
glucocorticoid	O
replacement	O
therapy	O
.	O

Using	O
a	O
whole	O
cell	O
competitive	O
binding	O
assay	O
,	O
normal	B-cell_type
human	I-cell_type
lymphocytes	I-cell_type
had	O
5977	O
+/-	O
1487	O
(	O
mean	O
+/-	O
SD	O
)	O
binding	O
sites/cell	O
and	O
a	O
dissociation	O
constant	O
of	O
10	O
+/-	O
2	O
nM	O
.	O

Lymphocytes	B-cell_type
from	O
patients	O
with	O
untreated	O
adrenal	O
insufficiency	O
had	O
fewer	O
binding	B-protein
sites	I-protein
(	O
3364	O
+/-322	O
)	O
and	O
a	O
2-fold	O
increase	O
in	O
binding	O
affinity	O
(	O
5.4	O
+/-	O
0.9	O
mM	O
)	O
.	O

The	O
administration	O
of	O
conventional	O
replacement	O
doses	O
of	O
cortisone	O
acetate	O
for	O
6	O
months	O
caused	O
no	O
change	O
in	O
receptor	O
number	O
,	O
but	O
was	O
associated	O
with	O
a	O
decrease	O
in	O
binding	O
affinity	O
toward	O
normal	O
.	O

After	O
long	O
term	O
glucocorticoid	O
replacement	O
therapy	O
,	O
binding	O
parameters	O
were	O
similar	O
to	O
those	O
in	O
patients	O
before	O
treatment	O
.	O

The	O
physiological	O
implications	O
of	O
the	O
decreased	O
receptor	O
number	O
and	O
increased	O
binding	O
affinity	O
in	O
adrenal	O
insufficiency	O
remain	O
to	O
be	O
elucidated	O
.	O

-DOCSTART-	O

Glucocorticoid	B-protein
receptor	I-protein
concentrations	O
and	O
terminal	O
transferase	B-protein
activity	O
as	O
indicators	O
of	O
prognosis	O
in	O
acute	O
non-lymphocytic	O
leukaemia	O
.	O

Activity	O
of	O
terminal	B-protein
deoxynucleotidyl	I-protein
transferase	I-protein
(	O
TdT	B-protein
)	O
,	O
adenosine	B-protein
deaminase	I-protein
,	O
and	O
5'nucleotidase	B-protein
and	O
the	O
cellular	O
concentration	O
of	O
glucocorticoid	B-protein
(	I-protein
dexamethasone	I-protein
)	I-protein
receptor	I-protein
were	O
determined	O
in	O
25	O
patients	O
with	O
acute	O
non-lymphocytic	O
leukaemia	O
.	O

All	O
patients	O
were	O
treated	O
according	O
to	O
a	O
common	O
protocol	O
.	O

Increased	O
activity	O
of	O
TdT	B-protein
(	O
greater	O
than	O
0.1	O
unit/microgram	O
DNA	O
)	O
was	O
found	O
in	O
11	O
patients	O
.	O

This	O
group	O
of	O
patients	O
was	O
shown	O
to	O
have	O
higher	O
remission	O
and	O
survival	O
rates	O
(	O
p	O
=	O
0.06	O
)	O
compared	O
with	O
patients	O
with	O
low	O
activity	O
of	O
TdT	O
.	O

The	O
glucocorticoid	B-protein
receptor	I-protein
concentration	O
of	O
the	O
leukaemic	B-cell_type
blast	I-cell_type
cells	I-cell_type
ranged	O
from	O
0	O
to	O
0.94	O
fmol/microgram	O
DNA	O
.	O

Thirteen	O
patients	O
had	O
blast	B-cell_type
cells	I-cell_type
with	O
a	O
glucocorticoid	B-protein
receptor	I-protein
concentration	O
over	O
0.22	O
fmol/microgram	O
DNA	O
.	O

These	O
patients	O
had	O
significantly	O
increased	O
remission	O
and	O
survival	O
rates	O
(	O
p	O
=	O
0.006	O
)	O
compared	O
with	O
those	O
with	O
a	O
low	O
receptor	O
concentration	O
.	O

This	O
finding	O
can	O
not	O
be	O
explained	O
by	O
a	O
difference	O
in	O
sensitivity	O
to	O
glucocorticoids	O
since	O
these	O
were	O
not	O
used	O
as	O
therapeutic	O
agents	O
.	O

Adenosine	B-protein
deaminase	I-protein
and	O
5'nucleotidase	B-protein
activities	O
both	O
varied	O
within	O
two	O
orders	O
of	O
magnitude	O
.	O

No	O
correlation	O
could	O
be	O
found	O
between	O
activities	O
of	O
these	O
enzymes	O
and	O
remission	O
or	O
survival	O
rate	O
.	O

These	O
results	O
show	O
that	O
measurements	O
of	O
TdT	B-protein
activity	O
and	O
the	O
glucocorticoid	B-protein
receptor	I-protein
concentration	O
yield	O
valuable	O
prognostic	O
information	O
in	O
acute	O
non-lymphocytic	O
leukaemia	O

-DOCSTART-	O

[	O
3H	O
]	O
cortivazol	O
:	O
a	O
unique	O
high	O
affinity	O
ligand	O
for	O
the	O
glucocorticoid	B-protein
receptor	I-protein
.	O

Cortivazol	O
(	O
CVZ	O
)	O
and	O
deacylcortivazol	O
(	O
DAC	O
)	O
are	O
pyrazolosteroids	O
with	O
potent	O
glucocorticoid	O
activity	O
.	O

In	O
previous	O
work	O
we	O
showed	O
that	O
DAC	O
is	O
40-fold	O
more	O
potent	O
than	O
dexamethasone	O
(	O
DEX	O
)	O
in	O
lysing	O
leukemic	B-cell_type
lymphoblasts	I-cell_type
.	O

To	O
assess	O
the	O
interaction	O
between	O
these	O
atypical	O
steroids	O
and	O
the	O
glucocorticoid	B-protein
receptor	I-protein
,	O
we	O
examined	O
the	O
binding	O
of	O
[	O
3H	O
]	O
CVZ	O
to	O
cytosol	O
from	O
glucocorticoid-sensitive	B-cell_line
and	I-cell_line
-resistant	I-cell_line
variants	I-cell_line
of	O
the	O
human	B-cell_line
leukemic	I-cell_line
cell	I-cell_line
line	I-cell_line
CEM	B-cell_line
C7	I-cell_line
.	O

In	O
glucocorticoid-sensitive	B-cell_line
cells	I-cell_line
[	O
3H	O
]	O
CVZ	O
causes	O
a	O
2-fold	O
induction	O
of	O
glutamine	B-protein
synthetase	I-protein
and	O
binds	O
to	O
a	O
protein	O
in	O
the	O
4.6	O
S	O
region	O
of	O
high	O
salt	O
sucrose	O
gradients	O
.	O

On	O
DEAE-cellulose	O
chromatography	O
,	O
[	B-protein
3H	I-protein
]	I-protein
CVZ-receptor	I-protein
complexes	I-protein
show	O
a	O
shift	O
from	O
high	O
(	O
0.25	O
M	O
KP	O
)	O
to	O
low	O
salt	O
(	O
0.09	O
M	O
KP	O
)	O
eluting	O
forms	O
upon	O
activation	O
.	O

CVZ	O
competes	O
for	O
a	O
97	O
,	O
000-dalton	O
protein	O
labeled	O
by	O
[	O
3H	O
]	O
dexamethasone	O
mesylate	O
.	O

Scatchard	O
analysis	O
of	O
the	O
binding	O
of	O
[	O
3H	O
]	O
CVZ	O
in	O
glucocorticoid-sensitive	B-cell_line
cells	I-cell_line
revealed	O
a	O
curvilinear	O
plot	O
which	O
resolved	O
into	O
high	O
(	O
0.4	O
nM	O
)	O
and	O
low	O
(	O
11	O
nM	O
)	O
affinity	O
components	O
.	O

The	O
receptor	O
concentration	O
of	O
the	O
low	O
affinity	O
site	O
(	O
0.30	O
pmol/mg	O
protein	O
)	O
was	O
approximately	O
twice	O
that	O
of	O
the	O
high	B-protein
affinity	I-protein
site	I-protein
(	O
0.14	O
pmol/mg	O
protein	O
)	O
.	O

Dissociation	O
experiments	O
with	O
dilution	O
and/or	O
excess	O
unlabeled	O
CVZ	O
supported	O
the	O
presence	O
of	O
independent	O
sites	O
.	O

In	O
contrast	O
,	O
the	O
binding	O
of	O
[	O
3H	O
]	O
DEX	O
to	O
C7	O
cytosol	O
revealed	O
a	O
single	O
class	O
of	O
binding	O
sites	O
(	O
Kd	O
=	O
1.9	O
nM	O
;	O
receptor	O
concentration	O
,	O
0.46	O
pmol/mg	O
protein	O
)	O
.	O

Examination	O
of	O
the	O
binding	O
of	O
[	O
3H	O
]	O
CVZ	O
using	O
10	O
(	O
-5	O
)	O
M	O
DEX	O
as	O
the	O
competing	O
ligand	O
showed	O
that	O
DEX	O
binds	O
only	O
to	O
the	O
low	O
affinity	O
site	O
detected	O
by	O
[	O
3H	O
]	O
CVZ	O
.	O

In	O
cytosol	O
from	O
a	O
glucocorticoid-resistant	B-cell_line
cell	I-cell_line
line	I-cell_line
with	O
virtually	O
no	O
[	O
3H	O
]	O
DEX	O
binding	O
,	O
[	O
3H	O
]	O
CVZ	O
detected	O
a	O
single	O
high	B-protein
affinity	I-protein
binding	I-protein
site	I-protein
that	O
was	O
similar	O
in	O
dissociation	O
constant	O
(	O
0.8	O
nM	O
)	O
and	O
receptor	O
concentration	O
(	O
0.13	O
pmol/mg	O
protein	O
)	O
to	O
the	O
high	B-protein
affinity	I-protein
site	I-protein
detected	O
in	O
the	O
glucocorticoid-sensitive	B-cell_line
cell	I-cell_line
line	I-cell_line
C7	B-cell_line
.	O

-DOCSTART-	O

A	O
controlled	O
pore	O
glass	O
bead	O
assay	O
for	O
the	O
measurement	O
of	O
cytoplasmic	B-protein
and	I-protein
nuclear	I-protein
glucocorticoid	I-protein
receptors	I-protein
.	O

An	O
assay	O
for	O
the	O
quantitation	O
of	O
cytoplasmic	B-protein
and	I-protein
nuclear	I-protein
glucocorticoid	I-protein
receptors	I-protein
in	O
lymphoid	O
tissue	O
has	O
been	O
developed	O
using	O
controlled	O
pore	O
glass	O
(	O
CPG	O
)	O
beads	O
.	O

Soluble	O
receptor	B-protein
--	I-protein
3H-steroid	I-protein
complex	I-protein
(	O
cytosol	O
or	O
nuclear	O
extract	O
)	O
is	O
adsorbed	O
quantitatively	O
within	O
the	O
crevasses	O
of	O
porous	O
glass	O
beads	O
.	O

Excess	O
labeled	O
steroid	O
as	O
well	O
as	O
most	O
non-specifically	O
bound	O
steroid	O
is	O
easily	O
washed	O
away	O
,	O
leaving	O
the	O
hormone-receptor	B-protein
complex	I-protein
retained	O
by	O
the	O
beads	O
.	O

Bound	O
3H-steroid	O
is	O
eluted	O
with	O
ethanol	O
and	O
measured	O
for	O
radioactivity	O
.	O

This	O
procedure	O
which	O
is	O
simple	O
,	O
rapid	O
,	O
and	O
highly	O
reproducible	O
is	O
carried	O
out	O
using	O
frozen	O
samples	O
(	O
stable	O
for	O
many	O
months	O
)	O
containing	O
as	O
few	O
as	O
1	O
X	O
10	O
(	O
7	O
)	O
cells	O
.	O

A	O
comparison	O
of	O
the	O
CPG	O
assay	O
to	O
dextran	O
coated	O
charcoal	O
and	O
a	O
whole	O
cell	O
assay	O
demonstrates	O
that	O
CPG	O
and	O
dextran	O
coated	O
charcoal	O
give	O
equivalent	O
measurements	O
of	O
cytosolic	B-protein
receptor	I-protein
concentration	O
,	O
while	O
the	O
CPG	O
and	O
whole	O
cell	O
assays	O
provide	O
equivalent	O
values	O
for	O
total	O
receptor	O
content	O
.	O

-DOCSTART-	O

Plasmacytoid	O
blast	O
crisis	O
in	O
B-cell	B-cell_type
chronic	O
lymphocytic	O
leukemia	O
:	O
effect	O
of	O
estradiol	O
on	O
growth	O
and	O
differentiation	O
in	O
vitro	O
.	O

Evolution	O
of	O
a	O
case	O
of	O
chronic	O
lymphocytic	O
leukemia	O
(	O
CLL	O
)	O
into	O
blast	O
crisis	O
was	O
found	O
to	O
be	O
characterized	O
by	O
three	O
unusual	O
features	O
(	O
1	O
)	O
the	O
phenotype	O
of	O
the	O
emerging	B-cell_type
blast	I-cell_type
cells	I-cell_type
was	O
that	O
of	O
pre-plasmacytoid	B-cell_type
cells	I-cell_type
as	O
shown	O
by	O
plasma	O
cell	O
morphology	O
and	O
an	O
immunological	O
phenotype	O
corresponding	O
partially	O
with	O
CLL-	B-cell_type
or	I-cell_type
intermediate	I-cell_type
B-cells	I-cell_type
,	O
partially	O
with	O
plasma	B-cell_type
cells	I-cell_type
(	O
terminal	B-protein
transferase	I-protein
-	O
,	O
common	O
acute	B-protein
lymphocytic	I-protein
leukemia	I-protein
antigen	I-protein
-	O
,	O
Ia+	B-protein
,	O
surface	B-protein
immunoglobulin	I-protein
heavy	I-protein
chains	I-protein
-	O
,	O
surface	B-protein
kappa	I-protein
light	I-protein
chains	I-protein
+	O
,	O
intracytoplasmic	B-protein
immunoglobulin	I-protein
A+	I-protein
and	I-protein
G+	I-protein
,	O
BA-1+	B-protein
,	O
polyclonal	O
gammaglobulin	O
production	O
)	O
;	O
(	O
2	O
)	O
cytogenetic	O
analysis	O
of	O
spontaneous	O
metaphases	O
revealed	O
that	O
in	O
addition	O
to	O
the	O
typical	O
CLL	O
abnormality	O
,	O
trisomy	O
12	O
,	O
in	O
all	O
of	O
the	O
cells	O
,	O
an	O
additional	O
translocation	O
between	O
chromosomes	B-DNA
14	I-DNA
and	I-DNA
17	I-DNA
was	O
present	O
in	O
40	O
%	O
with	O
a	O
presumptive	O
breakpoint	O
on	O
chromosome	B-DNA
14	I-DNA
(	O
q12-3	O
)	O
never	O
described	O
before	O
(	O
commonly	O
q32	O
)	O
and	O
(	O
3	O
)	O
the	O
progression	O
of	O
the	O
disease	O
was	O
associated	O
with	O
a	O
striking	O
increase	O
in	O
the	O
expression	O
by	O
the	O
transformed	B-cell_type
cells	I-cell_type
of	O
specific	O
binding	O
sites	O
for	O
estradiol	O
(	O
E2	O
)	O
due	O
to	O
an	O
actual	O
increase	O
in	O
total	O
cellular	B-protein
receptor	I-protein
proteins	I-protein
and	O
not	O
to	O
a	O
change	O
in	O
receptor	O
affinity	O
for	O
E2	O
.	O

The	O
functional	O
status	O
of	O
the	O
steroid	B-protein
receptors	I-protein
was	O
confirmed	O
by	O
nuclear	O
transfer	O
of	O
the	O
cytoplasmic	B-protein
hormone-receptor	I-protein
complex	I-protein
upon	O
temperature	O
activation	O
.	O

Since	O
the	O
rise	O
in	O
E2-receptor	B-protein
display	O
paralleled	O
a	O
large	O
increase	O
in	O
the	O
proliferative	O
activity	O
of	O
the	O
cells	O
as	O
well	O
as	O
a	O
change	O
in	O
their	O
maturation	O
status	O
the	O
question	O
was	O
raised	O
as	O
to	O
whether	O
the	O
E2-receptor	B-protein
should	O
be	O
considered	O
as	O
a	O
physiological	O
marker	O
of	O
growth	O
rate	O
or	O
of	O
cellular	O
differentiation	O
.	O

Exposure	O
of	O
the	O
patient	O
's	O
blast	B-cell_type
cells	I-cell_type
to	O
E2	O
in	O
vitro	O
resulted	O
in	O
cessation	O
of	O
cell	O
growth	O
following	O
at	O
least	O
one	O
mitosis	O
after	O
addition	O
of	O
the	O
inducer	O
as	O
seen	O
from	O
the	O
replacement	O
of	O
the	O
large	B-cell_type
blasts	I-cell_type
by	O
small	O
CLL-like	B-cell_type
cells	I-cell_type
without	O
definite	O
signs	O
of	O
alteration	O
of	O
the	O
differentiation	O
status	O
.	O

This	O
suggests	O
the	O
association	O
of	O
E2-receptor	B-protein
expression	O
with	O
control	O
of	O
growth	O
rather	O
than	O
cell	O
maturation	O
.	O

-DOCSTART-	O

Multiple	O
forms	O
and	O
fragments	O
of	O
cytosolic	B-protein
glucocorticoid	I-protein
receptors	I-protein
from	O
human	B-cell_type
leukemic	I-cell_type
cells	I-cell_type
and	O
normal	B-cell_type
lymphocytes	I-cell_type
.	O

Therapy	O
with	O
glucocorticoids	O
is	O
generally	O
more	O
effective	O
in	O
acute	O
lymphoblastic	O
leukemia	O
than	O
in	O
other	O
types	O
of	O
human	O
leukemia	O
.	O

Previous	O
studies	O
,	O
however	O
,	O
have	O
not	O
revealed	O
any	O
consistent	O
relationship	O
between	O
clinical	O
responsiveness	O
and	O
the	O
cellular	O
or	O
cytosolic	O
concentration	O
of	O
glucocorticoid-binding	B-protein
sites	I-protein
.	O

The	O
objectives	O
of	O
this	O
study	O
were	O
to	O
determine	O
whether	O
there	O
are	O
intrinsic	O
structural	O
differences	O
among	O
the	O
glucocorticoid	B-protein
receptors	I-protein
in	O
various	O
types	O
of	O
leukemic	B-cell_type
cells	I-cell_type
and	O
normal	B-cell_type
lymphocytes	I-cell_type
and	O
to	O
investigate	O
the	O
role	O
of	O
endogenous	B-protein
peptidases	I-protein
in	O
receptor	O
degradation	O
.	O

Cytosols	O
were	O
prepared	O
from	O
fresh	B-cell_type
or	I-cell_type
rapidly	I-cell_type
frozen	I-cell_type
leukocytes	I-cell_type
from	O
6	O
healthy	O
adults	O
and	O
35	O
high-risk	O
leukemia	O
patients	O
(	O
median	O
white	O
blood	O
cell	O
count	O
,	O
150	O
,	O
000	O
cells/microliter	O
;	O
median	O
age	O
,	O
13	O
years	O
)	O
.	O

Receptors	O
were	O
labeled	O
with	O
[	O
3H	O
]	O
triamcinolone	O
acetonide	O
and	O
quantitated	O
by	O
charcoal-dextran	O
treatment	O
or	O
Sephadex	O
LH-20	O
chromatography	O
.	O

Mean	O
and	O
median	O
cytosolic	O
receptor	O
concentrations	O
in	O
12	O
acute	O
lymphoblastic	O
leukemia	O
specimens	O
lacking	O
the	O
standard	O
B-cell	O
or	O
T-cell	O
markers	O
(	O
``	O
null	O
cells	O
''	O
)	O
were	O
approximately	O
4-fold	O
higher	O
than	O
in	O
23	O
other	O
leukemic	B-cell_type
cell	I-cell_type
specimens	I-cell_type
.	O

No	O
other	O
consistent	O
differences	O
in	O
receptor	O
content	O
were	O
observed	O
.	O

Agarose	O
filtration	O
and	O
ultracentrifugation	O
in	O
hypotonic	O
buffers	O
containing	O
20	O
mM	O
Na2MoO4	O
revealed	O
complexes	O
of	O
similar	O
size	O
and	O
shape	O
in	O
all	O
clinical	O
specimens	O
tested	O
and	O
two	O
established	O
leukemic	B-cell_line
cell	I-cell_line
lines	I-cell_line
.	O

They	O
had	O
Stokes	O
radii	O
(	O
Rs	O
)	O
of	O
8.1	O
+/-	O
0.5	O
(	O
S.D.	O
)	O
nm	O
(	O
n	O
=	O
50	O
)	O
,	O
sedimentation	O
coefficients	O
of	O
9.5	O
+/-	O
0.3S	O
(	O
n	O
=	O
40	O
)	O
,	O
molecular	O
weights	O
of	O
approximately	O
330	O
,	O
000	O
,	O
and	O
axial	O
ratios	O
(	O
a/b	O
)	O
of	O
approximately	O
12	O
.	O

In	O
hypertonic	O
,	O
molybdate-free	O
buffer	O
,	O
these	O
oligomeric	B-protein
complexes	I-protein
were	O
dissociated	O
into	O
subunits	O
with	O
Rs	O
of	O
5.9	O
+/-	O
0.3	O
nm	O
(	O
n	O
=	O
12	O
)	O
and	O
a/b	O
of	O
11	O
to	O
12	O
,	O
as	O
observed	O
previously	O
for	O
other	O
receptors	O
.	O

Fragmentation	O
of	O
the	O
oligomer	B-protein
and	O
the	O
subunit	B-protein
was	O
evident	O
in	O
some	O
cytosols	O
.	O

High	O
activities	O
of	O
peptidases	B-protein
of	O
various	O
specificities	O
were	O
detected	O
in	O
leukemic	O
cell	O
cytosols	O
,	O
as	O
in	O
other	O
cytosols	O
,	O
by	O
fluorometric	O
assays	O
with	O
derivatives	O
of	O
7-amino-4-methylcoumarin	O
.	O

Receptor	O
cleavage	O
by	O
these	O
and	O
other	O
endogenous	B-protein
enzymes	I-protein
may	O
account	O
for	O
previous	O
observations	O
of	O
``	O
abnormal	O
''	O
receptors	O
in	O
cytosols	O
from	O
some	O
leukemic	B-cell_type
specimens	I-cell_type
.	O

We	O
conclude	O
that	O
intrinsic	O
structural	O
defects	O
in	O
the	O
receptors	O
are	O
unlikely	O
explanations	O
for	O
the	O
unresponsiveness	O
of	O
some	O
types	O
of	O
leukemia	O
to	O
steroid	O
therapy	O
.	O

-DOCSTART-	O

Glucocorticoid	B-protein
receptor	I-protein
number	O
and	O
intracellular	O
water	O
space	O
.	O

In	O
order	O
to	O
elucidate	O
the	O
relationship	O
between	O
cell	O
water	O
content	O
and	O
number	O
of	O
glucocorticoid	B-protein
receptors	I-protein
,	O
eleven	O
normal	O
and	O
malignant	B-cell_type
lymphoid	I-cell_type
or	I-cell_type
myelomonocytic	I-cell_type
cell	I-cell_type
types	I-cell_type
originating	O
from	O
mouse	O
,	O
rat	O
and	O
man	O
were	O
investigated	O
.	O

The	O
cellular	O
water	O
space	O
was	O
measured	O
with	O
3H2O	O
,	O
and	O
glucocorticoid	B-protein
receptor	I-protein
number	O
was	O
measured	O
in	O
a	O
whole-cell	O
binding	O
assay	O
with	O
[	O
3H	O
]	O
dexamethasone	O
at	O
30	O
and	O
37	O
degrees	O
C	O
.	O

The	O
intracellular	O
water	O
phase	O
concentration	O
of	O
glucocorticoid	B-protein
receptors	I-protein
(	O
around	O
40	O
nmol/l	O
cell	O
water	O
)	O
,	O
and	O
the	O
dependence	O
of	O
receptor	O
affinity	O
on	O
temperature	O
were	O
similar	O
in	O
normal	B-cell_type
and	I-cell_type
malignant	I-cell_type
rodent	I-cell_type
and	I-cell_type
human	I-cell_type
cells	I-cell_type
.	O

It	O
is	O
concluded	O
that	O
comparisons	O
of	O
glucocorticoid	B-protein
receptor	I-protein
levels	O
are	O
best	O
made	O
on	O
the	O
basis	O
of	O
intracellular	B-protein
receptor	I-protein
concentrations	O
.	O

-DOCSTART-	O

Covalent	O
labeling	O
of	O
rat	B-cell_type
thymocyte	I-cell_type
and	O
human	B-protein
lymphoid	I-protein
glucocorticoid	I-protein
receptor	I-protein
.	O

Lymphoid	B-cell_type
cells	I-cell_type
contain	O
specific	O
receptors	B-protein
for	O
glucocorticoids	O
.	O

We	O
have	O
used	O
[	O
3H	O
]	O
dexamethasone-21-mesylate	O
to	O
label	O
covalently	O
glucocorticoid	B-protein
receptors	I-protein
in	O
rat	B-cell_type
thymic	I-cell_type
lymphocytes	I-cell_type
and	O
in	O
neoplastic	B-cell_type
cells	I-cell_type
obtained	O
from	O
patients	O
with	O
acute	O
lymphoblastic	O
leukemia	O
and	O
malignant	O
lymphoma	O
.	O

The	O
covalently	O
labeled	O
glucocorticoid	B-protein
receptors	I-protein
were	O
identified	O
by	O
polyacrylamide	O
gel	O
electrophoresis	O
(	O
in	O
the	O
presence	O
of	O
0.1	O
%	O
sodium	O
dodecyl	O
sulfate	O
)	O
.	O

In	O
cytosolic	O
fractions	O
prepared	O
from	O
rat	B-cell_type
thymic	I-cell_type
lymphocytes	I-cell_type
,	O
[	O
3H	O
]	O
-dexamethasone-21-mesylate	O
labels	O
a	O
protein	O
(	O
Mr	O
approximately	O
equal	O
to	O
95	O
,	O
000	O
)	O
which	O
was	O
identified	O
as	O
the	O
glucocorticoid	B-protein
receptor	I-protein
by	O
the	O
following	O
criteria	O
:	O
(	O
a	O
)	O
labeling	O
of	O
this	O
moiety	O
is	O
inhibited	O
by	O
treatment	O
with	O
a	O
100-fold	O
molar	O
excess	O
of	O
glucocorticoids	O
,	O
such	O
as	O
dexamethasone	O
and	O
triamcinolone	O
acetonide	O
;	O
and	O
(	O
b	O
)	O
the	O
covalently	O
labeled	O
Mr	O
approximately	O
equal	O
to	O
95	O
,	O
000	O
protein	O
is	O
activated	O
(	O
by	O
heating	O
at	O
20	O
degrees	O
for	O
30	O
min	O
)	O
to	O
a	O
form	O
that	O
binds	O
to	O
DNA-cellulose	O
.	O

When	O
intact	B-cell_type
thymocytes	I-cell_type
are	O
treated	O
with	O
[	O
3H	O
]	O
dexamethasone-21-mesylate	O
,	O
an	O
Mr	O
approximately	O
equal	O
to	O
95	O
,	O
000	O
moiety	O
is	O
also	O
labeled	O
covalently	O
.	O

Approximately	O
35	O
%	O
of	O
the	O
glucocorticoid	B-protein
receptors	I-protein
can	O
be	O
labeled	O
covalently	O
when	O
intact	O
thymocytes	B-cell_type
are	O
treated	O
with	O
100	O
nM	O
[	O
3H	O
]	O
dexamethasone-21-mesylate	O
for	O
30	O
min	O
at	O
4	O
degrees	O
.	O

Neoplastic	B-cell_type
cells	I-cell_type
from	O
acute	O
lymphoblastic	O
leukemia	O
and	O
malignant	O
lymphoma	O
were	O
treated	O
with	O
[	O
3H	O
]	O
dexamethasone-21-mesylate	O
.	O

In	O
all	O
samples	O
,	O
an	O
Mr	O
approximately	O
equal	O
to	O
95	O
,	O
000	O
moiety	O
was	O
labeled	O
covalently	O
;	O
labeling	O
was	O
inhibited	O
by	O
excess	O
glucocorticoid	O
.	O

Smaller	B-protein
moieties	I-protein
were	O
also	O
identified	O
by	O
competition	O
experiments	O
;	O
these	O
may	O
represent	O
proteolytic	B-protein
fragments	I-protein
of	O
the	O
Mr	O
approximately	O
equal	O
to	O
95	O
,	O
000	O
receptor	O
.	O

Thus	O
,	O
in	O
rat	B-cell_type
and	I-cell_type
human	I-cell_type
lymphoid	I-cell_type
cells	I-cell_type
,	O
[	O
3H	O
]	O
dexamethasone-21-mesylate	O
can	O
be	O
used	O
to	O
label	O
covalently	O
the	O
glucocorticoid	B-protein
receptor	I-protein
.	O

-DOCSTART-	O

The	O
association	O
of	O
cytosol	O
oestrogen	B-protein
and	I-protein
progesterone	I-protein
receptors	I-protein
with	O
histological	O
features	O
of	O
breast	O
cancer	O
and	O
early	O
recurrence	O
of	O
disease	O
.	O

Two	O
hundred	O
and	O
eighty-eight	O
primary	O
breast	O
tumours	O
were	O
examined	O
for	O
the	O
presence	O
or	O
absence	O
of	O
oestrogen	B-protein
(	I-protein
REc	I-protein
)	I-protein
and	I-protein
progesterone	I-protein
(	I-protein
RPc	I-protein
)	I-protein
receptors	I-protein
.	O

Analysis	O
has	O
shown	O
a	O
relative	O
interdependence	O
between	O
the	O
steroid	B-protein
receptor	I-protein
status	O
of	O
primary	O
breast	O
cancer	O
and	O
other	O
prognostic	O
variables	O
such	O
as	O
histological	O
grade	O
,	O
lymphocytic	O
infiltration	O
and	O
tumour	O
elastosis	O
.	O

There	O
were	O
significant	O
associations	O
between	O
epithelial	O
cellularity	O
,	O
stromal	O
fibrosis	O
and	O
the	O
value	O
of	O
REc	B-protein
in	O
those	O
tumours	O
in	O
which	O
the	O
receptor	O
was	O
present	O
.	O

Cellularity	O
and	O
fibrosis	O
were	O
unrelated	O
to	O
the	O
presence	O
or	O
absence	O
of	O
oestrogen	B-protein
receptor	I-protein
.	O

By	O
contrast	O
,	O
neither	O
the	O
presence	O
or	O
absence	O
nor	O
the	O
value	O
of	O
RPc	B-protein
could	O
be	O
related	O
to	O
cellularity	O
or	O
fibrosis	O
.	O

The	O
value	O
of	O
REc	B-protein
and	O
RPc	B-protein
analysis	O
as	O
an	O
indicator	O
of	O
prognosis	O
was	O
examined	O
in	O
a	O
sub-group	O
of	O
175	O
patients	O
receiving	O
no	O
additional	O
treatment	O
following	O
mastectomy	O
.	O

Overall	O
relapse-free	O
survival	O
(	O
RFS	O
)	O
was	O
no	O
different	O
for	O
those	O
patients	O
with	O
receptors	O
compared	O
to	O
those	O
without	O
them	O
(	O
REc	B-protein
P	O
=	O
0.11	O
,	O
RPc	O
P	O
=	O
0.7	O
)	O
.	O

There	O
was	O
no	O
difference	O
in	O
RFS	O
of	O
receptor	O
positive	O
and	O
negative	O
tumours	O
when	O
the	O
axillary	O
node	O
status	O
was	O
taken	O
into	O
account	O
.	O

-DOCSTART-	O

Clinical	O
implications	O
of	O
glucocorticoid	B-protein
receptors	I-protein
in	O
human	O
leukemia	O
.	O

Normal	B-cell_type
lymphoid	I-cell_type
cells	I-cell_type
contain	O
glucocorticoid	B-protein
receptor	I-protein
.	O

A	O
variety	O
of	O
stimuli	O
that	O
activate	O
these	O
cells	O
also	O
induce	O
increases	O
in	O
receptor	O
concentration	O
.	O

Similar	O
glucocorticoid	B-protein
receptors	I-protein
can	O
be	O
detected	O
in	O
lymphoid	B-cell_type
cells	I-cell_type
from	O
patients	O
with	O
acute	O
lymphoblastic	O
leukemia	O
(	O
ALL	O
)	O
.	O

Absence	O
of	O
the	O
glucocorticoid	B-protein
receptor	I-protein
(	O
usually	O
found	O
in	O
treated	O
patients	O
)	O
predicts	O
lack	O
of	O
glucocorticoid	O
responsiveness	O
.	O

Furthermore	O
,	O
in	O
our	O
hands	O
,	O
glucocorticoid	B-protein
receptor	I-protein
levels	O
correlate	O
with	O
the	O
duration	O
of	O
complete	O
remission	O
in	O
ALL	O
(	O
though	O
not	O
in	O
other	O
forms	O
of	O
leukemia	O
)	O
.	O

This	O
association	O
is	O
independent	O
of	O
cell	O
type	O
,	O
age	O
,	O
sex	O
,	O
or	O
initial	O
leukocyte	B-cell_type
count	O
.	O

The	O
level	O
of	O
receptor	O
shows	O
a	O
negative	O
correlation	O
with	O
increasing	O
aggressiveness	O
of	O
the	O
tumor	O
(	O
null-cell	O
leukemia	O
greater	O
than	O
T-cell	O
leukemia	O
greater	O
than	O
Burkitt	O
's	O
lymphoma	O
)	O
.	O

-DOCSTART-	O

Corticosteroid-mediated	O
immunoregulation	O
in	O
man	O
.	O

Glucocorticoids	O
have	O
profound	O
and	O
complex	O
effects	O
on	O
the	O
human	O
immune	O
response	O
.	O

However	O
,	O
the	O
precise	O
mechanisms	O
of	O
the	O
corticosteroid-induced	O
immunoregulation	O
in	O
man	O
have	O
not	O
been	O
precisely	O
defined	O
.	O

Intracytoplasmic	B-protein
corticosteroid-specific	I-protein
receptors	I-protein
appear	O
to	O
be	O
an	O
important	O
common	O
pathway	O
for	O
steroid-induced	O
changes	O
,	O
but	O
variations	O
of	O
receptor	O
parameters	O
do	O
not	O
account	O
for	O
the	O
multifaceted	O
effects	O
on	O
the	O
immune	O
system	O
.	O

Human	B-cell_type
circulating	I-cell_type
mononuclear	I-cell_type
cells	I-cell_type
redistribute	O
out	O
of	O
the	O
intravascular	O
compartment	O
following	O
treatment	O
with	O
corticosteroids	O
.	O

Although	O
certain	O
components	O
at	O
this	O
redistribution	O
phenomenon	O
have	O
been	O
well-characterized	O
,	O
the	O
importance	O
of	O
this	O
compartmental	O
cellular	O
shift	O
with	O
respect	O
to	O
the	O
mechanisms	O
of	O
corticosteroid-induced	O
immunoregulation	O
are	O
less	O
well-defined	O
.	O

Recent	O
observations	O
that	O
activated	B-cell_type
lymphocytes	I-cell_type
may	O
be	O
sensitive	O
to	O
the	O
lytic	O
effects	O
of	O
glucocorticoids	O
suggest	O
that	O
under	O
certain	O
situations	O
the	O
elimination	O
of	O
selected	O
subsets	O
of	O
cells	O
may	O
be	O
a	O
relevant	O
mechanism	O
of	O
corticosteroid-mediated	O
immunoregulation	O
in	O
man	O
.	O

Corticosteroid-mediated	O
effects	O
on	O
monocyte	B-cell_type
function	O
may	O
be	O
an	O
important	O
mechanism	O
of	O
drug-induced	B-cell_type
immunoregulation	I-cell_type
in	O
monocyte-dependent	B-cell_type
responses	I-cell_type
.	O

In	O
some	O
experimental	O
conditions	O
,	O
corticosteroids	O
inhibit	O
Interleukin	B-protein
1	I-protein
production	O
by	O
monocytes	B-cell_type
.	O

The	O
immunoregulatory	O
effects	O
of	O
corticosteroids	O
on	O
lymphocyte	B-cell_type
immune	O
responses	O
are	O
complex	O
.	O

In	O
vitro	O
corticosteroids	O
appear	O
to	O
selectively	O
affect	O
early	O
immunoregulatory	O
events	O
as	O
opposed	O
to	O
altering	O
an	O
established	O
response	O
.	O

Multiple	O
sites	O
of	O
steroid-induced	O
modulations	O
of	O
human	B-cell_type
B	I-cell_type
cell	I-cell_type
responses	O
have	O
been	O
defined	O
.	O

-DOCSTART-	O

Human	B-cell_line
lymphoid	I-cell_line
cell	I-cell_line
lines	I-cell_line
and	O
glucocorticoids	O
:	O
II	O
.	O

Whole	O
cell	O
and	O
cytoplasmic	O
binding	O
properties	O
of	O
lymphoblastoid	B-cell_line
,	I-cell_line
leukaemia	I-cell_line
and	I-cell_line
lymphoma	I-cell_line
lines	I-cell_line
.	O

The	O
glucocorticoid	O
binding	O
properties	O
of	O
18	O
human	B-cell_line
lymphoid	I-cell_line
cell	I-cell_line
lines	I-cell_line
(	O
HLCL	B-cell_line
)	O
have	O
been	O
investigated	O
.	O

The	O
specificity	O
of	O
steroid	O
binding	O
was	O
confirmed	O
with	O
various	O
glucocorticoid	O
agonists	O
and	O
antagonists	O
.	O

A	O
gradation	O
in	O
whole	O
cell	O
and	O
cytoplasmic	O
glucocorticoid	O
binding	O
capacity	O
was	O
observed	O
in	O
the	O
different	O
cell	O
line	O
types	O
:	O
lymphoblastoid	B-cell_line
greater	O
than	O
lymphoma	B-cell_line
greater	O
than	O
leukaemia	B-cell_line
.	O

The	O
cytoplasmic	B-protein
receptors	I-protein
of	O
leukaemia	B-cell_line
and	I-cell_line
lymphoblastoid	I-cell_line
lines	I-cell_line
appeared	O
to	O
contain	O
both	O
proteinaceous	O
and	O
phospholipid	O
components	O
.	O

Cytoplasmic	B-protein
steroid-receptor	I-protein
complexes	I-protein
exhibited	O
a	O
wide	O
range	O
of	O
sedimentation	O
coefficients	O
(	O
8.5-11.3S	O
)	O
in	O
low	O
ionic	O
strength	O
buffer	O
but	O
there	O
was	O
no	O
correlation	O
with	O
cell	O
line	O
type	O
or	O
glucocorticoid	O
sensitivity	O
.	O

Activation	O
of	O
these	O
complexes	O
by	O
heat	O
(	O
37	O
degrees	O
C	O
)	O
or	O
exposure	O
to	O
high	O
ionic	O
strength	O
buffer	O
(	O
0.3	O
M	O
NaCl	O
)	O
induced	O
nuclear	O
binding	O
of	O
steroid	O
but	O
only	O
complexes	O
in	O
high	O
ionic	O
strength	O
buffer	O
manifested	O
changes	O
in	O
sedimentation	O
coefficient	O
.	O

No	O
correlation	O
was	O
observed	O
between	O
the	O
level	O
or	O
nature	O
of	O
glucocorticoid	O
binding	O
and	O
the	O
cytolethal	O
or	O
cytostatic	O
responsiveness	O
of	O
HLCL	B-cell_line
to	O
glucocorticoid	O
treatment	O
in	O
vitro	O
.	O

The	O
resistance	O
to	O
cytolethal	O
effects	O
can	O
not	O
be	O
ascribed	O
to	O
a	O
failure	O
of	O
cells	O
to	O
take	O
up	O
and	O
bind	O
steroid	O
or	O
to	O
significant	O
differences	O
in	O
the	O
molecular	O
species	O
of	O
cytoplasmic	B-protein
receptors	I-protein
present	O
.	O

The	O
molecular	O
mechanisms	O
by	O
which	O
glucocorticoids	O
achieve	O
cytolethal	O
responses	O
in	O
human	B-cell_type
lymphoid	I-cell_type
cells	I-cell_type

-DOCSTART-	O

CD28	B-protein
-mediated	O
activation	O
in	O
CD45RA+	B-cell_type
and	I-cell_type
CD45RO+	I-cell_type
T	I-cell_type
cells	I-cell_type
:	O
enhanced	O
levels	O
of	O
reactive	O
oxygen	O
intermediates	O
and	O
c-Rel	B-protein
nuclear	O
translocation	O
in	O
CD45RA+	B-cell_type
cells	I-cell_type
.	O

We	O
have	O
analyzed	O
the	O
effect	O
of	O
complete	O
T	O
cell	O
activation	O
(	O
anti-CD3	B-protein
plus	O
anti-CD28	B-protein
)	O
on	O
the	O
activation	O
of	O
NF-kappaB	B-protein
in	O
CD45RA+	B-cell_type
(	I-cell_type
naive	I-cell_type
)	I-cell_type
and	I-cell_type
CD45RO+	I-cell_type
(	I-cell_type
memory/effector	I-cell_type
)	I-cell_type
T	I-cell_type
cells	I-cell_type
.	O

Long	O
exposure	O
(	O
24	O
h	O
)	O
induced	O
stronger	O
NF-kappaB	B-protein
DNA	O
binding	O
in	O
CD45RA+	B-cell_type
cells	I-cell_type
than	O
in	O
CD45RO+	B-cell_type
cells	I-cell_type
.	O

Analysis	O
of	O
the	O
nuclear	B-protein
c-Rel	I-protein
protein	I-protein
indicated	O
that	O
after	O
anti-CD3+anti-CD28	O
stimulation	O
the	O
level	O
of	O
c-Rel	B-protein
was	O
higher	O
in	O
CD45RA+	B-cell_type
cells	I-cell_type
.	O

Analysis	O
of	O
the	O
cytoplasmic	O
inhibitor	O
IkappaBalpha	B-protein
indicated	O
that	O
anti-CD3+anti-CD28	O
stimulation	O
induced	O
a	O
long-lasting	O
degradation	O
in	O
CD45RA+	B-cell_type
cells	I-cell_type
but	O
in	O
CD45RO+	B-cell_type
cells	I-cell_type
the	O
degradation	O
process	O
was	O
more	O
rapid	O
.	O

Because	O
the	O
CD28	O
costimulus	O
is	O
known	O
to	O
induce	O
the	O
production	O
of	O
reactive	O
oxygen	O
intermediates	O
(	O
ROIs	O
)	O
,	O
the	O
intracellular	O
ROI	O
levels	O
in	O
CD45RA+	B-cell_type
and	I-cell_type
CD45RO+	I-cell_type
cells	I-cell_type
were	O
compared	O
by	O
flow	O
cytometry	O
.	O

ROIs	O
were	O
produced	O
in	O
both	O
cell	O
types	O
,	O
but	O
more	O
strongly	O
in	O
CD45RA+	B-cell_type
cells	I-cell_type
.	O

The	O
data	O
presented	O
in	O
this	O
study	O
further	O
emphasize	O
the	O
differences	O
between	O
CD45RA+	B-cell_type
and	I-cell_type
CD45RO+	I-cell_type
T	I-cell_type
lymphocytes	I-cell_type
in	O
ROI-dependent	O
signaling	O
pathways	O
.	O

-DOCSTART-	O

Ikaros	B-protein
in	O
hemopoietic	B-cell_line
lineage	I-cell_line
determination	O
and	O
homeostasis	O
.	O

Studies	O
on	O
the	O
molecular	O
mechanisms	O
that	O
control	O
hemopoietic	O
differentiation	O
have	O
focused	O
on	O
signaling	O
cascades	O
and	O
nuclear	O
effectors	O
that	O
drive	O
this	O
complex	O
developmental	O
system	O
in	O
a	O
regulated	O
fashion	O
.	O

Here	O
we	O
review	O
the	O
role	O
of	O
Ikaros	B-protein
,	O
the	O
founding	O
member	O
of	O
a	O
unique	O
family	O
of	O
zinc	B-protein
finger	I-protein
transcription	I-protein
factors	I-protein
in	O
this	O
developmental	O
process	O
.	O

Studies	O
on	O
an	O
Ikaros	B-protein
null	O
mutation	O
have	O
revealed	O
an	O
essential	O
role	O
for	O
this	O
factor	O
in	O
lymphoid	B-cell_type
cell	I-cell_type
fate	O
determination	O
and	O
at	O
subsequent	O
branch	O
points	O
of	O
the	O
T	B-cell_type
cell	I-cell_type
differentiation	O
pathway	O
.	O

Differences	O
in	O
the	O
phenotypes	O
of	O
a	O
null	O
and	O
a	O
dominant	O
negative	O
(	O
DN	O
)	O
Ikaros	O
mutation	O
provide	O
insight	O
into	O
a	O
regulatory	O
network	O
through	O
which	O
Ikaros	B-protein
proteins	I-protein
exert	O
their	O
effects	O
in	O
development	O
.	O

In	O
addition	O
a	O
comparative	O
analysis	O
of	O
the	O
hemopoietic	B-cell_type
stem	I-cell_type
cell	I-cell_type
and	O
precursor	O
compartment	O
resulting	O
from	O
the	O
two	O
Ikaros	B-protein
mutations	O
reveals	O
a	O
profound	O
yet	O
not	O
absolute	O
requirement	O
for	O
Ikaros	B-protein
in	O
the	O
production	O
and	O
differentiation	O
of	O
these	O
populations	O
.	O

-DOCSTART-	O

Overexpression	O
of	O
p65	O
and	O
c-Jun	O
substitutes	O
for	O
B7-1	O
costimulation	O
by	O
targeting	O
the	O
CD28RE	B-DNA
within	O
the	O
IL-2	B-DNA
promoter	I-DNA
.	O

The	O
role	O
of	O
Rel	B-protein
and	O
activation	B-protein
protein-1	I-protein
(	O
AP-1	B-protein
)	O
in	O
IL-2	B-DNA
promoter	I-DNA
activity	O
in	O
B7-1-	B-cell_type
and	I-cell_type
leukocyte	I-cell_type
function-associated	I-cell_type
Ag-3	I-cell_type
(	I-cell_type
LFA.	I-cell_type
3	I-cell_type
)	I-cell_type
-costimulated	I-cell_type
T	I-cell_type
cells	I-cell_type
has	O
been	O
evaluated	O
.	O

We	O
demonstrate	O
that	O
overexpression	O
of	O
c-Jun	B-protein
but	O
not	O
c-Fos	B-protein
increases	O
IL-2	B-DNA
promoter	I-DNA
activity	O
in	O
both	O
B7-1-	B-cell_type
and	I-cell_type
LFA-3-costimulated	I-cell_type
Jurkat	I-cell_type
T	I-cell_type
cells	I-cell_type
.	O

Cotransfection	O
of	O
both	O
c-Jun	O
and	O
c-Fos	O
substitutes	O
for	O
B7-1	O
costimulation	O
in	O
driving	O
an	O
activation	B-DNA
protein-1	I-DNA
response	I-DNA
element	I-DNA
but	O
not	O
for	O
the	O
IL-2	B-DNA
promoter	I-DNA
.	O

Overexpression	O
of	O
Rel	B-protein
proteins	I-protein
demonstrated	O
that	O
p65-expressing	B-cell_line
Jurkat	I-cell_line
cells	I-cell_line
transcribed	O
equally	O
well	O
a	O
nuclear	B-DNA
factor	I-DNA
kappabeta	I-DNA
reporter	I-DNA
construct	I-DNA
when	O
costimulated	O
with	O
B7-1	B-protein
or	O
LFA-3	B-protein
,	O
but	O
transcription	O
of	O
IL-2	B-DNA
promoter	I-DNA
or	O
CD28	B-DNA
response	I-DNA
element	I-DNA
(	I-DNA
CD28RE	I-DNA
)	I-DNA
-driven	I-DNA
reporters	I-DNA
was	O
superior	O
in	O
B7-1-costimulated	B-cell_line
cells	I-cell_line
.	O

Combined	O
expression	O
of	O
c-Jun	B-protein
and	O
p65	B-protein
induced	O
vigorous	O
transcription	O
of	O
IL-2	B-DNA
promoter-	I-DNA
and	I-DNA
CD28RE-driven	I-DNA
reporter	I-DNA
constructs	I-DNA
in	O
both	O
LFA-3-	B-cell_type
and	I-cell_type
B7-1-costimulated	I-cell_type
Jurkat	I-cell_type
cells	I-cell_type
.	O

Mutating	O
the	O
CD28RE	B-DNA
but	O
not	O
the	O
upstream	B-DNA
nuclear	I-DNA
factor	I-DNA
kappabeta-binding	I-DNA
site	I-DNA
in	O
the	O
IL-2	B-DNA
promoter	I-DNA
reduced	O
B7-1	B-protein
-driven	O
transcription	O
>	O
90	O
%	O
.	O

The	O
results	O
implicates	O
a	O
major	O
role	O
of	O
the	O
CD28RE	B-DNA
in	O
the	O
integration	O
of	O
p65	B-protein
/	O
c-Jun	B-protein
-mediated	O
transcription	O
within	O
the	O
IL-2	B-DNA
promoter	I-DNA
.	O

We	O
suggest	O
that	O
the	O
transition	O
from	O
an	O
autocrine	O
LFA-3	B-protein
-driven	O
immune	O
response	O
to	O
a	O
B7	O
--	O
induced	O
paracrine	O
immune	O
response	O
involves	O
the	O
activation	O
of	O
c-Jun	B-protein
and	O
p65	B-protein
,	O
which	O
target	O
the	O
CD28RE	B-DNA
region	O
of	O
the	O
IL-2	B-DNA
promoter	I-DNA
.	O

-DOCSTART-	O

Defects	O
in	O
actin-cap	O
formation	O
in	O
Vav-deficient	O
mice	O
implicate	O
an	O
actin	O
requirement	O
for	O
lymphocyte	O
signal	O
transduction	O
.	O

BACKGROUND	O
:	O
Antigen-receptor	O
interactions	O
on	O
lymphocytes	B-cell_type
result	O
in	O
local	O
clustering	O
of	O
actin	B-protein
,	O
receptors	O
and	O
signaling	O
molecules	O
into	O
an	O
asymmetric	O
membrane	O
structure	O
termed	O
a	O
cap	O
.	O

Although	O
actin	O
polymerization	O
is	O
known	O
to	O
be	O
required	O
,	O
the	O
mechanisms	O
underlying	O
cap	O
formation	O
are	O
unclear	O
.	O

We	O
have	O
studied	O
the	O
events	O
underlying	O
cap	O
formation	O
using	O
mice	O
bearing	O
a	O
null	O
mutation	O
in	O
vav	B-DNA
(	O
vav	B-DNA
-/-	O
)	O
,	O
a	O
gene	O
that	O
encodes	O
a	O
guanine-nucleotide	B-protein
exchange	I-protein
factor	I-protein
for	O
the	O
GTPase	B-protein
Rac	I-protein
.	O

RESULTS	O
:	O
Lymphocytes	B-cell_type
from	O
vav	B-DNA
-/-	O
mice	O
failed	O
to	O
form	O
T-cell	O
receptor	O
caps	O
following	O
activation	O
and	O
had	O
a	O
defective	O
actin	O
cytoskeleton	O
.	O

The	O
vav-/-	B-cell_type
T	I-cell_type
cells	I-cell_type
were	O
deficient	O
in	O
interleukin-2	O
(	O
IL-2	O
)	O
production	O
and	O
proliferation	O
,	O
and	O
the	O
peak	O
of	O
Ca2+	O
mobilization	O
was	O
reduced	O
although	O
of	O
normal	O
duration	O
.	O

Activation	O
of	O
Jun	B-protein
N-terminal	I-protein
kinase	I-protein
or	O
stress-activated	B-protein
kinase	I-protein
(	O
JNK	B-protein
or	O
SAPK	B-protein
)	O
and	O
mitogen-activated	B-protein
protein	I-protein
kinase	I-protein
(	O
MAPK	B-protein
)	O
and	O
the	O
induction	O
of	O
the	O
transcription	B-protein
factor	I-protein
NF-ATc1	B-DNA
and	I-DNA
egr-1	I-DNA
genes	I-DNA
was	O
normal	O
.	O

Despite	O
the	O
reduced	O
Ca2+	O
mobilization	O
,	O
translocation	O
of	O
cytoplasmic	B-protein
NF-ATc	I-protein
to	O
the	O
nucleus	O
was	O
normal	O
,	O
reflecting	O
that	O
the	O
lower	O
levels	O
of	O
Ca2+	O
in	O
vav-/-	B-cell_type
cells	I-cell_type
were	O
still	O
sufficient	O
to	O
activate	O
calcineurin	B-protein
.	O

Treatment	O
of	O
lymphocytes	B-cell_type
with	O
cytochalasin	O
D	O
,	O
which	O
blocks	O
actin	O
polymerization	O
,	O
inhibited	O
cap	O
formation	O
and	O
produced	O
defects	O
in	O
signaling	O
and	O
IL-2	O
transcriptional	O
induction	O
in	O
response	O
to	O
antigen-receptor	O
signaling	O
that	O
were	O
nearly	O
identical	O
to	O
those	O
seen	O
in	O
vav-/-	B-cell_type
cells	I-cell_type
.	O

In	O
transfection	O
studies	O
,	O
either	O
constitutively	O
active	O
Vav	B-protein
or	O
Rac	B-protein
could	O
complement	O
constitutively	O
active	O
calcineurin	B-protein
to	O
activate	O
NF-AT-dependent	O
transcription	O
.	O

CONCLUSIONS	O
:	O
These	O
results	O
indicate	O
that	O
Vav	B-protein
is	O
required	O
for	O
cap	O
formation	O
in	O
lymphocytes	B-cell_type
.	O

Furthermore	O
,	O
the	O
correlation	O
between	O
cap	O
formation	O
,	O
IL-2	O
production	O
and	O
proliferation	O
supports	O
the	O
hypothesis	O
that	O
an	O
actin-dependent	O
pathway	O
is	O
a	O
source	O
of	O
specialized	O
growth	O
regulatory	O
signals	O
.	O

-DOCSTART-	O

CD14	B-protein
-dependent	O
activation	O
of	O
human	B-cell_type
endothelial	I-cell_type
cells	I-cell_type
by	O
Bacteroides	O
fragilis	O
outer	O
membrane	O
.	O

We	O
studied	O
the	O
capacity	O
of	O
isolated	O
Bacteriodes	O
fragilis	O
outer	O
membrane	O
,	O
B.	O
fragilis	O
NCTC9343	O
lipopolysaccharide	O
(	O
LPS	O
;	O
endotoxin	O
)	O
,	O
and	O
B.	O
fragilis	O
NCTC9343	O
capsular	O
polysaccharides	O
to	O
activate	O
human	O
umbilical	O
vein	O
endothelial	O
cell	O
(	O
HUVEC	O
)	O
monolayers	O
.	O

To	O
assess	O
HUVEC	O
activation	O
,	O
E-selectin	B-protein
expression	O
was	O
measured	O
by	O
enzyme-linked	O
immunosorbent	O
assay	O
(	O
ELISA	O
)	O
,	O
Northern	O
blot	O
analysis	O
for	O
E-selectin-specific	B-RNA
mRNA	I-RNA
,	O
and	O
electrophoretic	O
gel	O
mobility	O
shift	O
assay	O
(	O
EMSA	O
)	O
for	O
NF-kappa	B-protein
B	I-protein
,	O
a	O
transcription	B-protein
factor	I-protein
necessary	O
for	O
E-selectin	B-DNA
gene	I-DNA
activation	O
.	O

Exposure	O
of	O
HUVECs	B-cell_type
to	O
B.	O
fragilis	O
outer	O
membrane	O
fractions	O
,	O
separated	O
from	O
other	O
components	O
of	O
the	O
B.	O
fragilis	O
cell	O
wall	O
by	O
isopycnic	O
,	O
sucrose	O
gradient	O
centrifugation	O
,	O
significantly	O
increased	O
surface	O
expression	O
of	O
E-selectin	B-protein
and	O
induced	O
functional	O
endothelial	B-cell_type
cell	I-cell_type
-dependent	O
leukocyte	O
adhesion	O
.	O

B.	O
fragilis	O
outer	O
membranes	O
induced	O
translocation	O
of	O
NF-kappa	B-protein
B	I-protein
to	O
HUVEC	O
nuclei	O
and	O
accumulation	O
of	O
E-selectin	B-RNA
mRNA	I-RNA
in	O
HUVEC	O
cytoplasm	O
.	O

E-selectin	O
expression	O
induced	O
by	O
B.	O
fragilis	O
outer	O
membranes	O
was	O
not	O
blocked	O
by	O
polymixin	O
B	O
.	O

In	O
contrast	O
,	O
E-selectin	O
expression	O
induced	O
by	O
outer	O
membrane	O
fractions	O
purified	O
from	O
E.	O
coli	O
was	O
competitively	O
inhibited	O
by	O
polymixin	O
B	O
.	O

Neither	O
purified	O
B.	O
fragilis	O
LPS	O
,	O
a	O
prominent	O
constituent	O
of	O
the	O
outer	O
membrane	O
,	O
nor	O
purified	O
B.	O
fragilis	O
capsular	O
polysaccharides	O
induced	O
HUVEC	O
activation	O
.	O

Two	O
different	O
monoclonal	B-protein
antibodies	I-protein
directed	O
against	O
human	B-protein
CD14	I-protein
completely	O
inhibited	O
B.	O
fragilis	O
outer	O
membrane-induced	O
NF-kappa	B-protein
B	I-protein
activation	O
,	O
E-selectin	O
transcription	O
,	O
and	O
E-selectin	O
surface	O
expression	O
.	O

We	O
conclude	O
that	O
the	O
outer	O
membrane	O
component	O
of	O
the	O
B.	O
fragilis	O
cell	O
wall	O
contains	O
a	O
proinflammatory	O
factor	O
(	O
s	O
)	O
,	O
that	O
is	O
not	O
LPS	O
,	O
which	O
induces	O
human	B-cell_type
endothelial	I-cell_type
cell	I-cell_type
activation	O
by	O
a	O
soluble	O
CD14	B-protein
-dependent	O
mechanism	O
.	O

-DOCSTART-	O

NF-kappaB	B-protein
protects	O
HIV-1-infected	O
myeloid	B-cell_type
cells	I-cell_type
from	O
apoptosis	O
.	O

HIV-1	O
infection	O
of	O
primary	B-cell_type
monocytic	I-cell_type
cells	I-cell_type
and	O
myeloid	B-cell_line
cell	I-cell_line
lines	I-cell_line
results	O
in	O
sustained	O
NF-kappaB	O
activation	O
.	O

Recently	O
,	O
NF-kappaB	O
induction	O
has	O
been	O
shown	O
to	O
play	O
a	O
role	O
in	O
protecting	O
cells	O
from	O
programmed	O
cell	O
death	O
.	O

In	O
the	O
present	O
study	O
,	O
we	O
sought	O
to	O
investigate	O
whether	O
constitutive	O
NF-kappaB	B-protein
activity	O
in	O
chronically	O
HIV-1-infected	O
promonocytic	B-cell_line
U937	I-cell_line
(	I-cell_line
U9-IIIB	I-cell_line
)	I-cell_line
and	I-cell_line
myeloblastic	I-cell_line
PLB-985	I-cell_line
(	I-cell_line
PLB-IIIB	I-cell_line
)	I-cell_line
cells	I-cell_line
affects	O
apoptotic	O
signaling	O
.	O

TNFalpha	B-protein
and	O
cycloheximide	O
caused	O
infected	O
cells	O
to	O
undergo	O
apoptosis	O
more	O
rapidly	O
than	O
parental	B-cell_line
U937	I-cell_line
and	O
PLB-985	B-cell_line
cells	I-cell_line
.	O

Inhibition	O
of	O
TNFalpha	B-protein
-induced	O
NF-kappaB	O
activation	O
using	O
the	O
antioxidant	O
N-acetylcysteine	O
(	O
NAC	O
)	O
resulted	O
in	O
increased	O
apoptosis	O
in	O
both	O
U937	B-cell_line
and	I-cell_line
U9-IIIB	I-cell_line
cells	I-cell_line
,	O
while	O
preactivation	O
of	O
NF-kappaB	B-protein
with	O
the	O
non-apoptotic	B-protein
inducer	I-protein
IL-1beta	I-protein
caused	O
a	O
relative	O
decrease	O
in	O
apoptosis	O
.	O

Inhibition	O
of	O
constitutive	O
NF-kappaB	B-protein
activity	O
in	O
U9-IIIB	B-cell_line
and	I-cell_line
PLB-IIIB	I-cell_line
cells	I-cell_line
also	O
induced	O
apoptosis	O
,	O
suggesting	O
that	O
NF-kappaB	B-cell_type
protects	I-cell_type
cells	I-cell_type
from	O
a	O
persistent	O
apoptotic	O
signal	O
.	O

TNFalpha	B-protein
plus	O
NAC	O
treatment	O
resulted	O
in	O
a	O
marked	O
decrease	O
in	O
Bcl-2	B-protein
protein	I-protein
levels	O
in	O
HIV-1-infected	B-cell_type
cells	I-cell_type
,	O
coupled	O
with	O
an	O
increase	O
in	O
Bax	B-protein
protein	I-protein
compared	O
to	O
uninfected	O
cells	O
,	O
suggesting	O
that	O
the	O
difference	O
in	O
susceptibility	O
to	O
TNFalpha	B-protein
-induced	O
apoptosis	O
may	O
relate	O
to	O
the	O
differences	O
in	O
relative	O
levels	O
of	O
Bcl-2	B-protein
and	O
Bax	B-protein
.	O

The	O
protective	O
role	O
of	O
NF-kappaB	B-protein
in	O
blocking	O
TNFalpha-	O
and	O
HIV-1-induced	O
apoptosis	O
was	O
supported	O
by	O
studies	O
in	O
Jurkat	B-cell_line
T	I-cell_line
cells	I-cell_line
engineered	O
to	O
express	O
IkappaB	B-protein
alpha	I-protein
repressor	I-protein
mutants	I-protein
(	O
TD-IkappaB	B-protein
)	O
under	O
the	O
control	O
of	O
a	O
tetracycline-responsive	B-DNA
promoter	I-DNA
.	O

Cells	O
underwent	O
apoptosis	O
in	O
response	O
to	O
TNFalpha	B-protein
only	O
when	O
NF-kappaB	O
activation	O
was	O
inhibited	O
by	O
TD-IkappaB	B-protein
expression	O
.	O

As	O
was	O
observed	O
for	O
the	O
U9-IIIB	B-cell_line
cells	I-cell_line
,	O
TNFalpha	B-protein
treatment	O
also	O
induced	O
a	O
marked	O
decrease	O
in	O
Bcl-2	B-protein
protein	I-protein
levels	O
in	O
TD-IkappaB	B-cell_type
expressing	I-cell_type
cells	I-cell_type
.	O

These	O
experiments	O
demonstrate	O
that	O
apoptotic	O
signaling	O
is	O
perturbed	O
in	O
HIV-1-infected	O
U9-IIIB	B-cell_line
cells	I-cell_line
and	O
indicate	O
that	O
NF-kappaB	O
activation	O
may	O
play	O
an	O
additional	O
protective	O
role	O
against	O
HIV-1-induced	O
apoptosis	O
in	O
myeloid	B-cell_type
cells	I-cell_type
.	O

-DOCSTART-	O

Human	B-cell_type
normal	I-cell_type
peripheral	I-cell_type
blood	I-cell_type
B-lymphocytes	I-cell_type
are	O
deficient	O
in	O
DNA-dependent	O
protein	O
kinase	O
activity	O
due	O
to	O
the	O
expression	O
of	O
a	O
variant	O
form	O
of	O
the	O
Ku86	B-protein
protein	I-protein
.	O

The	O
heterodimeric	O
Ku	B-protein
protein	I-protein
,	O
which	O
comprises	O
a	O
86	B-protein
kDa	I-protein
(	I-protein
Ku86	I-protein
)	I-protein
amd	O
a	O
70	B-protein
kDa	I-protein
(	I-protein
Ku70	I-protein
)	I-protein
subunits	I-protein
,	O
is	O
an	O
abundant	O
nuclear	B-protein
DNA-binding	I-protein
protein	I-protein
which	O
binds	O
in	O
vitro	O
to	O
DNA	B-DNA
termini	I-DNA
without	O
sequence	O
specificity	O
.	O

Ku	B-protein
is	O
the	O
DNA-targeting	B-protein
component	I-protein
of	O
the	O
large	O
catalytic	B-protein
sub-unit	I-protein
of	O
the	O
DNA-dependent	B-protein
protein	I-protein
kinase	I-protein
complex	I-protein
(	O
DNA-PK	B-protein
[	I-protein
CS	I-protein
]	I-protein
)	O
,	O
that	O
plays	O
a	O
critical	O
role	O
in	O
mammalian	O
double-strand	O
break	O
repair	O
and	O
lymphoid	O
V	O
(	O
D	O
)	O
J	O
recombination	O
.	O

By	O
using	O
electrophoretic	O
mobility	O
shift	O
assays	O
,	O
we	O
demonstrated	O
that	O
in	O
addition	O
to	O
the	O
major	B-protein
Ku	I-protein
x	I-protein
DNA	I-protein
complex	I-protein
usually	O
detected	O
in	O
cell	O
line	O
extracts	O
,	O
a	O
second	O
complex	O
with	O
faster	O
electrophoretic	O
mobility	O
was	O
observed	O
in	O
normal	O
peripheral	B-cell_type
blood	I-cell_type
lymphocytes	I-cell_type
(	O
PBL	B-cell_type
)	O
extracts	O
.	O

The	O
presence	O
of	O
this	O
faster	B-protein
migrating	I-protein
complex	I-protein
was	O
restricted	O
to	O
B	B-cell_type
cells	I-cell_type
among	O
the	O
circulating	B-cell_type
lymphocyte	I-cell_type
population	I-cell_type
.	O

Western	O
blot	O
analysis	O
revealed	O
that	O
B	B-cell_type
cells	I-cell_type
express	O
a	O
variant	O
form	O
of	O
the	O
Ku86	B-protein
protein	I-protein
with	O
an	O
apparent	O
molecular	O
weight	O
of	O
69	O
kDa	O
,	O
and	O
not	O
the	O
86	B-protein
kDa-	I-protein
full-length	I-protein
protein	I-protein
.	O

Although	O
the	O
heterodimer	B-protein
Ku70/variant-Ku86	I-protein
binds	O
to	O
DNA-ends	B-DNA
,	O
this	O
altered	O
form	O
of	O
the	O
Ku	B-protein
heterodimer	I-protein
has	O
a	O
decreased	O
ability	O
to	O
recruit	O
the	O
catalytic	B-protein
component	I-protein
of	O
the	O
complex	O
,	O
DNA-PK	B-protein
(	I-protein
CS	I-protein
)	I-protein
,	O
which	O
contributes	O
to	O
an	O
absence	O
of	O
detectable	O
DNA-PK	O
activity	O
in	O
B	B-cell_type
cells	I-cell_type
.	O

These	O
data	O
provide	O
a	O
molecular	O
basis	O
for	O
the	O
increased	O
sensitivity	O
of	O
B	B-cell_type
cells	I-cell_type
to	O
ionizing	O
radiation	O
and	O
identify	O
a	O
new	O
mechanism	O
of	O
regulation	O
of	O
DNA-PK	O
activity	O
that	O
operates	O
in	O
vivo	O
.	O

-DOCSTART-	O

Co-stimulation	O
of	O
human	B-cell_type
peripheral	I-cell_type
blood	I-cell_type
mononuclear	I-cell_type
cells	I-cell_type
with	O
IL-2	B-protein
and	O
anti-CD3	B-protein
monoclonal	I-protein
antibodies	I-protein
induces	O
phosphorylation	O
of	O
CREB	B-protein
.	O

Phosphorylation	O
of	O
the	O
cAMP-response	B-protein
element	I-protein
binding	I-protein
protein	I-protein
CREB	I-protein
within	O
1	O
h	O
of	O
CD2	B-protein
but	O
not	O
CD3	B-protein
cross-linking	O
of	O
human	B-cell_type
PBMC	I-cell_type
was	O
recently	O
demonstrated	O
.	O

The	O
absence	O
of	O
P-CREB	B-protein
following	O
CD3	O
cross-linking	O
was	O
unexpected	O
,	O
as	O
other	O
laboratories	O
reported	O
increased	O
phosphorylation	O
of	O
CREB	B-protein
following	O
CD3	O
cross-linking	O
of	O
the	O
Jurkat	B-cell_line
lymphocyte	I-cell_line
cell	I-cell_line
line	I-cell_line
.	O

Due	O
to	O
Jurkat	B-cell_line
T-cells	I-cell_line
being	O
IL-2-independent	O
,	O
it	O
was	O
postulated	O
that	O
IL-2	B-protein
might	O
provide	O
a	O
necessary	O
co-stimulus	O
for	O
phosphorylation	O
of	O
CREB	B-protein
in	O
primary	B-cell_type
lymphocytes	I-cell_type
.	O

Therefore	O
,	O
P-CREB	B-protein
was	O
evaluated	O
following	O
co-stimulation	O
of	O
human	B-cell_type
PBMC	I-cell_type
through	O
the	O
IL-2	B-protein
and	I-protein
CD2	I-protein
or	I-protein
CD3	I-protein
receptors	I-protein
.	O

IL-2	B-protein
did	O
not	O
further	O
augment	O
phosphorylation	O
of	O
CREB	B-protein
following	O
CD2	O
cross-linking	O
.	O

However	O
,	O
while	O
neither	O
IL-2	B-protein
nor	O
CD3	O
cross-linking	O
alone	O
induced	O
P-CREB	B-protein
,	O
a	O
4.5-fold	O
increase	O
in	O
phosphorylation	O
of	O
CREB	B-protein
within	O
1	O
h	O
of	O
IL-2/CD3	O
co-stimulation	O
was	O
observed	O
.	O

Phosphorylation	O
was	O
not	O
associated	O
with	O
the	O
induction	O
of	O
cAMP	O
,	O
and	O
inhibition	O
of	O
PKA	O
signaling	O
had	O
no	O
effect	O
on	O
P-CREB	B-protein
.	O

Consistent	O
with	O
signal	O
transduction	O
through	O
p56lck	B-protein
or	O
p59fyn	B-protein
,	O
inhibition	O
of	O
PTK	O
signaling	O
reduced	O
phosphorylation	O
50	O
%	O
.	O

Interestingly	O
,	O
inhibiting	O
PKC	O
signaling	O
with	O
calphostin	O
C	O
further	O
increased	O
P-CREB	O
levels	O
3-fold	O
over	O
that	O
observed	O
in	O
IL-2/CD3	B-cell_type
co-stimulated	I-cell_type
cells	I-cell_type
not	O
pretreated	O
with	O
a	O
PKC	O
inhibitor	O
.	O

In	O
contrast	O
to	O
previous	O
studies	O
performed	O
in	O
the	O
absence	O
of	O
exogenous	B-protein
IL-2	I-protein
,	O
no	O
increase	O
in	O
binding	O
of	O
CREB	B-protein
to	O
a	O
32P-labeled	O
oligonucleotide	O
probe	O
was	O
observed	O
by	O
electrophoretic	O
mobility	O
shift	O
assay	O
.	O

These	O
data	O
suggest	O
that	O
the	O
IL-2	B-protein
and	O
CD3	B-protein
signaling	O
pathways	O
provide	O
a	O
necessary	O
and	O
co-operative	O
stimulus	O
promoting	O
phosphorylation	O
of	O
CREB	B-protein
following	O
receptor	O
cross-linking	O
.	O

-DOCSTART-	O

HIV-1	O
infection	O
induces	O
a	O
selective	O
reduction	O
in	O
STAT5	O
protein	O
expression	O
.	O

HIV-1	O
infection	O
is	O
accompanied	O
by	O
qualitative	O
and	O
quantitative	O
defects	O
in	O
CD4+	B-cell_type
T	I-cell_type
lymphocytes	I-cell_type
.	O

Loss	O
of	O
immune	O
function	O
in	O
HIV	O
patients	O
is	O
usually	O
associated	O
with	O
a	O
profound	O
dysregulation	O
of	O
cytokine	B-protein
production	O
.	O

To	O
investigate	O
whether	O
cytokine	B-protein
signaling	O
defects	O
occur	O
during	O
HIV	O
infection	O
,	O
PHA	B-cell_type
blasts	I-cell_type
from	O
healthy	O
human	O
donors	O
were	O
infected	O
with	O
two	O
strains	O
of	O
HIV-1	O
and	O
screened	O
for	O
the	O
expression	O
of	O
STAT	B-protein
proteins	I-protein
used	O
in	O
cytokine	B-protein
signaling	O
.	O

A	O
selective	O
decrease	O
in	O
STAT5B	B-protein
was	O
seen	O
8	O
days	O
after	O
infection	O
with	O
the	O
BZ167	O
dual-tropic	O
HIV	O
isolate	O
,	O
but	O
not	O
with	O
the	O
Ba-L	O
,	O
M-tropic	O
strain	O
.	O

Based	O
on	O
these	O
findings	O
,	O
purified	B-cell_type
T	I-cell_type
cells	I-cell_type
from	O
HIV-infected	O
patients	O
in	O
different	O
stages	O
of	O
disease	O
were	O
also	O
tested	O
for	O
STAT	O
expression	O
;	O
decreases	O
in	O
STAT5A	B-protein
,	O
STAT5B	B-protein
,	O
and	O
STAT1alpha	B-protein
were	O
observed	O
in	O
all	O
patients	O
.	O

The	O
reduction	O
in	O
STATs	B-protein
seen	O
in	O
vivo	O
and	O
in	O
vitro	O
after	O
HIV	O
infection	O
may	O
contribute	O
to	O
the	O
loss	O
of	O
T	B-cell_type
cell	I-cell_type
function	O
in	O
HIV	O
disease	O
.	O

-DOCSTART-	O

Transcription	B-protein
factors	I-protein
that	O
regulate	O
monocyte	B-cell_type
/	O
macrophage	B-cell_type
differentiation	O
.	O

Although	O
all	O
the	O
cells	O
in	O
an	O
organism	O
contain	O
the	O
same	O
genetic	O
information	O
,	O
differences	O
in	O
the	O
cell	O
phenotype	O
arise	O
from	O
the	O
expression	O
of	O
lineage-specific	B-DNA
genes	I-DNA
.	O

During	O
myelopoiesis	O
,	O
external	O
differentiating	O
signals	O
regulate	O
the	O
expression	O
of	O
a	O
set	O
of	O
transcription	B-protein
factors	I-protein
.	O

The	O
combined	O
action	O
of	O
these	O
transcription	B-protein
factors	I-protein
subsequently	O
determines	O
the	O
expression	O
of	O
myeloid-specific	B-DNA
genes	I-DNA
and	O
the	O
generation	O
of	O
monocytes	B-cell_type
and	O
macrophages	B-cell_type
.	O

In	O
particular	O
,	O
the	O
transcription	B-protein
factor	I-protein
PU.1	I-protein
has	O
a	O
critical	O
role	O
in	O
this	O
process	O
.	O

We	O
review	O
the	O
contribution	O
of	O
several	O
transcription	B-protein
factors	I-protein
to	O
the	O
control	O
of	O
macrophage	B-cell_type
development	O

-DOCSTART-	O

Transcription	B-protein
factor	I-protein
LKLF	I-protein
is	O
sufficient	O
to	O
program	O
T	O
cell	O
quiescence	O
via	O
a	O
c-Myc	O
--	O
dependent	O
pathway	O
.	O

T	B-cell_type
lymphocytes	I-cell_type
circulate	O
in	O
a	O
quiescent	O
state	O
until	O
they	O
encounter	O
cognate	B-protein
antigen	I-protein
bound	O
to	O
the	O
surface	O
of	O
an	O
antigen-presenting	B-cell_type
cell	I-cell_type
.	O

The	O
molecular	O
pathways	O
that	O
regulate	O
T	O
cell	O
quiescence	O
remain	O
largely	O
unknown	O
.	O

Here	O
we	O
show	O
that	O
forced	O
expression	O
of	O
the	O
lung	B-protein
Kruppel-like	I-protein
transcription	I-protein
factor	I-protein
(	O
LKLF	B-protein
)	O
in	O
Jurkat	B-cell_line
T	I-cell_line
cells	I-cell_line
is	O
sufficient	O
to	O
program	O
a	O
quiescent	O
phenotype	O
characterized	O
by	O
decreased	O
proliferation	O
,	O
reduced	O
cell	O
size	O
and	O
protein	O
synthesis	O
and	O
decreased	O
surface	O
expression	O
of	O
activation	O
markers	O
.	O

Conversely	O
,	O
LKLF-deficient	B-cell_type
peripheral	I-cell_type
T	I-cell_type
cells	I-cell_type
produced	O
by	O
gene	O
targeting	O
showed	O
increased	O
proliferation	O
,	O
increased	O
cell	O
size	O
and	O
enhanced	O
expression	O
of	O
surface	O
activation	O
markers	O
in	O
vivo	O
.	O

LKLF	B-protein
appeared	O
to	O
function	O
,	O
at	O
least	O
in	O
part	O
,	O
by	O
decreasing	O
expression	O
of	O
the	O
proto-oncogene	B-DNA
encoding	O
c-Myc	B-protein
.	O

Forced	O
expression	O
of	O
LKLF	B-protein
was	O
associated	O
with	O
markedly	O
decreased	O
c-Myc	B-protein
expression	O
.	O

In	O
addition	O
,	O
many	O
effects	O
of	O
LKLF	B-protein
expression	O
were	O
mimicked	O
by	O
expression	O
of	O
the	O
dominant-negative	B-protein
MadMyc	I-protein
protein	I-protein
and	O
rescued	O
by	O
overexpression	O
of	O
c-Myc	B-protein
.	O

Thus	O
,	O
LKLF	B-protein
is	O
both	O
necessary	O
and	O
sufficient	O
to	O
program	O
quiescence	O
in	O
T	B-cell_type
cells	I-cell_type
and	O
functions	O
,	O
in	O
part	O
,	O
by	O
negatively	O
regulating	O
a	O
c-Myc	O
--	O
dependent	O
pathway	O
.	O

-DOCSTART-	O

HTLV-1	B-protein
p12	I-protein
(	I-protein
I	I-protein
)	I-protein
protein	I-protein
enhances	O
STAT5	O
activation	O
and	O
decreases	O
the	O
interleukin-2	B-protein
requirement	O
for	O
proliferation	O
of	O
primary	B-cell_type
human	I-cell_type
peripheral	I-cell_type
blood	I-cell_type
mononuclear	I-cell_type
cells	I-cell_type
.	O

The	O
p12	B-protein
(	I-protein
I	I-protein
)	I-protein
protein	I-protein
,	O
encoded	O
by	O
the	O
pX	B-DNA
open	I-DNA
reading	I-DNA
frame	I-DNA
I	I-DNA
of	O
the	O
human	O
T-lymphotropic	O
virus	O
type	O
1	O
(	O
HTLV-1	O
)	O
,	O
is	O
a	O
hydrophobic	B-protein
protein	I-protein
that	O
localizes	O
to	O
the	O
endoplasmic	O
reticulum	O
and	O
the	O
Golgi	O
.	O

Although	O
p12	B-protein
(	I-protein
I	I-protein
)	I-protein
contains	O
4	O
minimal	O
proline-rich	O
,	O
src	B-protein
homology	I-protein
3-binding	I-protein
motifs	I-protein
(	O
PXXP	B-protein
)	O
,	O
a	O
characteristic	O
commonly	O
found	O
in	O
proteins	O
involved	O
in	O
signaling	O
pathways	O
,	O
it	O
has	O
not	O
been	O
known	O
whether	O
p12	B-protein
(	I-protein
I	I-protein
)	I-protein
has	O
a	O
role	O
in	O
modulating	O
intracellular	O
signaling	O
pathways	O
.	O

This	O
study	O
demonstrated	O
that	O
p12	B-protein
(	I-protein
I	I-protein
)	I-protein
binds	O
to	O
the	O
cytoplasmic	B-protein
domain	I-protein
of	O
the	O
interleukin-2	B-protein
receptor	I-protein
(	I-protein
IL-2R	I-protein
)	I-protein
beta	I-protein
chain	I-protein
that	O
is	O
involved	O
in	O
the	O
recruitment	O
of	O
the	O
Jak1	B-protein
and	I-protein
Jak3	I-protein
kinases	I-protein
.	O

As	O
a	O
result	O
of	O
this	O
interaction	O
,	O
p12	B-protein
(	I-protein
I	I-protein
)	I-protein
increases	O
signal	B-protein
transducers	I-protein
and	I-protein
activators	I-protein
of	I-protein
transcription	I-protein
5	I-protein
(	O
STAT5	B-protein
)	O
DNA	O
binding	O
and	O
transcriptional	O
activity	O
and	O
this	O
effect	O
depends	O
on	O
the	O
presence	O
of	O
both	O
IL-2R	B-protein
beta	I-protein
and	I-protein
gamma	I-protein
(	I-protein
c	I-protein
)	I-protein
chains	I-protein
and	O
Jak3	B-protein
.	O

Transduction	O
of	O
primary	B-cell_line
human	I-cell_line
peripheral	I-cell_line
blood	I-cell_line
mononuclear	I-cell_line
cells	I-cell_line
(	O
PBMCs	B-cell_type
)	O
with	O
a	O
human	B-DNA
immunodeficiency	I-DNA
virus	I-DNA
type	I-DNA
1-based	I-DNA
retroviral	I-DNA
vector	I-DNA
expressing	O
p12	B-protein
(	I-protein
I	I-protein
)	I-protein
also	O
resulted	O
in	O
increased	O
STAT5	B-protein
phosphorylation	O
and	O
DNA	O
binding	O
.	O

However	O
,	O
p12	B-protein
(	I-protein
I	I-protein
)	I-protein
could	O
increase	O
proliferation	O
of	O
human	B-cell_type
PBMCs	I-cell_type
only	O
after	O
stimulation	O
of	O
T-cell	B-protein
receptors	I-protein
by	O
treatment	O
of	O
cells	O
with	O
low	O
concentrations	O
of	O
alphaCD3	B-protein
and	I-protein
alphaCD28	I-protein
antibodies	I-protein
.	O

In	O
addition	O
,	O
the	O
proliferative	O
advantage	O
of	O
p12	B-cell_type
(	I-cell_type
I	I-cell_type
)	I-cell_type
-transduced	I-cell_type
PBMCs	I-cell_type
was	O
evident	O
mainly	O
at	O
low	O
concentrations	O
of	O
IL-2	B-protein
.	O

Together	O
,	O
these	O
data	O
indicate	O
that	O
p12	B-protein
(	I-protein
I	I-protein
)	I-protein
may	O
confer	O
a	O
proliferative	O
advantage	O
on	O
HTLV-1-infected	B-cell_type
cells	I-cell_type
in	O
the	O
presence	O
of	O
suboptimal	O
antigen	O
stimulation	O
and	O
that	O
this	O
event	O
may	O
account	O
for	O
the	O
clonal	O
proliferation	O
of	O
infected	B-cell_type
T	I-cell_type
cells	I-cell_type
in	O
vivo	O
.	O

(	O
Blood.	O
2001	O
;	O
98	O
:	O
823-829	O
)	O

-DOCSTART-	O

Single	O
dose	O
intranasal	O
administration	O
of	O
retinal	B-protein
autoantigen	I-protein
generates	O
a	O
rapid	O
accumulation	O
and	O
cell	O
activation	O
in	O
draining	O
lymph	O
node	O
and	O
spleen	O
:	O
implications	O
for	O
tolerance	O
therapy	O
.	O

BACKGROUND/AIMS	O
:	O
A	O
single	O
intranasal	O
delivery	O
of	O
retinal	B-protein
autoantigen	I-protein
suppresses	O
effectively	O
experimental	O
autoimmune	O
uveoretinitis	O
(	O
EAU	O
)	O
.	O

To	O
further	O
unravel	O
underlying	O
mechanisms	O
the	O
authors	O
wished	O
to	O
determine	O
,	O
firstly	O
,	O
the	O
kinetics	O
of	O
antigen	O
delivery	O
and	O
,	O
secondly	O
,	O
the	O
early	O
cellular	O
responses	O
involved	O
in	O
the	O
initial	O
stages	O
of	O
nasal	O
mucosal	O
tolerance	O
induction	O
.	O

METHODS	O
:	O
Flow	O
cytometry	O
,	O
cell	O
proliferation	O
assays	O
,	O
and	O
microscopy	O
were	O
used	O
to	O
track	O
antigen	B-protein
following	O
a	O
single	O
,	O
intranasal	O
dose	O
of	O
Alexa-488	B-protein
labelled	I-protein
retinal	I-protein
antigen	I-protein
.	O

RESULTS	O
:	O
A	O
rapid	O
accumulation	O
of	O
antigen	B-protein
within	O
both	O
superficial	O
cervical	O
lymph	O
nodes	O
(	O
SCLN	O
)	O
and	O
spleen	O
was	O
observed	O
after	O
30	O
minutes	O
.	O

Significant	O
proliferative	O
responses	O
to	O
IRBP	O
were	O
elicited	O
by	O
48	O
hours	O
indicating	O
that	O
systemic	O
priming	O
of	O
naive	B-cell_type
T	I-cell_type
cells	I-cell_type
to	O
retinal	B-protein
antigen	I-protein
had	O
occurred	O
.	O

Cell	O
activation	O
was	O
further	O
confirmed	O
by	O
immunoprecipitation	O
studies	O
,	O
which	O
demonstrated	O
phosphorylation	O
of	O
STAT4	B-protein
but	O
not	O
STAT6	B-protein
in	O
both	O
lymph	O
nodes	O
and	O
spleen	O
.	O

However	O
,	O
at	O
24	O
hours	O
,	O
STAT4	O
heterodimerisation	O
with	O
STAT	B-protein
3	I-protein
was	O
only	O
observed	O
in	O
spleen	O
.	O

CONCLUSIONS	O
:	O
The	O
results	O
provide	O
novel	O
evidence	O
that	O
following	O
a	O
single	O
intranasal	O
application	O
rapid	O
transfer	O
of	O
antigen	B-protein
occurs	O
.	O

Resulting	O
T	O
cell	O
proliferation	O
develops	O
consequent	O
to	O
differential	O
cell	O
signalling	O
in	O
SCLN	O
and	O
spleen	O
.	O

Further	O
understanding	O
of	O
these	O
underlying	O
cellular	O
mechanisms	O
,	O
in	O
particular	O
as	O
is	O
inferred	O
by	O
the	O
results	O
the	O
contribution	O
of	O
local	O
versus	O
systemic	O
tolerance	O
induction	O
,	O
may	O
assist	O
in	O
strategies	O
to	O
clinically	O
apply	O
mucosal	O
tolerance	O
therapy	O
successfully	O
.	O

-DOCSTART-	O

Regulation	O
of	O
interleukin	B-protein
(	I-protein
IL	I-protein
)	I-protein
-18	I-protein
receptor	I-protein
alpha	I-protein
chain	I-protein
expression	O
on	O
CD4	B-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
T	I-cell_type
cells	I-cell_type
during	O
T	O
helper	O
(	O
Th	O
)	O
1/Th2	O
differentiation	O
.	O

Critical	O
downregulatory	O
role	O
of	O
IL-4	B-protein
.	O

Interleukin	B-protein
(	I-protein
IL	I-protein
)	I-protein
-18	I-protein
has	O
been	O
well	O
characterized	O
as	O
a	O
costimulatory	B-protein
factor	I-protein
for	O
the	O
induction	O
of	O
IL-12-mediated	B-protein
interferon	I-protein
(	I-protein
IFN	I-protein
)	I-protein
-gamma	I-protein
production	O
by	O
T	B-cell_type
helper	I-cell_type
(	I-cell_type
Th	I-cell_type
)	I-cell_type
1	I-cell_type
cells	I-cell_type
,	O
but	O
also	O
can	O
induce	O
IL-4	O
production	O
and	O
thus	O
facilitate	O
the	O
differentiation	O
of	O
Th2	B-cell_type
cells	I-cell_type
.	O

To	O
determine	O
the	O
mechanisms	O
by	O
which	O
IL-18	B-protein
might	O
regulate	O
these	O
diametrically	O
distinct	O
immune	O
responses	O
,	O
we	O
have	O
analyzed	O
the	O
role	O
of	O
cytokines	B-protein
in	O
the	O
regulation	O
of	O
IL-18	B-protein
receptor	I-protein
alpha	I-protein
chain	I-protein
(	O
IL-18Ralpha	B-protein
)	O
expression	O
.	O

The	O
majority	O
of	O
peripheral	B-cell_type
CD4	I-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
T	I-cell_type
cells	I-cell_type
constitutively	O
expressed	O
the	O
IL-18Ralpha	B-protein
.	O

Upon	O
antigen	O
stimulation	O
in	O
the	O
presence	O
of	O
IL-12	B-protein
,	O
marked	O
enhancement	O
of	O
IL-18Ralpha	B-protein
expression	O
was	O
observed	O
.	O

IL-12	B-protein
-mediated	O
upregulation	O
of	O
IL-18Ralpha	B-protein
required	O
IFN-gamma	B-protein
.	O

Activated	B-cell_type
CD4	I-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
T	I-cell_type
cells	I-cell_type
that	O
expressed	O
low	O
levels	O
of	O
IL-18Ralpha	B-protein
could	O
produce	O
IFN-gamma	B-protein
when	O
stimulated	O
with	O
the	O
combination	O
of	O
IL-12	B-protein
and	O
IL-18	B-protein
,	O
while	O
CD4	B-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
cells	I-cell_type
which	O
expressed	O
high	O
levels	O
of	O
IL-18Ralpha	B-protein
could	O
respond	O
to	O
IL-18	B-protein
alone	O
.	O

In	O
contrast	O
,	O
T	O
cell	O
stimulation	O
in	O
the	O
presence	O
of	O
IL-4	B-protein
resulted	O
in	O
a	O
downregulation	O
of	O
IL-18Ralpha	B-protein
expression	O
.	O

Both	O
IL-4	B-cell_type
(	I-cell_type
-/	I-cell_type
)	I-cell_type
-	I-cell_type
and	I-cell_type
signal	I-cell_type
transducer	I-cell_type
and	I-cell_type
activator	I-cell_type
of	I-cell_type
transcription	I-cell_type
(	I-cell_type
Stat	I-cell_type
)	I-cell_type
6	I-cell_type
(	I-cell_type
-/	I-cell_type
)	I-cell_type
-	I-cell_type
T	I-cell_type
cells	I-cell_type
expressed	O
higher	O
levels	O
of	O
IL-18Ralpha	B-protein
after	O
TCR	O
stimulation	O
.	O

Furthermore	O
,	O
activated	B-cell_type
T	I-cell_type
cells	I-cell_type
from	O
Stat6	O
(	O
-/	O
)	O
-	O
mice	O
produced	O
more	O
IFN-gamma	B-protein
in	O
response	O
to	O
IL-18	B-protein
than	O
wild-type	O
controls	O
.	O

Thus	O
,	O
positive/negative	O
regulation	O
of	O
the	O
IL-18Ralpha	B-protein
by	O
the	O
major	O
inductive	O
cytokines	B-protein
(	O
IL-12	B-protein
and	O
IL-4	B-protein
)	O
determines	O
the	O
capacity	O
of	O
IL-18	B-protein
to	O
polarize	O
an	O
immune	O
response	O
.	O

-DOCSTART-	O

Glucocorticoid-regulated	B-protein
transcription	I-protein
factors	I-protein
.	O

Glucocorticoids	O
are	O
the	O
most	O
effective	O
antiinflammatory	O
drugs	O
used	O
in	O
the	O
treatment	O
of	O
asthma	O
.	O

They	O
act	O
by	O
binding	O
to	O
a	O
specific	O
receptor	O
(	O
GR	B-protein
)	O
that	O
,	O
upon	O
activation	O
,	O
translocates	O
to	O
the	O
nucleus	O
and	O
either	O
increases	O
(	O
transactivates	O
)	O
or	O
decreases	O
(	O
transrepresses	O
)	O
gene	O
expression	O
.	O

Inhibition	O
of	O
pro-inflammatory	B-protein
transcription	I-protein
factors	I-protein
such	O
as	O
activator	B-protein
protein	I-protein
(	I-protein
AP	I-protein
)	I-protein
-1	I-protein
,	O
signal	B-protein
transducers	I-protein
and	I-protein
activators	I-protein
of	I-protein
transcription	I-protein
(	O
STATs	B-protein
)	O
,	O
nuclear	B-protein
factor	I-protein
of	I-protein
activated	I-protein
T	I-protein
cells	I-protein
(	O
NFAT	B-protein
)	O
and	O
nuclear	B-protein
factor	I-protein
(	I-protein
NF	I-protein
)	I-protein
-kappa	I-protein
B	I-protein
is	O
thought	O
to	O
be	O
a	O
major	O
action	O
of	O
glucocorticoids	O
.	O

Acetylation	O
of	O
histones	B-protein
allows	O
unwinding	O
of	O
the	O
local	B-DNA
DNA	I-DNA
structure	I-DNA
and	O
enables	O
RNA	B-protein
polymerase	I-protein
II	I-protein
to	O
enhance	O
gene	O
transcription	O
.	O

Histone	O
acetylation	O
is	O
regulated	O
by	O
a	O
balance	O
between	O
the	O
activity	O
of	O
histone	B-protein
acetyltransferases	I-protein
(	O
HATs	B-protein
)	O
and	O
histone	B-protein
deacetylases	I-protein
(	O
HDACs	B-protein
)	O
.	O

GR	B-protein
acts	O
as	O
a	O
direct	O
inhibitor	O
of	O
NF-kappa	O
B-induced	O
HAT	O
activity	O
and	O
also	O
by	O
recruiting	O
HDAC2	B-protein
to	O
the	O
NF-kappa	B-protein
B/HAT	I-protein
complex	I-protein
.	O

A	O
sub-group	O
of	O
patients	O
with	O
glucocorticoid-insensitive	O
asthma	O
have	O
an	O
inability	O
to	O
induce	O
histone	O
acetylation	O
in	O
response	O
to	O
dexamethasone	O
suggesting	O
reduced	O
expression	O
of	O
a	O
GR-specific	B-protein
HAT	I-protein
.	O

This	O
suggests	O
that	O
pharmacological	O
manipulation	O
of	O
specific	O
histone	O
acetylation	O
status	O
is	O
a	O
potentially	O
useful	O
approach	O
for	O
the	O
treatment	O
of	O
inflammatory	O
diseases	O
.	O

Identification	O
of	O
the	O
precise	O
mechanism	O
by	O
which	O
activated	B-protein
GR	I-protein
recruits	O
HDAC2	B-protein
may	O
reveal	O
new	O
targets	O
for	O
the	O
development	O
of	O
drugs	O
that	O
may	O
dissociate	O
the	O
antiinflammatory	O
actions	O
of	O
glucocorticoids	O
from	O
their	O
side	O
effects	O
that	O
are	O
largely	O
due	O
to	O
gene	O
induction	O
.	O

Copyright	O
Academic	O
Press	O
.	O

-DOCSTART-	O

Differential	O
ultraviolet-B-induced	O
immunomodulation	O
in	O
XPA	O
,	O
XPC	O
,	O
and	O
CSB	O
DNA	O
repair-deficient	O
mice	O
.	O

Ultraviolet	O
B	O
irradiation	O
has	O
serious	O
consequences	O
for	O
cellular	O
immunity	O
and	O
can	O
suppress	O
the	O
rejection	O
of	O
skin	O
tumors	O
and	O
the	O
resistance	O
to	O
infectious	O
diseases	O
.	O

DNA	O
damage	O
plays	O
a	O
crucial	O
role	O
in	O
these	O
immunomodulatory	O
effects	O
of	O
ultraviolet	O
B	O
,	O
as	O
impaired	O
repair	O
of	O
ultraviolet-B-induced	O
DNA	O
damage	O
has	O
been	O
shown	O
to	O
cause	O
suppression	O
of	O
cellular	O
immunity	O
.	O

Ultraviolet-B-induced	O
DNA	O
damage	O
is	O
repaired	O
by	O
the	O
nucleotide	O
excision	O
repair	O
mechanism	O
very	O
efficiently	O
.	O

Nucleotide	O
excision	O
repair	O
comprises	O
two	O
subpathways	O
:	O
transcription-coupled	O
and	O
global	O
genome	O
repair	O
.	O

In	O
this	O
study	O
the	O
immunologic	O
consequences	O
of	O
specific	O
nucleotide	O
excision	O
repair	O
defects	O
in	O
three	O
mouse	O
models	O
,	O
XPA	O
,	O
XPC	O
,	O
and	O
CSB	O
mutant	O
mice	O
,	O
were	O
investigated	O
.	O

XPA	O
mice	O
carry	O
a	O
total	O
nucleotide	O
excision	O
repair	O
defect	O
,	O
whereas	O
XPC	O
and	O
CSB	O
mice	O
only	O
lack	O
global	O
genome	O
and	O
transcription-coupled	O
nucleotide	O
excision	O
repair	O
,	O
respectively	O
.	O

Our	O
data	O
demonstrate	O
that	O
cellular	O
immune	O
parameters	O
in	O
XPA	O
,	O
XPC	O
,	O
and	O
CSB	O
mice	O
are	O
normal	O
compared	O
with	O
their	O
wild-type	O
(	O
control	O
)	O
littermates	O
.	O

This	O
may	O
indicate	O
that	O
the	O
reported	O
altered	O
cellular	O
responses	O
in	O
xeroderma	O
pigmentosum	O
patients	O
are	O
not	O
constitutive	O
but	O
could	O
be	O
due	O
to	O
external	O
factors	O
,	O
such	O
as	O
ultraviolet	O
B	O
.	O

Upon	O
exposure	O
to	O
ultraviolet	O
B	O
,	O
only	O
XPA	O
mice	O
are	O
very	O
sensitive	O
to	O
ultraviolet-B-induced	O
inhibition	O
of	O
Th1-mediated	O
contact	O
hypersensitivity	O
responses	O
and	O
interferon-gamma	B-protein
production	O
in	O
skin	O
draining	O
lymph	O
nodes	O
.	O

Lipopolysaccharide-stimulated	O
tumor	B-protein
necrosis	I-protein
factor	I-protein
alpha	I-protein
and	O
interleukin-10	B-protein
production	O
are	O
significantly	O
augmented	O
in	O
both	O
XPA	O
and	O
CSB	O
mice	O
after	O
ultraviolet	O
B	O
exposure	O
.	O

Lymph	O
node	O
cell	O
numbers	O
were	O
increased	O
very	O
significantly	O
in	O
XPA	O
,	O
mildly	O
increased	O
in	O
CSB	O
,	O
and	O
not	O
in	O
XPC	O
mice	O
.	O

In	O
general	O
XPC	O
mice	O
do	O
not	O
exhibit	O
any	O
indication	O
of	O
enhanced	O
ultraviolet	O
B	O
susceptibility	O
with	O
regard	O
to	O
the	O
immune	O
parameters	O
analyzed	O
.	O

These	O
data	O
suggest	O
that	O
both	O
global	O
genome	O
repair	O
and	O
transcription-coupled	O
repair	O
are	O
needed	O
to	O
prevent	O
immunomodulation	O
by	O
ultraviolet	O
B	O
,	O
whereas	O
transcription-coupled	O
repair	O
is	O
the	O
major	O
DNA	O
repair	O
subpathway	O
of	O
nucleotide	O
excision	O
repair	O
that	O
prevents	O
the	O
acute	O
ultraviolet-B-induced	O
effects	O
such	O
as	O
erythema	O
.	O

-DOCSTART-	O

Biomechanical	O
strain	O
induces	O
class	O
a	O
scavenger	B-protein
receptor	I-protein
expression	O
in	O
human	B-cell_type
monocyte/macrophages	I-cell_type
and	O
THP-1	B-cell_type
cells	I-cell_type
:	O
a	O
potential	O
mechanism	O
of	O
increased	O
atherosclerosis	O
in	O
hypertension	O
.	O

BACKGROUND	O
:	O
Although	O
hypertension	O
is	O
an	O
important	O
risk	O
factor	O
for	O
the	O
development	O
of	O
atherosclerosis	O
,	O
the	O
mechanisms	O
for	O
this	O
interaction	O
are	O
incompletely	O
described	O
.	O

Previous	O
studies	O
have	O
suggested	O
that	O
biomechanical	O
strain	O
regulates	O
macrophage	O
phenotype	O
.	O

We	O
tested	O
the	O
hypothesis	O
that	O
biomechanical	O
strain	O
can	O
induce	O
expression	O
of	O
the	O
class	B-protein
A	I-protein
scavenger	I-protein
receptor	I-protein
(	O
SRA	B-protein
)	O
,	O
an	O
important	O
lipoprotein	B-protein
receptor	I-protein
in	O
atherogenesis	O
.	O

METHODS	O
AND	O
RESULTS	O
:	O
Human	B-cell_type
monocyte/macrophages	I-cell_type
or	O
THP-1	B-cell_type
cells	I-cell_type
were	O
cultured	O
in	O
a	O
device	O
that	O
imposes	O
uniform	O
biaxial	O
cyclic	O
1-Hz	O
strains	O
of	O
0	O
%	O
,	O
1	O
%	O
,	O
2	O
%	O
,	O
or	O
3	O
%	O
,	O
and	O
SRA	B-protein
expression	O
was	O
analyzed	O
.	O

Mechanical	O
strains	O
induced	O
SRA	B-RNA
mRNA	I-RNA
(	O
3.5+/-0.6-fold	O
at	O
3	O
%	O
strain	O
for	O
48	O
hours	O
,	O
P	O
<	O
0.01	O
)	O
and	O
SRA	B-protein
protein	I-protein
in	O
THP-1	B-cell_type
cells	I-cell_type
in	O
an	O
amplitude-dependent	O
manner	O
.	O

This	O
induction	O
was	O
accompanied	O
by	O
augmented	O
expression	O
of	O
the	O
class	B-protein
B	I-protein
scavenger	I-protein
receptor	I-protein
CD36	I-protein
(	O
2.8+/-0.3-fold	O
,	O
P	O
<	O
0.001	O
)	O
but	O
not	O
by	O
increased	O
peroxisome	B-protein
proliferator-activated	I-protein
receptor-gamma	I-protein
expression	O
.	O

To	O
evaluate	O
this	O
effect	O
in	O
vivo	O
,	O
apolipoprotein	O
E	O
(	O
-/-	O
)	O
mice	O
were	O
randomly	O
assigned	O
to	O
receive	O
standard	O
chow	O
,	O
a	O
high-cholesterol	O
diet	O
,	O
or	O
a	O
high-cholesterol	O
diet	O
with	O
hypertension	O
induced	O
by	O
angiotensin	O
II	O
infusion	O
for	O
8	O
weeks	O
.	O

Immunohistochemistry	O
revealed	O
that	O
among	O
macrophages	B-cell_type
in	O
atherosclerotic	O
lesions	O
of	O
the	O
aorta	O
,	O
the	O
proportion	O
of	O
macrophages	B-cell_type
with	O
SRA	B-protein
expression	O
was	O
highest	O
in	O
hypertensive	O
animals	O
on	O
a	O
high-cholesterol	O
diet	O
(	O
43.9+/-0.7	O
%	O
,	O
versus	O
12.0+/-2.0	O
%	O
for	O
normotensive	O
animals	O
on	O
a	O
high-cholesterol	O
diet	O
and	O
4.7+/-4.7	O
%	O
for	O
animals	O
on	O
standard	O
chow	O
;	O
P	O
<	O
0.001	O
)	O
.	O

CONCLUSIONS	O
:	O
Biomechanical	O
strain	O
induces	O
SRA	B-protein
expression	O
by	O
monocyte/macrophages	B-cell_type
,	O
suggesting	O
a	O
novel	O
mechanism	O
for	O
promotion	O
of	O
atherosclerosis	O
in	O
hypertensive	O
patients	O
.	O

-DOCSTART-	O

High	O
glucose	O
induces	O
MCP-1	O
expression	O
partly	O
via	O
tyrosine	O
kinase-AP-1	O
pathway	O
in	O
peritoneal	B-cell_type
mesothelial	I-cell_type
cells	I-cell_type
.	O

BACKGROUND	O
:	O
High	O
glucose	O
in	O
peritoneal	O
dialysis	O
solutions	O
has	O
been	O
implicated	O
in	O
the	O
pathogenesis	O
of	O
peritoneal	O
fibrosis	O
in	O
chronic	O
ambulatory	O
peritoneal	O
dialysis	O
(	O
CAPD	O
)	O
patients	O
.	O

However	O
,	O
the	O
mechanisms	O
are	O
not	O
very	O
clear	O
.	O

Peritoneal	B-cell_type
macrophages	I-cell_type
seem	O
to	O
participate	O
in	O
the	O
process	O
of	O
peritoneal	O
fibrosis	O
and	O
monocyte	B-protein
chemoattractant	I-protein
protein-1	I-protein
(	O
MCP-1	B-protein
)	O
plays	O
a	O
key	O
role	O
in	O
the	O
recruitment	O
of	O
monocytes	B-cell_type
toward	O
the	O
peritoneal	O
cavity	O
.	O

However	O
,	O
little	O
is	O
known	O
about	O
the	O
effect	O
of	O
high	O
glucose	O
on	O
MCP-1	B-protein
expression	O
and	O
its	O
signal	O
transduction	O
pathway	O
in	O
human	B-cell_type
peritoneal	I-cell_type
mesothelial	I-cell_type
cells	I-cell_type
.	O

METHODS	O
:	O
Mesothelial	B-cell_type
cells	I-cell_type
were	O
cultured	O
with	O
glucose	O
(	O
5	O
to	O
100	O
mmol/L	O
)	O
or	O
mannitol	O
chronically	O
for	O
up	O
to	O
seven	O
days	O
.	O

MCP-1	B-protein
expression	O
of	O
mRNA	O
and	O
protein	O
was	O
measured	O
by	O
Northern	O
blot	O
analysis	O
and	O
enzyme-linked	O
immunosorbent	O
assay	O
(	O
ELISA	O
)	O
.	O

Chemotactic	O
activity	O
of	O
high-glucose-conditioned	O
culture	O
supernatant	O
was	O
measured	O
by	O
chemotactic	O
assay	O
.	O

To	O
examine	O
the	O
roles	O
of	O
the	O
transcription	B-protein
factors	I-protein
activator	B-protein
protein-1	I-protein
(	O
AP-1	B-protein
)	O
and	O
nuclear	B-protein
factor-kappaB	I-protein
(	O
NF-kappaB	B-protein
)	O
,	O
electrophoretic	O
mobility	O
shift	O
assay	O
(	O
EMSA	O
)	O
was	O
performed	O
.	O

RESULTS	O
:	O
Glucose	O
induced	O
MCP-1	B-RNA
mRNA	I-RNA
expression	O
in	O
a	O
time-	O
and	O
dose-dependent	O
manner	O
.	O

MCP-1	B-protein
protein	I-protein
in	O
cell	O
culture	O
supernant	O
was	O
also	O
increased	O
.	O

Equivalent	O
concentrations	O
of	O
mannitol	O
had	O
no	O
significant	O
effect	O
.	O

High-glucose-conditioned	O
supernatant	O
possessed	O
an	O
increased	O
chemotactic	O
activity	O
for	O
monocytes	B-cell_type
,	O
which	O
was	O
neutralized	O
by	O
anti-MCP-1	B-protein
antibody	I-protein
.	O

EMSA	O
revealed	O
that	O
glucose	O
increased	O
the	O
AP-1	B-protein
binding	O
activity	O
in	O
a	O
time-	O
and	O
dose-dependent	O
manner	O
,	O
but	O
not	O
NF-kappaB	B-protein
.	O

Curcumin	O
,	O
an	O
inhibitor	O
of	O
AP-1	B-protein
,	O
dose-dependently	O
suppressed	O
the	O
induction	O
of	O
MCP-1	B-RNA
mRNA	I-RNA
by	O
high	O
glucose	O
.	O

Tyrosine	O
kinase	O
inhibitors	O
such	O
as	O
genistein	O
(	O
12.5	O
to	O
50	O
micromol/L	O
)	O
and	O
herbimycin	O
A	O
(	O
0.1	O
to	O
1	O
micromol/L	O
)	O
inhibited	O
the	O
high-glucose-induced	O
MCP-1	B-RNA
mRNA	I-RNA
expression	O
in	O
a	O
dose-dependent	O
manner	O
,	O
and	O
also	O
suppressed	O
the	O
high-glucose-induced	O
AP-1	B-protein
binding	O
activity	O
.	O

CONCLUSIONS	O
:	O
:	O
High	O
glucose	O
induced	O
mesothelial	O
MCP-1	B-protein
expression	O
partly	O
via	O
the	O
tyrosine	O
kinase-	O
AP-1	B-protein
pathway	O
.	O

-DOCSTART-	O

Signal	O
thresholds	O
and	O
modular	O
synergy	O
during	O
expression	O
of	O
costimulatory	O
molecules	O
in	O
B	B-cell_type
lymphocytes	I-cell_type
.	O

We	O
analyzed	O
intracellular	O
pathways	O
modulating	O
surface	O
densities	O
of	O
CD80	B-protein
and	O
CD86	B-protein
in	O
B	B-cell_type
cells	I-cell_type
activated	O
through	O
ligation	O
of	O
the	O
Ag	B-protein
receptor	I-protein
,	O
and	O
the	O
adhesion	B-protein
molecule	I-protein
CD54	B-protein
.	O

Whereas	O
B	B-protein
cell	I-protein
Ag	I-protein
receptor	I-protein
(	O
BCR	B-protein
)	O
cross-linking	O
alone	O
stimulated	O
increased	O
expression	O
of	O
CD86	B-protein
,	O
up-regulation	O
of	O
CD80	B-protein
required	O
dual	O
stimulation	O
with	O
anti-IgM	B-protein
and	O
anti-CD54	B-protein
.	O

The	O
principal	O
downstream	O
component	O
contributed	O
by	O
BCR	B-protein
signaling	O
,	O
toward	O
both	O
CD80	B-protein
and	O
CD86	B-protein
induction	O
,	O
was	O
the	O
elevated	O
concentration	O
of	O
free	O
cytoplasmic	O
Ca	O
(	O
2+	O
)	O
,	O
recruited	O
by	O
way	O
of	O
capacitative	O
influx	O
.	O

This	O
alone	O
was	O
sufficient	O
to	O
generate	O
an	O
increase	O
in	O
CD86	B-protein
levels	O
.	O

However	O
,	O
CD80	B-protein
enhancement	O
required	O
the	O
concerted	O
action	O
of	O
both	O
intracellular	O
Ca	O
(	O
2+	O
)	O
concentration	O
and	O
CD54-initiated	O
pathways	O
.	O

The	O
nexus	O
between	O
anti-IgM	O
and	O
anti-CD54	O
stimulation	O
,	O
in	O
the	O
context	O
of	O
CD80	B-protein
regulation	O
,	O
was	O
identified	O
to	O
involve	O
a	O
self-propagating	O
process	O
of	O
sequential	O
synergy	O
.	O

The	O
first	O
step	O
involved	O
amplified	O
accumulation	O
of	O
intracellular	O
cAMP	O
,	O
as	O
a	O
result	O
of	O
cross-talk	O
between	O
BCR	B-protein
-mobilized	O
Ca	O
(	O
2+	O
)	O
and	O
CD54-derived	O
signals	O
.	O

This	O
then	O
facilitated	O
a	O
second	O
synergistic	O
interaction	O
between	O
Ca	O
(	O
2+	O
)	O
and	O
cAMP	O
,	O
culminating	O
in	O
CD80	B-protein
expression	O
.	O

Our	O
findings	O
of	O
distinct	O
signal	O
transducer	O
requirements	O
,	O
with	O
the	O
added	O
consequences	O
of	O
cross-talk	O
,	O
offers	O
an	O
explanation	O
for	O
variable	O
modulation	O
of	O
costimulatory	O
molecule	O
expression	O
in	O
response	O
to	O
diverse	O
physiological	O
stimuli	O
.	O

Importantly	O
,	O
these	O
results	O
also	O
reveal	O
how	O
concentration	O
threshold	O
barriers	O
for	O
recruitment	O
of	O
individual	O
second	O
messengers	O
can	O
be	O
overcome	O
by	O
constructive	O
convergence	O
of	O
signaling	O
modules	O
.	O

-DOCSTART-	O

The	O
role	O
of	O
Epstein-Barr	O
virus	O
in	O
neoplastic	O
transformation	O
.	O

In	O
this	O
review	O
,	O
we	O
focus	O
on	O
new	O
data	O
from	O
basic	O
,	O
translational	O
and	O
clinical	O
research	O
relating	O
to	O
the	O
Epstein-Barr	O
virus	O
(	O
EBV	O
)	O
.	O

Beside	O
its	O
well-known	O
tropism	O
for	O
B	B-cell_type
lymphocytes	I-cell_type
and	O
epithelial	B-cell_type
cells	I-cell_type
,	O
EBV	O
also	O
infects	O
T	B-cell_type
lymphocytes	I-cell_type
,	O
monocytes	B-cell_type
and	O
granulocytes	B-cell_type
.	O

After	O
primary	O
infection	O
,	O
EBV	O
persists	O
throughout	O
the	O
life	O
span	O
in	O
resting	B-cell_type
memory	I-cell_type
B	I-cell_type
cells	I-cell_type
,	O
from	O
where	O
it	O
is	O
reactivated	O
upon	O
breakdown	O
of	O
cellular	O
immunity	O
.	O

In	O
the	O
process	O
of	O
neoplastic	O
transformation	O
,	O
the	O
EBV-encoded	B-DNA
latent	I-DNA
membrane	I-DNA
protein	I-DNA
1	I-DNA
(	I-DNA
LMP1	I-DNA
)	I-DNA
oncogene	I-DNA
represents	O
the	O
major	O
driving	O
force	O
.	O

LMP1	B-protein
acts	O
like	O
a	O
constitutively	B-protein
activated	I-protein
receptor	I-protein
of	O
the	O
tumor	B-protein
necrosis	I-protein
factor	I-protein
receptor	I-protein
family	I-protein
and	O
allows	O
the	O
amplification	O
or	O
bypassing	O
of	O
physiological	O
regulatory	O
signals	O
through	O
direct	O
and	O
indirect	O
interactions	O
with	O
proteins	O
of	O
the	O
tumor	B-protein
necrosis	I-protein
factor	I-protein
receptor-associated	I-protein
factor	I-protein
(	I-protein
TRAF	I-protein
)	I-protein
family	I-protein
.	O

TRAF2	B-protein
-mediated	O
NF-kappaB	B-protein
activation	O
,	O
AP-1	B-protein
induction	O
and	O
JAK3	B-protein
/	O
STAT	B-protein
activation	O
may	O
result	O
in	O
sustained	O
proliferation	O
leading	O
to	O
lymphoma	B-cell_line
.	O

The	O
ability	O
of	O
LMP1	B-protein
to	O
suppress	O
germinal	O
center	O
formation	O
and	O
its	O
capacity	O
to	O
mediate	O
its	O
own	O
transcriptional	O
activation	O
shed	O
new	O
light	O
on	O
the	O
pathogenesis	O
of	O
EBV-associated	O
latency	O
type	O
II	O
lymphoproliferations	O
like	O
Hodgkin	O
's	O
disease	O
and	O
angioimmunoblastic	O
lymphadenopathy	O
.	O

The	O
carboxy	B-protein
terminus	I-protein
of	O
LMP1	B-protein
is	O
also	O
a	O
reliable	O
marker	O
for	O
individual	O
EBV	O
strain	O
identification	O
and	O
thus	O
offers	O
new	O
possibilities	O
in	O
tracing	O
the	O
molecular	O
events	O
leading	O
to	O
posttransplant	O
lymphoproliferative	O
disorders	O
(	O
PTLDs	O
)	O
.	O

Cytotoxic	B-cell_type
T	I-cell_type
lymphocytes	I-cell_type
directed	O
against	O
well-characterized	O
epitopes	O
of	O
EBV	B-DNA
latency	I-DNA
genes	I-DNA
represent	O
an	O
already	O
successful	O
and	O
promising	O
therapeutic	O
approach	O
to	O
EBV-associated	B-cell_line
lymphomas	I-cell_line
,	O
in	O
particular	O
PTLDs	O

-DOCSTART-	O

Interferon-alpha	B-protein
drives	O
T	O
cell-mediated	O
immunopathology	O
in	O
the	O
intestine	O
.	O

The	O
ability	O
of	O
interferon	B-protein
(	I-protein
IFN	I-protein
)	I-protein
-alpha	I-protein
to	O
induce	O
autoimmunity	O
and	O
exacerbate	O
Th1	O
diseases	O
is	O
well	O
known	O
.	O

We	O
have	O
recently	O
described	O
enhanced	O
expression	O
of	O
IFN-alpha	B-protein
in	O
the	O
mucosa	O
of	O
patients	O
with	O
celiac	O
disease	O
(	O
CD	O
)	O
,	O
a	O
gluten-sensitive	O
Th1-mediated	O
enteropathy	O
,	O
characterized	O
by	O
villous	O
atrophy	O
and	O
crypt	O
cell	O
hyperplasia	O
.	O

Previous	O
studies	O
from	O
this	O
laboratory	O
have	O
shown	O
that	O
T	B-cell_type
cell	I-cell_type
activation	O
in	O
explant	O
cultures	O
of	O
human	O
fetal	O
gut	O
can	O
also	O
result	O
in	O
villous	O
atrophy	O
and	O
crypt	O
cell	O
hyperplasia	O
.	O

We	O
have	O
,	O
therefore	O
,	O
examined	O
changes	O
that	O
take	O
place	O
in	O
explant	O
cultures	O
of	O
human	O
fetal	O
gut	O
after	O
activation	O
of	O
T	B-cell_type
cells	I-cell_type
with	O
anti-CD3	B-protein
and/or	O
IFN-alpha	B-protein
.	O

We	O
show	O
that	O
activation	O
of	O
T	B-cell_type
cells	I-cell_type
with	O
anti-CD3	B-protein
alone	O
elicits	O
a	O
small	O
IFN-gamma	B-protein
and	O
TNF-alpha	B-protein
response	O
with	O
no	O
tissue	O
injury	O
.	O

Similarly	O
,	O
no	O
changes	O
are	O
seen	O
in	O
explants	O
cultured	O
with	O
IFN-alpha	B-protein
alone	O
.	O

However	O
,	O
addition	O
of	O
IFN-alpha	B-protein
with	O
anti-CD3	B-protein
results	O
in	O
enhanced	O
Th1	O
response	O
and	O
crypt	O
cell	O
hyperplasia	O
.	O

This	O
is	O
associated	O
with	O
enhanced	O
phosphorylation	O
of	O
STAT1	B-protein
,	O
STAT3	B-protein
,	O
and	O
Fyn	B-protein
,	I-protein
a	I-protein
Src	I-protein
homology	I-protein
tyrosine	I-protein
kinase	I-protein
,	O
which	O
interacts	O
with	O
both	O
TCR	O
and	O
IFN-alpha	O
signal	O
components	O
.	O

Together	O
these	O
data	O
indicate	O
that	O
IFN-alpha	B-protein
can	O
facilitate	O
activation	O
of	O
Th1-reactive	B-cell_type
cells	I-cell_type
in	O
the	O
gut	O
and	O
drive	O
immunopathology	O
.	O

-DOCSTART-	O

Suppression	O
of	O
tumor	B-protein
necrosis	I-protein
factor	I-protein
alpha	I-protein
production	O
by	O
cAMP	O
in	O
human	B-cell_type
monocytes	I-cell_type
:	O
dissociation	O
with	O
mRNA	O
level	O
and	O
independent	O
of	O
interleukin-10	B-protein
.	O

BACKGROUND	O
:	O
Elevation	O
of	O
cellular	O
cAMP	O
inhibits	O
lipopolysaccharide	O
(	O
LPS	O
)	O
-stimulated	O
tumor	B-protein
necrosis	I-protein
factor	I-protein
alpha	I-protein
(	O
TNF-alpha	B-protein
)	O
production	O
and	O
increases	O
the	O
expression	O
of	O
interleukin	B-protein
(	I-protein
IL	I-protein
)	I-protein
-10	I-protein
in	O
mononuclear	B-cell_type
cells	I-cell_type
.	O

TNF-alpha	B-DNA
gene	I-DNA
expression	O
obligates	O
activation	O
of	O
the	O
transcription	B-protein
factor	I-protein
nuclear	B-protein
factor	I-protein
kappaB	I-protein
(	O
NF-kappaB	B-protein
)	O
.	O

Exogenous	O
IL-10	B-protein
inhibits	O
NF-kappaB	B-protein
in	O
monocytes	B-cell_type
and	O
thus	O
attenuates	O
TNF-alpha	B-protein
production	O
.	O

We	O
examined	O
the	O
role	O
of	O
endogenous	O
IL-10	B-protein
in	O
the	O
regulation	O
of	O
NF-kappaB	B-protein
activation	O
and	O
TNF-alpha	B-protein
production	O
in	O
human	B-cell_type
monocytes	I-cell_type
by	O
cAMP	O
.	O

METHODS	O
:	O
Human	B-cell_type
monocytes	I-cell_type
were	O
stimulated	O
with	O
Escherichia	O
coli	O
LPS	O
(	O
100	O
ng/ml	O
)	O
with	O
and	O
without	O
forskolin	O
(	O
FSK	O
,	O
50	O
microM	O
)	O
or	O
dibutyryl	O
cyclic	O
AMP	O
(	O
dbcAMP	O
,	O
100	O
microM	O
)	O
.	O

Cytokine	B-protein
(	O
TNF-alpha	B-protein
and	O
IL-10	B-protein
)	O
release	O
was	O
measured	O
by	O
immunoassay	O
.	O

TNF-alpha	B-RNA
mRNA	I-RNA
was	O
measured	O
by	O
reverse	O
transcription	O
polymerase	O
chain	O
reaction	O
,	O
and	O
NF-kappaB	B-protein
DNA	O
binding	O
activity	O
was	O
assessed	O
by	O
gel	O
mobility	O
shift	O
assay	O
.	O

RESULTS	O
:	O
cAMP-elevating	O
agents	O
inhibited	O
LPS-stimulated	O
TNF-alpha	B-protein
release	O
(	O
0.77	O
+/-	O
0.13	O
ng/10	O
(	O
6	O
)	O
cells	O
in	O
LPS	O
+	O
dbcAMP	O
and	O
0.68	O
+/-	O
0.19	O
ng/10	O
(	O
6	O
)	O
cells	O
in	O
LPS	O
+	O
FSK	O
,	O
both	O
P	O
<	O
0.05	O
vs	O
1.61	O
+/-	O
0.34	O
ng/10	O
(	O
6	O
)	O
cells	O
in	O
LPS	O
alone	O
)	O
.	O

Conversely	O
,	O
cAMP	O
enhanced	O
LPS-stimulated	O
IL-10	B-protein
release	O
(	O
100	O
+/-	O
21.5	O
pg/10	O
(	O
6	O
)	O
cells	O
in	O
LPS	O
+	O
dbcAMP	O
and	O
110	O
+/-	O
25.2	O
pg/10	O
(	O
6	O
)	O
cells	O
in	O
LPS	O
+	O
FSK	O
,	O
both	O
P	O
<	O
0.05	O
vs	O
53.3	O
+/-	O
12.8	O
pg/10	O
(	O
6	O
)	O
cells	O
in	O
LPS	O
alone	O
)	O
.	O

Neither	O
TNF-alpha	B-RNA
mRNA	I-RNA
expression	O
nor	O
NF-kappaB	B-protein
activation	O
stimulated	O
by	O
LPS	O
was	O
inhibited	O
by	O
the	O
cAMP-elevating	O
agents	O
.	O

Neutralization	O
of	O
IL-10	B-protein
with	O
a	O
specific	O
antibody	B-protein
did	O
not	O
attenuate	O
the	O
effect	O
of	O
cAMP-elevating	O
agents	O
on	O
TNF-alpha	B-protein
production	O
.	O

CONCLUSION	O
:	O
The	O
results	O
indicate	O
that	O
cAMP	O
inhibits	O
LPS-stimulated	O
TNF-alpha	B-protein
production	O
through	O
a	O
posttranscriptional	O
mechanism	O
that	O
is	O
independent	O
of	O
endogenous	O
IL-10	B-protein
.	O

Copyright	O
2001	O
Academic	O
Press	O
.	O

-DOCSTART-	O

Activation	O
of	O
the	O
p21	B-DNA
(	I-DNA
CIP1/WAF1	I-DNA
)	I-DNA
promoter	I-DNA
by	O
bone	B-protein
morphogenetic	I-protein
protein-2	I-protein
in	O
mouse	O
B	B-cell_line
lineage	I-cell_line
cells	I-cell_line
.	O

BMPs	B-protein
exert	O
a	O
negative	O
growth	O
effect	O
on	O
various	O
types	O
of	O
cells	O
.	O

We	O
have	O
previously	O
reported	O
that	O
BMP-2	B-protein
inhibited	O
the	O
growth	O
of	O
HS-72	B-cell_line
mouse	I-cell_line
hybridoma	I-cell_line
cells	I-cell_line
by	O
inducing	O
p21	O
(	O
CIP1/WAF1	O
)	O
expression	O
.	O

In	O
the	O
present	O
study	O
,	O
we	O
demonstrated	O
that	O
BMP-2	B-protein
activated	O
the	O
mouse	O
p21	B-DNA
(	I-DNA
CIP1/WAF1	I-DNA
)	I-DNA
promoter	I-DNA
in	O
HS-72	B-cell_line
cells	I-cell_line
,	O
and	O
that	O
a	O
29-base	B-DNA
pair	I-DNA
(	I-DNA
b	I-DNA
)	I-DNA
region	I-DNA
of	O
the	O
promoter	O
(	O
-1928/-1900	O
relative	O
to	O
the	O
TATA	B-DNA
box	I-DNA
)	O
,	O
conserved	O
between	O
mice	O
and	O
humans	O
,	O
was	O
responsive	O
to	O
BMP-2	B-protein
as	O
well	O
as	O
expression	O
of	O
Smad1	B-protein
,	O
Smad4	B-protein
,	O
and	O
constitutively	O
active	O
mutants	O
of	O
BMP	B-protein
type	I-protein
I	I-protein
receptors	I-protein
.	O

Furthermore	O
,	O
an	O
oligonucleotide	O
containing	O
the	O
29-b	B-DNA
region	I-DNA
was	O
found	O
to	O
be	O
associated	O
with	O
Smad4	B-protein
and	O
phosphorylated	B-protein
Smad1	I-protein
in	O
the	O
nuclear	O
extract	O
of	O
BMP-2	B-protein
-stimulated	O
HS-72	B-cell_line
cells	I-cell_line
.	O

These	O
results	O
suggested	O
that	O
BMP-2	B-protein
might	O
activate	O
p21	O
(	O
CIP1/WAF1	O
)	O
transcription	O
by	O
inducing	O
a	O
binding	O
of	O
Smad4	B-protein
and	O
Smad1	B-protein
to	O
the	O
29-b	B-DNA
region	I-DNA
in	O
HS-72	B-cell_line
cells	I-cell_line
.	O

-DOCSTART-	O

Dendritic	B-cell_type
cell	I-cell_type
development	O
from	O
common	O
myeloid	B-cell_type
progenitors	I-cell_type
.	O

Dendritic	B-cell_type
cells	I-cell_type
(	O
DCs	B-cell_type
)	O
are	O
professional	O
antigen-presenting	B-cell_type
cells	I-cell_type
which	O
both	O
initiate	O
adaptive	O
immune	O
responses	O
and	O
control	O
tolerance	O
to	O
self-antigens	B-protein
.	O

It	O
has	O
been	O
suggested	O
that	O
these	O
different	O
effects	O
on	O
responder	O
cells	O
depend	O
on	O
subsets	O
of	O
DCs	B-cell_type
arising	O
from	O
either	O
myeloid	O
or	O
lymphoid	O
hematopoietic	O
origins	O
.	O

In	O
this	O
model	O
,	O
CD8	B-cell_type
alpha+	I-cell_type
Mac-1-	I-cell_type
DCs	I-cell_type
are	O
supposed	O
to	O
be	O
of	O
lymphoid	O
while	O
CD8	B-cell_type
alpha-	I-cell_type
Mac-1+	I-cell_type
DCs	I-cell_type
are	O
supposed	O
to	O
be	O
of	O
myeloid	O
origin	O
.	O

Here	O
we	O
summarize	O
our	O
findings	O
that	O
both	O
CD8	B-cell_type
alpha+	I-cell_type
and	I-cell_type
CD8	I-cell_type
alpha-	I-cell_type
DCs	I-cell_type
can	O
arise	O
from	O
clonogenic	B-cell_type
common	I-cell_type
myeloid	I-cell_type
progenitors	I-cell_type
(	O
CMPs	B-cell_type
)	O
in	O
both	O
thymus	O
and	O
spleen	O
.	O

Therefore	O
CD8	B-protein
alpha	I-protein
expression	O
DCs	B-cell_type
does	O
not	O
indicate	O
a	O
lymphoid	O
origin	O
and	O
differences	O
among	O
CD8	B-cell_type
alpha+	I-cell_type
and	I-cell_type
CD8	I-cell_type
alpha-	I-cell_type
DCs	I-cell_type
might	O
rather	O
reflect	O
maturation	O
status	O
than	O
ontogeny	O
.	O

On	O
the	O
basis	O
of	O
transplantation	O
studies	O
,	O
it	O
seems	O
likely	O
that	O
most	O
of	O
the	O
DCs	B-cell_type
in	O
secondary	O
lymphoid	O
organs	O
and	O
a	O
substantial	O
fraction	O
of	O
thymic	B-cell_type
DCs	I-cell_type
are	O
myeloid-derived	O
.	O

-DOCSTART-	O

Macrophage	O
stimulation	O
with	O
Murabutide	O
,	O
an	O
HIV-suppressive	O
muramyl	O
peptide	O
derivative	O
,	O
selectively	O
activates	O
extracellular	B-protein
signal-regulated	I-protein
kinases	I-protein
1	I-protein
and	I-protein
2	I-protein
,	O
C/EBPbeta	B-protein
and	O
STAT1	B-protein
:	O
role	O
of	O
CD14	B-protein
and	O
Toll-like	B-protein
receptors	I-protein
2	I-protein
and	I-protein
4	I-protein
.	O

The	O
smallest	O
unit	O
of	O
bacterial	O
peptidoglycans	O
known	O
to	O
be	O
endowed	O
with	O
biological	O
activities	O
is	O
muramyl	O
dipeptide	O
(	O
MDP	O
)	O
.	O

A	O
clinically	O
acceptable	O
synthetic	O
derivative	O
of	O
MDP	O
,	O
namely	O
murabutide	O
(	O
MB	O
)	O
,	O
has	O
been	O
found	O
to	O
present	O
interesting	O
pharmacological	O
properties	O
and	O
to	O
suppress	O
HIV-1	O
replication	O
in	O
monocyte-derived	B-cell_type
macrophages	I-cell_type
(	O
MDM	B-cell_type
)	O
.	O

We	O
have	O
addressed	O
the	O
signaling	O
events	O
activated	O
in	O
MDM	B-cell_type
following	O
stimulation	O
with	O
either	O
MB	O
or	O
the	O
potent	O
immunostimulant	O
LPS	O
.	O

We	O
also	O
examined	O
whether	O
signaling	O
by	O
muramyl	O
peptides	O
involves	O
the	O
use	O
of	O
cell	B-protein
surface	I-protein
receptors	I-protein
,	O
including	O
CD14	B-protein
and	O
Toll-like	B-protein
receptor	I-protein
2	I-protein
(	O
TLR2	B-protein
)	O
or	O
TLR4	B-protein
that	O
are	O
known	O
to	O
be	O
signal-transducing	B-protein
receptors	I-protein
for	O
other	O
bacterial	O
cell	O
wall	O
components	O
.	O

We	O
demonstrate	O
that	O
,	O
unlike	O
LPS	O
,	O
the	O
safe	O
immunomodulator	O
MB	O
selectively	O
activates	O
extracellular	B-protein
signal-regulated	I-protein
kinases	I-protein
(	I-protein
Erk	I-protein
)	I-protein
1/2	I-protein
,	O
in	O
the	O
absence	O
of	O
detectable	O
Jun	B-protein
N-terminal	I-protein
kinase	I-protein
(	O
JNK	B-protein
)	O
or	O
p38	B-protein
mitogen-activated	I-protein
kinase	I-protein
activation	O
.	O

Furthermore	O
,	O
STAT1	B-protein
activation	O
but	O
weak	O
or	O
no	O
activation	O
of	O
STAT3	B-protein
or	O
STAT5	B-protein
respectively	O
,	O
could	O
be	O
detected	O
in	O
MB-stimulated	B-cell_type
MDM	I-cell_type
.	O

Using	O
MonoMac6	B-cell_line
cells	I-cell_line
,	O
we	O
observed	O
high	O
C/EBPbeta	B-protein
and	O
AP-1	B-protein
but	O
weaker	O
and	O
transient	O
NF-kappaB	B-protein
activation	O
by	O
MB	O
.	O

Moreover	O
,	O
the	O
truncated	O
form	O
of	O
C/EBPbeta	B-protein
,	O
known	O
to	O
repress	O
HIV-1	O
transcription	O
,	O
was	O
detected	O
in	O
extracts	O
from	O
MB-treated	B-cell_line
THP-1	I-cell_line
cells	I-cell_line
.	O

Surprisingly	O
,	O
neither	O
MB	O
nor	O
MDP	O
were	O
able	O
to	O
transduce	O
signals	O
via	O
CD14	B-protein
and	O
TLR2	B-protein
or	I-protein
4	I-protein
.	O

These	O
findings	O
present	O
major	O
differences	O
in	O
the	O
early	O
cell	O
activation	O
process	O
between	O
LPS	O
and	O
muramyl	O
peptides	O
,	O
and	O
strongly	O
argue	O
for	O
the	O
implication	O
of	O
co-receptors	B-protein
other	O
than	O
TLR2	B-protein
and	O
TLR4	B-protein
in	O
mediating	O
the	O
signaling	O
events	O
induced	O
by	O
defined	O
subunits	O
of	O
bacterial	O
peptidoglycans	O
.	O

-DOCSTART-	O

Nuclear	B-protein
peroxisome	I-protein
proliferator-activated	I-protein
receptors	I-protein
alpha	I-protein
and	I-protein
gamma	I-protein
have	O
opposing	O
effects	O
on	O
monocyte	O
chemotaxis	O
in	O
endometriosis	O
.	O

The	O
peroxisome	B-protein
proliferator-activated	I-protein
receptors	I-protein
(	I-protein
PPARs	I-protein
)	I-protein
alpha	I-protein
and	I-protein
gamma	I-protein
are	O
nuclear	B-protein
receptors	I-protein
that	O
play	O
important	O
roles	O
in	O
inflammatory	O
diseases	O
like	O
ulcerative	O
colitis	O
and	O
arthritis	O
.	O

In	O
this	O
study	O
,	O
we	O
examined	O
the	O
possible	O
role	O
of	O
PPARs	B-protein
in	O
macrophage	O
attraction	O
into	O
the	O
peritoneal	O
cavity	O
of	O
patients	O
with	O
endometriosis	O
.	O

We	O
identified	O
PPAR-alpha	B-RNA
and	I-RNA
-gamma	I-RNA
messenger	O
RNA	O
by	O
RT-PCR	O
and	O
protein	O
by	O
immunoblotting	O
of	O
lysates	O
of	O
peritoneal	B-cell_type
macrophages	I-cell_type
and	O
monocytic	B-cell_line
U937	I-cell_line
cells	I-cell_line
.	O

Using	O
immunocytochemistry	O
,	O
we	O
localized	O
PPAR-alpha	B-protein
and	I-protein
-gamma	I-protein
within	O
the	O
nuclei	O
of	O
both	O
cell	O
types	O
.	O

Monocyte	O
chemotactic	O
activity	O
of	O
peritoneal	O
fluid	O
from	O
patients	O
with	O
endometriosis	O
was	O
quantified	O
in	O
Boyden	O
chambers	O
.	O

Migration	O
of	O
U937	B-cell_line
cells	I-cell_line
was	O
increased	O
by	O
WY	O
14643	O
and	O
reduced	O
by	O
rosiglitazone	O
.	O

Peritoneal	O
fluid	O
from	O
patients	O
with	O
endometriosis	O
activated	O
U937	B-cell_line
cells	I-cell_line
transiently	O
transfected	O
with	O
a	O
PPAR-alpha/GAL4	B-DNA
luciferase	I-DNA
reporter	I-DNA
.	O

By	O
contrast	O
,	O
peritoneal	O
fluid	O
did	O
not	O
cause	O
significant	O
activation	O
of	O
PPAR-gamma/GAL4	B-DNA
constructs	I-DNA
.	O

The	O
U937	B-cell_line
cells	I-cell_line
transiently	O
transfected	O
with	O
a	O
PPAR	B-DNA
response	I-DNA
element	I-DNA
luciferase	I-DNA
reporter	I-DNA
showed	O
disease	O
stage-dependent	O
up-regulation	O
when	O
treated	O
with	O
peritoneal	O
fluid	O
from	O
patients	O
with	O
endometriosis	O
.	O

Treatment	O
with	O
peritoneal	O
fluid	O
from	O
healthy	O
controls	O
down-regulated	O
PPAR	O
response	O
element	O
transactivation	O
.	O

We	O
conclude	O
that	O
peritoneal	O
fluid	O
of	O
endometriosis	O
patients	O
contains	O
activators	O
of	O
PPAR-alpha	B-protein
that	O
stimulate	O
macrophage	O
chemotaxis	O
.	O

Inhibitors	O
of	O
PPAR-alpha	B-protein
or	O
activators	O
of	O
PPAR-gamma	B-protein
could	O
be	O
developed	O
for	O
the	O
treatment	O
of	O
inflammation	O
associated	O
with	O
endometriosis	O
.	O

-DOCSTART-	O

Long-term-impaired	O
expression	O
of	O
nuclear	B-protein
factor-kappa	I-protein
B	I-protein
and	O
I	B-protein
kappa	I-protein
B	I-protein
alpha	I-protein
in	O
peripheral	B-cell_type
blood	I-cell_type
mononuclear	I-cell_type
cells	I-cell_type
of	O
trauma	O
patients	O
.	O

Nuclear	B-protein
factor	I-protein
(	I-protein
NF	I-protein
)	I-protein
-kappa	I-protein
B	I-protein
expression	O
and	O
dimer	O
characteristics	O
were	O
studied	O
in	O
peripheral	B-cell_type
blood	I-cell_type
mononuclear	I-cell_type
cells	I-cell_type
(	O
PBMCs	B-cell_type
)	O
of	O
major-trauma	O
patients	O
and	O
healthy	O
controls	O
.	O

Analysis	O
of	O
PBMCs	B-cell_type
on	O
days	O
1	O
,	O
3	O
,	O
5	O
,	O
and	O
10	O
after	O
trauma	O
revealed	O
that	O
expression	O
of	O
both	O
p65p50	B-protein
heterodimers	I-protein
and	O
p50p50	B-protein
homodimers	I-protein
was	O
significantly	O
reduced	O
compared	O
with	O
that	O
in	O
controls	O
.	O

In	O
vitro	O
lipopolysaccharide	O
(	O
LPS	O
)	O
stimulation	O
of	O
PBMCs	B-cell_type
induced	O
NF-kappa	O
B	O
translocation	O
.	O

However	O
,	O
throughout	O
the	O
survey	O
,	O
p65p50	O
activation	O
remained	O
significantly	O
lower	O
in	O
trauma	O
patients	O
than	O
in	O
controls	O
.	O

After	O
LPS	O
stimulation	O
in	O
vitro	O
,	O
the	O
p65p50/p50p50	O
ratio	O
was	O
significantly	O
lower	O
in	O
PBMCs	B-cell_type
from	O
trauma	O
patients	O
than	O
from	O
healthy	O
controls	O
.	O

The	O
ex	O
vivo	O
expression	O
of	O
I	B-protein
kappa	I-protein
B	I-protein
alpha	I-protein
was	O
higher	O
in	O
PBMCs	B-cell_type
of	O
controls	O
than	O
of	O
trauma	O
patients	O
.	O

LPS	O
did	O
not	O
induce	O
I	O
kappa	O
B	O
expression	O
in	O
PBMCs	B-cell_type
from	O
trauma	O
patients	O
,	O
but	O
strong	O
induction	O
was	O
obtained	O
with	O
staphylococci	O
,	O
suggesting	O
that	O
this	O
defect	O
is	O
not	O
universal	O
and	O
depends	O
on	O
the	O
nature	O
of	O
the	O
activating	O
signal	O
.	O

Although	O
no	O
direct	O
correlation	O
was	O
found	O
between	O
levels	O
of	O
interleukin-10	B-protein
or	O
transforming	B-protein
growth	I-protein
factor-beta	I-protein
and	O
NF-kappa	B-protein
B	I-protein
,	O
these	O
immunosuppressive	B-protein
cytokines	I-protein
were	O
significantly	O
elevated	O
in	O
trauma	O
patients	O
by	O
10	O
days	O
after	O
admission	O
.	O

The	O
long-term	O
low-basal	O
and	O
LPS-induced	O
nuclear	O
translocation	O
of	O
NF-kappa	B-protein
B	I-protein
recalled	O
long-term	O
immunoparalysis	O
observed	O
in	O
patients	O
with	O
severe	O
inflammatory	O
stress	O
such	O
as	O
trauma	O
.	O

-DOCSTART-	O

Hepatic	O
ischemia-reperfusion	O
injury	O
.	O

BACKGROUND	O
:	O
The	O
morbidity	O
associated	O
with	O
liver	O
transplantation	O
and	O
major	O
hepatic	O
resections	O
is	O
partly	O
a	O
result	O
of	O
ischemia-reperfusion	O
injury	O
.	O

DATA	O
SOURCES	O
:	O
The	O
entire	O
world	O
literature	O
on	O
the	O
subject	O
was	O
searched	O
via	O
Medline	O
.	O

Keywords	O
included	O
reperfusion	O
injury	O
,	O
transplantation	O
,	O
liver	O
resection	O
,	O
nitric	O
oxide	O
,	O
endothelin	O
,	O
cytokines	B-protein
,	O
Kupffer	B-cell_type
cells	I-cell_type
,	O
ischemic/ischaemic	O
preconditioning	O
,	O
and	O
nuclear	B-protein
factor-kappa	I-protein
B	I-protein
.	O

CONCLUSIONS	O
:	O
An	O
imbalance	O
between	O
endothelin	O
and	O
nitric	O
oxide	O
levels	O
results	O
in	O
failure	O
of	O
the	O
hepatic	O
microcirculation	O
at	O
the	O
onset	O
of	O
reperfusion	O
.	O

Activation	O
of	O
nuclear	B-protein
factor-kappa	I-protein
B	I-protein
in	O
the	O
liver	O
promotes	O
proinflammatory	B-protein
cytokine	I-protein
and	O
adhesion	B-protein
molecule	I-protein
synthesis	O
.	O

These	O
result	O
in	O
oxygen-derived	O
free	O
radical	O
production	O
and	O
neutrophil	O
recruitment	O
,	O
further	O
contributing	O
to	O
cellular	O
injury	O
.	O

Various	O
therapeutic	O
modalities	O
acting	O
on	O
the	O
above	O
mediators	O
have	O
been	O
successfully	O
used	O
to	O
attenuate	O
reperfusion	O
injury	O
in	O
animal	O
models	O
of	O
hepatic	O
transplantation	O
and	O
resection	O
.	O

Application	O
of	O
the	O
knowledge	O
gained	O
from	O
animal	O
models	O
of	O
hepatic	O
ischemia-reperfusion	O
to	O
the	O
clinical	O
setting	O
will	O
improve	O
the	O
outcome	O
of	O
hepatic	O
surgery	O
.	O

-DOCSTART-	O

Down-regulation	O
of	O
IL-12	B-DNA
p40	I-DNA
gene	I-DNA
in	O
Plasmodium	O
berghei-infected	O
mice	O
.	O

We	O
analyzed	O
the	O
mechanism	O
that	O
causes	O
suppression	O
of	O
IL-12	B-DNA
p40	I-DNA
gene	I-DNA
induction	O
during	O
Plasmodium	O
berghei	O
infection	O
.	O

Although	O
IL-12	B-protein
together	O
with	O
IFN-gamma	B-protein
plays	O
an	O
important	O
role	O
in	O
protection	O
against	O
pathogenic	O
infection	O
,	O
the	O
IL-12	B-protein
p70	I-protein
protein	I-protein
production	O
of	O
infected	B-cell_type
macrophages	I-cell_type
is	O
lower	O
than	O
that	O
by	O
the	O
uninfected	B-cell_type
macrophages	I-cell_type
.	O

We	O
showed	O
in	O
the	O
present	O
study	O
that	O
the	O
induction	O
of	O
IL-12	B-DNA
p40	I-DNA
gene	I-DNA
but	O
not	O
IL-12	B-DNA
p35	I-DNA
gene	I-DNA
in	O
macrophages	B-cell_type
of	O
P.	O
berghei-infected	O
mice	O
was	O
profoundly	O
inhibited	O
.	O

The	O
inhibition	O
was	O
induced	O
by	O
interaction	O
with	O
macrophages	B-cell_type
that	O
had	O
contacted	O
with	O
P.	B-cell_type
berghei-infected	I-cell_type
erythrocytes	I-cell_type
and	O
was	O
mediated	O
by	O
a	O
soluble	O
factor	O
,	O
IL-10	B-protein
.	O

There	O
was	O
comparable	O
activation	O
of	O
NF-kappaB	B-protein
in	O
uninfected	B-cell_type
and	I-cell_type
infected	I-cell_type
cells	I-cell_type
.	O

The	O
induction	O
of	O
IFN-regulatory	B-DNA
factor-1	I-DNA
gene	I-DNA
was	O
comparable	O
in	O
transcription	O
level	O
in	O
uninfected	B-cell_type
and	I-cell_type
infected	I-cell_type
cells	I-cell_type
,	O
while	O
the	O
unidentified	O
complex	O
formation	O
of	O
IFN-regulatory	B-protein
factor-1	I-protein
was	O
observed	O
in	O
infected	B-cell_type
cells	I-cell_type
.	O

Therefore	O
,	O
the	O
inhibition	O
of	O
the	O
IL-12	B-DNA
p40	I-DNA
gene	I-DNA
induction	O
appeared	O
to	O
be	O
regulated	O
at	O
transcriptional	O
regulation	O
level	O
of	O
the	O
gene	O
.	O

-DOCSTART-	O

The	O
Friend	B-protein
of	I-protein
GATA	I-protein
proteins	I-protein
U-shaped	O
,	O
FOG-1	B-protein
,	O
and	O
FOG-2	B-protein
function	O
as	O
negative	O
regulators	O
of	O
blood	O
,	O
heart	O
,	O
and	O
eye	O
development	O
in	O
Drosophila	O
.	O

Friend	B-protein
of	I-protein
GATA	I-protein
(	I-protein
FOG	I-protein
)	I-protein
proteins	I-protein
regulate	O
GATA	O
factor-activated	O
gene	O
transcription	O
.	O

During	O
vertebrate	O
hematopoiesis	O
,	O
FOG	B-protein
and	I-protein
GATA	I-protein
proteins	I-protein
cooperate	O
to	O
promote	O
erythrocyte	O
and	O
megakaryocyte	O
differentiation	O
.	O

The	O
Drosophila	B-protein
FOG	I-protein
homologue	I-protein
U-shaped	I-protein
(	O
Ush	B-protein
)	O
is	O
expressed	O
similarly	O
in	O
the	O
blood	O
cell	O
anlage	O
during	O
embryogenesis	O
.	O

During	O
hematopoiesis	O
,	O
the	O
acute	O
myeloid	O
leukemia	O
1	O
homologue	O
Lozenge	B-cell_type
and	I-cell_type
Glial	I-cell_type
cells	I-cell_type
missing	O
are	O
required	O
for	O
the	O
production	O
of	O
crystal	B-cell_type
cells	I-cell_type
and	O
plasmatocytes	B-cell_type
,	O
respectively	O
.	O

However	O
,	O
additional	O
factors	O
have	O
been	O
predicted	O
to	O
control	O
crystal	O
cell	O
proliferation	O
.	O

In	O
this	O
report	O
,	O
we	O
show	O
that	O
Ush	B-protein
is	O
expressed	O
in	O
hemocyte	B-cell_type
precursors	I-cell_type
and	O
plasmatocytes	B-cell_type
throughout	O
embryogenesis	O
and	O
larval	O
development	O
,	O
and	O
the	O
GATA	B-protein
factor	I-protein
Serpent	I-protein
is	O
essential	O
for	O
Ush	B-protein
embryonic	O
expression	O
.	O

Furthermore	O
,	O
loss	O
of	O
ush	O
function	O
results	O
in	O
an	O
overproduction	O
of	O
crystal	B-cell_type
cells	I-cell_type
,	O
whereas	O
forced	O
expression	O
of	O
Ush	B-protein
reduces	O
this	O
cell	O
population	O
.	O

Murine	B-protein
FOG-1	I-protein
and	I-protein
FOG-2	I-protein
also	O
can	O
repress	O
crystal	O
cell	O
production	O
,	O
but	O
a	O
mutant	O
version	O
of	O
FOG-2	B-protein
lacking	O
a	O
conserved	O
motif	O
that	O
binds	O
the	O
corepressor	B-protein
C-terminal	I-protein
binding	I-protein
protein	I-protein
fails	O
to	O
affect	O
the	O
cell	O
lineage	O
.	O

The	O
GATA	B-protein
factor	I-protein
Pannier	I-protein
(	O
Pnr	B-protein
)	O
is	O
required	O
for	O
eye	O
and	O
heart	O
development	O
in	O
Drosophila	O
.	O

When	O
Ush	B-protein
,	O
FOG-1	B-protein
,	O
FOG-2	B-protein
,	O
or	O
mutant	B-protein
FOG-2	I-protein
is	O
coexpressed	O
with	O
Pnr	B-protein
during	O
these	O
developmental	O
processes	O
,	O
severe	O
eye	O
and	O
heart	O
phenotypes	O
result	O
,	O
consistent	O
with	O
a	O
conserved	O
negative	O
regulation	O
of	O
Pnr	B-protein
function	O
.	O

These	O
results	O
indicate	O
that	O
the	O
fly	O
and	O
mouse	O
FOG	B-protein
proteins	I-protein
function	O
similarly	O
in	O
three	O
distinct	O
cellular	O
contexts	O
in	O
Drosophila	O
,	O
but	O
may	O
use	O
different	O
mechanisms	O
to	O
regulate	O
genetic	O
events	O
in	O
blood	O
vs	O
.	O
cardial	B-cell_line
or	I-cell_line
eye	I-cell_line
cell	I-cell_line
lineages	I-cell_line

-DOCSTART-	O

The	O
RING	B-protein
finger	I-protein
protein	I-protein
Siah-1	B-protein
regulates	O
the	O
level	O
of	O
the	O
transcriptional	B-protein
coactivator	I-protein
OBF-1	B-protein
.	O

The	O
transcriptional	B-protein
coactivator	I-protein
OBF-1	B-protein
,	O
which	O
interacts	O
with	O
Oct-1	B-protein
and	O
Oct-2	B-protein
and	O
the	O
octamer	B-DNA
site	I-DNA
DNA	I-DNA
,	O
has	O
been	O
shown	O
to	O
be	O
critical	O
for	O
development	O
of	O
a	O
normal	O
immune	O
response	O
and	O
the	O
formation	O
of	O
germinal	O
centers	O
in	O
secondary	O
lymphoid	O
organs	O
.	O

Here	O
we	O
have	O
identified	O
the	O
RING	B-protein
finger	I-protein
protein	I-protein
Siah-1	B-protein
as	O
a	O
protein	O
interacting	O
specifically	O
with	O
OBF-1	B-protein
.	O

This	O
interaction	O
is	O
mediated	O
by	O
the	O
C-terminal	B-protein
part	O
of	O
Siah-1	B-protein
and	O
by	O
residues	O
in	O
the	O
N-terminus	B-protein
of	O
OBF-1	B-protein
,	O
partly	O
distinct	O
from	O
the	O
residues	O
required	O
for	O
formation	O
of	O
a	O
complex	O
with	O
the	O
Oct	B-protein
POU	I-protein
domains	I-protein
and	O
the	O
DNA	O
.	O

Interaction	O
between	O
Siah-1	B-protein
and	O
OBF-1	B-protein
leads	O
to	O
downregulation	O
of	O
OBF-1	O
protein	O
level	O
but	O
not	O
mRNA	O
,	O
and	O
to	O
a	O
corresponding	O
reduction	O
in	O
octamer	O
site-dependent	O
transcription	O
activation	O
.	O

Inhibition	O
of	O
the	O
ubiquitin-proteasome	O
pathway	O
in	O
B	B-cell_type
cells	I-cell_type
leads	O
to	O
elevated	O
levels	O
of	O
OBF-1	B-protein
protein	I-protein
.	O

Furthermore	O
,	O
in	O
immunized	O
mice	O
,	O
OBF-1	B-protein
protein	I-protein
amounts	O
are	O
dramatically	O
increased	O
in	O
primary	B-cell_type
activated	I-cell_type
B	I-cell_type
cells	I-cell_type
,	O
without	O
concomitant	O
increase	O
in	O
OBF-1	B-RNA
mRNA	I-RNA
.	O

These	O
data	O
suggest	O
that	O
Siah-1	B-protein
is	O
part	O
of	O
a	O
novel	O
regulatory	O
loop	O
controlling	O
the	O
level	O
of	O
OBF-1	B-protein
protein	I-protein
in	O
B	B-cell_type
cells	I-cell_type
.	O

-DOCSTART-	O

The	O
beta-catenin	O
--	O
TCF-1	O
pathway	O
ensures	O
CD4	B-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
CD8	I-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
thymocyte	I-cell_type
survival	O
.	O

The	O
association	O
of	O
trans-acting	B-protein
T	I-protein
cell	I-protein
factors	I-protein
(	O
TCFs	B-protein
)	O
or	O
lymphoid	B-protein
enhancer	I-protein
factor	I-protein
1	I-protein
(	O
LEF-1	B-protein
)	O
with	O
their	O
coactivator	O
beta-catenin	O
mediates	O
transient	O
transcriptional	O
responses	O
to	O
extracellular	O
Wnt	O
signals	O
.	O

We	O
show	O
here	O
that	O
T	O
cell	O
maturation	O
depends	O
on	O
the	O
presence	O
of	O
the	O
beta-catenin	B-protein
--	I-protein
binding	I-protein
domain	I-protein
in	O
TCF-1	B-protein
.	O

This	O
domain	O
is	O
necessary	O
to	O
mediate	O
the	O
survival	O
of	O
immature	O
CD4	B-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
CD8	I-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
double-positive	I-cell_type
(	I-cell_type
DP	I-cell_type
)	I-cell_type
thymocytes	I-cell_type
.	O

Accelerated	O
spontaneous	O
thymocyte	O
death	O
in	O
the	O
absence	O
of	O
TCF-1	B-protein
correlates	O
with	O
aberrantly	O
low	O
expression	O
of	O
the	O
anti-apoptotic	B-protein
protein	I-protein
Bcl-x	B-protein
(	I-protein
L	I-protein
)	I-protein
.	O

Increasing	O
anti-apoptotic	O
effectors	O
in	O
thymocytes	B-cell_type
by	O
the	O
use	O
of	O
a	O
Bcl-2	B-DNA
transgene	I-DNA
rescued	O
TCF-1-deficient	B-cell_type
DP	I-cell_type
thymocytes	I-cell_type
from	O
apoptosis	O
.	O

Thus	O
,	O
TCF-1	B-protein
,	O
upon	O
association	O
with	O
beta-catenin	O
,	O
transiently	O
ensures	O
the	O
survival	O
of	O
immature	B-cell_type
T	I-cell_type
cells	I-cell_type
,	O
which	O
enables	O
them	O
to	O
generate	O
and	O
edit	O
T	B-protein
cell	I-protein
receptor	I-protein
(	I-protein
TCR	I-protein
)	I-protein
alpha	I-protein
chains	I-protein
and	O
attempt	O
TCR	B-protein
-mediated	O
positive	O
selection	O
.	O

-DOCSTART-	O

TRAIL/Apo2L	B-protein
ligand	I-protein
selectively	O
induces	O
apoptosis	O
and	O
overcomes	O
drug	O
resistance	O
in	O
multiple	O
myeloma	O
:	O
therapeutic	O
applications	O
.	O

Multiple	O
myeloma	O
(	O
MM	O
)	O
remains	O
incurable	O
and	O
novel	O
treatments	O
are	O
urgently	O
needed	O
.	O

Preclinical	O
in	O
vitro	O
and	O
in	O
vivo	O
evaluations	O
were	O
performed	O
to	O
assess	O
the	O
potential	O
therapeutic	O
applications	O
of	O
human	O
recombinant	O
tumor	B-protein
necrosis	I-protein
factor	I-protein
(	I-protein
TNF	I-protein
)	I-protein
-related	I-protein
apoptosis-inducing	I-protein
ligand/Apo2	I-protein
ligand	I-protein
(	O
TRAIL/Apo2L	B-protein
)	O
in	O
MM	O
.	O

TRAIL/Apo2L	B-protein
potently	O
induced	O
apoptosis	O
of	O
MM	B-cell_type
cells	I-cell_type
from	O
patients	O
and	O
the	O
majority	O
of	O
MM	B-cell_line
cell	I-cell_line
lines	I-cell_line
,	O
including	O
cells	O
sensitive	O
or	O
resistant	O
to	O
dexamethasone	O
(	O
Dex	O
)	O
,	O
doxorubicin	O
(	O
Dox	O
)	O
,	O
melphalan	O
,	O
and	O
mitoxantrone	O
.	O

TRAIL/Apo2L	B-protein
also	O
overcame	O
the	O
survival	O
effect	O
of	O
interleukin	B-protein
6	I-protein
on	O
MM	B-cell_type
cells	I-cell_type
and	O
did	O
not	O
affect	O
the	O
survival	O
of	O
peripheral	B-cell_type
blood	I-cell_type
and	I-cell_type
bone	I-cell_type
marrow	I-cell_type
mononuclear	I-cell_type
cells	I-cell_type
and	O
purified	B-cell_type
B	I-cell_type
cells	I-cell_type
from	O
healthy	O
donors	O
.	O

The	O
status	O
of	O
the	O
TRAIL	B-protein
receptors	I-protein
(	O
assessed	O
by	O
immunoblotting	O
and	O
flow	O
cytometry	O
)	O
could	O
not	O
predict	O
TRAIL	O
sensitivity	O
of	O
MM	B-cell_type
cells	I-cell_type
.	O

The	O
anti-MM	O
activity	O
of	O
TRAIL/Apo2L	B-protein
was	O
confirmed	O
in	O
nu/xid/bg	O
mice	O
xenografted	O
with	O
human	B-cell_type
MM	I-cell_type
cells	I-cell_type
;	O
TRAIL	B-protein
(	O
500	O
microg	O
intraperitoneally	O
daily	O
for	O
14	O
days	O
)	O
was	O
well	O
tolerated	O
and	O
significantly	O
suppressed	O
the	O
growth	O
of	O
plasmacytomas	B-cell_type
.	O

Dox	O
up-regulated	O
the	O
expression	O
of	O
the	O
TRAIL	B-protein
receptor	I-protein
death	B-protein
receptor	I-protein
5	I-protein
(	O
DR5	B-protein
)	O
and	O
synergistically	O
enhanced	O
the	O
effect	O
of	O
TRAIL	B-protein
not	O
only	O
against	O
MM	B-cell_type
cells	I-cell_type
sensitive	O
to	O
,	O
but	O
also	O
against	O
those	O
resistant	O
to	O
,	O
Dex-	O
or	O
Dox-induced	O
apoptosis	O
.	O

Nuclear	O
factor	O
(	O
NF	O
)	O
-kappaB	O
inhibitors	O
,	O
such	O
as	O
SN50	O
(	O
a	O
cell-permeable	O
inhibitor	O
of	O
the	O
nuclear	O
translocation	O
and	O
transcriptional	O
activity	O
of	O
NF-kappaB	B-protein
)	O
or	O
the	O
proteasome	O
inhibitor	O
PS-341	O
,	O
enhanced	O
the	O
proapoptotic	O
activity	O
of	O
TRAIL/Apo2L	B-protein
against	O
TRAIL-sensitive	B-cell_type
MM	I-cell_type
cells	I-cell_type
,	O
whereas	O
SN50	O
reversed	O
the	O
TRAIL	O
resistance	O
of	O
ARH-77	B-cell_type
and	I-cell_type
IM-9	I-cell_type
MM	I-cell_type
cells	I-cell_type
.	O

Importantly	O
,	O
normal	B-cell_type
B	I-cell_type
lymphocytes	I-cell_type
were	O
not	O
sensitized	O
to	O
TRAIL	B-protein
by	O
either	O
Dox	O
,	O
SN50	O
,	O
or	O
PS-341	O
.	O

These	O
preclinical	O
studies	O
suggest	O
that	O
TRAIL/Apo2L	B-protein
can	O
overcome	O
conventional	O
drug	O
resistance	O
and	O
provide	O
the	O
basis	O
for	O
clinical	O
trials	O
of	O
TRAIL	B-protein
-based	O
treatment	O
regimens	O
to	O
improve	O
outcome	O
in	O
patients	O
with	O
MM	O
.	O

(	O
Blood.	O
2001	O
;	O
98	O
:	O
795-804	O
)	O

-DOCSTART-	O

CD28	B-protein
costimulation	O
is	O
required	O
not	O
only	O
to	O
induce	O
IL-12	B-protein
receptor	I-protein
but	O
also	O
to	O
render	O
janus	B-protein
kinases/STAT4	I-protein
responsive	O
to	O
IL-12	B-protein
stimulation	O
in	O
TCR-triggered	B-cell_type
T	I-cell_type
cells	I-cell_type
.	O

The	O
activation	O
of	O
resting	B-cell_type
T	I-cell_type
cells	I-cell_type
for	O
the	O
acquisition	O
of	O
various	O
functions	O
depends	O
on	O
whether	O
CD28	O
costimulatory	O
signals	O
are	O
provided	O
upon	O
T	O
cell	O
receptor	O
stimulation	O
.	O

Here	O
,	O
we	O
investigated	O
how	O
CD28	O
costimulation	O
functions	O
to	O
allow	O
TCR	B-protein
-triggered	O
resting	B-cell_type
T	I-cell_type
cells	I-cell_type
to	O
acquire	O
IL-12	B-protein
responsiveness	O
.	O

When	O
T	B-cell_type
cells	I-cell_type
are	O
stimulated	O
with	O
low	O
doses	O
of	O
anti-CD3	B-protein
mAb	I-protein
,	O
CD28	O
costimulation	O
was	O
required	O
for	O
the	O
optimal	O
levels	O
of	O
IL-12	B-protein
receptor	I-protein
(	O
IL-12R	B-protein
)	O
expression	O
.	O

However	O
,	O
stimulation	O
of	O
T	B-cell_type
cells	I-cell_type
with	O
high	O
doses	O
of	O
anti-CD3	B-protein
alone	O
induced	O
comparable	O
levels	O
of	O
IL-12R	B-protein
expression	O
to	O
those	O
induced	O
upon	O
CD28	O
costimulation	O
.	O

Nevertheless	O
,	O
there	O
was	O
a	O
substantial	O
difference	O
in	O
IL-12	B-protein
responsiveness	O
between	O
these	O
two	O
groups	O
of	O
T	B-cell_type
cells	I-cell_type
:	O
compared	O
to	O
anti-CD28-costimulated	B-cell_type
T	I-cell_type
cells	I-cell_type
,	O
T	B-cell_type
cells	I-cell_type
that	O
were	O
not	O
costimulated	O
with	O
anti-CD28	B-protein
exhibited	O
decreased	O
levels	O
of	O
Janus	B-protein
kinases	I-protein
(	O
JAK	B-protein
)	O
JAK2/TYK2	B-protein
and	O
STAT4	B-protein
phosphorylation	O
and	O
IFN-y	O
production	O
following	O
IL-12	B-protein
stimulation	O
.	O

Importantly	O
,	O
STAT6	O
phosphorylation	O
following	O
IL-4	O
stimulation	O
was	O
not	O
decreased	O
in	O
anti-CD28-uncostimulated	B-cell_type
T	I-cell_type
cells	I-cell_type
.	O

These	O
resutls	O
indicate	O
that	O
CD28	O
costimulation	O
not	O
only	O
contributes	O
to	O
up-regulating	O
IL-12R	B-protein
expression	O
but	O
is	O
also	O
required	O
to	O
render	O
JAKs/STAT4	B-protein
responsive	O
to	O
IL-12	B-protein
stimulation	O
.	O

-DOCSTART-	O

Constitutively	O
activated	O
Akt-1	B-protein
is	O
vital	O
for	O
the	O
survival	O
of	O
human	B-cell_type
monocyte-differentiated	I-cell_type
macrophages	I-cell_type
.	O

Role	O
of	O
Mcl-1	B-protein
,	O
independent	O
of	O
nuclear	B-protein
factor	I-protein
(	I-protein
NF	I-protein
)	I-protein
-kappaB	I-protein
,	O
Bad	B-protein
,	O
or	O
caspase	O
activation	O
.	O

Recent	O
data	O
from	O
mice	O
deficient	O
for	O
phosphatase	B-protein
and	O
tensin	B-DNA
homologue	I-DNA
deleted	O
from	O
chromosome	B-DNA
10	I-DNA
or	O
src	B-protein
homology	I-protein
2	I-protein
domain-containing	I-protein
5	I-protein
'	I-protein
inositol	I-protein
phosphatase	I-protein
,	O
phosphatases	B-protein
that	O
negatively	O
regulate	O
the	O
phosphatidylinositol	B-protein
3-kinase	I-protein
(	O
PI3K	B-protein
)	O
pathway	O
,	O
revealed	O
an	O
increased	O
number	O
of	O
macrophages	B-cell_type
in	O
these	O
animals	O
,	O
suggesting	O
an	O
essential	O
role	O
for	O
the	O
PI3K	B-protein
pathway	O
for	O
macro-phage	B-cell_type
survival	O
.	O

Here	O
,	O
we	O
focused	O
on	O
the	O
role	O
of	O
the	O
PI3K-regulated	B-protein
serine/threonine	I-protein
kinase	I-protein
Akt-1	B-protein
in	O
modulating	O
macrophage	B-cell_type
survival	O
.	O

Akt-1	B-protein
was	O
constitutively	O
activated	O
in	O
human	B-cell_type
macrophages	I-cell_type
and	O
addition	O
of	O
the	O
PI3K	B-protein
inhibitor	O
,	O
LY294002	O
,	O
suppressed	O
the	O
activation	O
of	O
Akt-1	B-protein
and	O
induced	O
cell	O
death	O
.	O

Furthermore	O
,	O
suppression	O
of	O
Akt-1	B-protein
by	O
inhibition	O
of	O
PI3K	B-protein
or	O
a	O
dominant	B-protein
negative	I-protein
(	I-protein
DN	I-protein
)	I-protein
Akt-1	I-protein
resulted	O
in	O
loss	O
of	O
mitochondrial	O
transmembrane	O
potential	O
,	O
activation	O
of	O
caspases-9	B-protein
and	I-protein
-3	I-protein
,	O
and	O
DNA	O
fragmentation	O
.	O

The	O
effects	O
of	O
PI3K	B-protein
inhibition	O
were	O
reversed	O
by	O
the	O
ectopic	O
expression	O
of	O
constitutively	O
activated	O
Akt-1	B-protein
or	O
Bcl-x	B-protein
(	I-protein
L	I-protein
)	I-protein
.	O

Inhibition	O
of	O
PI3K	B-protein
/Akt-1	O
pathway	O
either	O
by	O
LY294002	O
or	O
DN	B-protein
Akt-1	I-protein
had	O
no	O
effect	O
on	O
the	O
constitutive	O
or	O
inducible	O
activation	O
of	O
nuclear	B-protein
factor	I-protein
(	I-protein
NF	I-protein
)	I-protein
-kappaB	I-protein
in	O
human	B-cell_type
macrophages	I-cell_type
.	O

However	O
,	O
after	O
inhibition	O
of	O
the	O
PI3K	B-protein
/	O
Akt-1	B-protein
pathway	O
,	O
a	O
marked	O
decrease	O
in	O
the	O
expression	O
of	O
the	O
antiapoptotic	B-protein
molecule	I-protein
Mcl-1	B-protein
,	O
but	O
not	O
other	O
Bcl-2	B-protein
family	I-protein
members	I-protein
was	O
observed	O
,	O
and	O
Mcl-1	B-protein
rescued	O
macrophages	B-cell_type
from	O
LY294002-induced	O
cell	O
death	O
.	O

Further	O
,	O
inhibition	O
of	O
Mcl-1	B-protein
by	O
antisense	O
oligonucleotides	O
,	O
also	O
resulted	O
in	O
macrophage	B-cell_type
apoptosis	O
.	O

Thus	O
,	O
our	O
findings	O
demonstrate	O
that	O
the	O
constitutive	O
activation	O
of	O
Akt-1	B-protein
regulates	O
macrophage	B-cell_type
survival	O
through	O
Mcl-1	B-protein
,	O
which	O
is	O
independent	O
of	O
caspases	B-protein
,	O
NF-kappaB	B-protein
,	O
or	O
Bad	B-protein
.	O

-DOCSTART-	O

GRbeta	O
expression	O
in	O
nasal	B-cell_type
polyp	I-cell_type
inflammatory	I-cell_type
cells	I-cell_type
and	O
its	O
relationship	O
to	O
the	O
anti-inflammatory	O
effects	O
of	O
intranasal	O
fluticasone	O
.	O

BACKGROUND	O
:	O
Nasal	O
polyposis	O
disease	O
is	O
an	O
inflammatory	O
disorder	O
with	O
intense	O
eosinophilic	O
infiltration	O
of	O
respiratory	O
mucosa	O
that	O
is	O
often	O
difficult	O
to	O
control	O
with	O
topical	O
steroids	O
.	O

Recent	O
evidence	O
suggests	O
that	O
overexpression	O
of	O
the	O
glucocorticoid	B-protein
receptor	I-protein
splice	I-protein
variant	I-protein
GRbeta	B-protein
in	O
inflammatory	B-cell_type
cells	I-cell_type
might	O
contribute	O
to	O
steroid	O
insensitivity	O
in	O
diseases	O
such	O
as	O
asthma	O
.	O

OBJECTIVE	O
:	O
The	O
purposes	O
of	O
this	O
investigation	O
were	O
to	O
determine	O
whether	O
nasal	B-cell_type
polyp	I-cell_type
(	I-cell_type
NP	I-cell_type
)	I-cell_type
inflammatory	I-cell_type
cells	I-cell_type
overexpress	O
GRbeta	B-protein
and	O
to	O
examine	O
whether	O
GRbeta	B-protein
overexpression	O
is	O
associated	O
with	O
insensitivity	O
to	O
the	O
potent	O
topical	O
steroid	O
fluticasone	O
propionate	O
(	O
FP	O
)	O
.	O

METHODS	O
:	O
Biopsies	O
were	O
obtained	O
from	O
10	O
subjects	O
with	O
NPs	O
before	O
and	O
4	O
weeks	O
after	O
treatment	O
with	O
intranasal	O
FP	O
.	O

Middle	O
turbinates	O
biopsies	O
from	O
6	O
healthy	O
,	O
nonallergic	O
subjects	O
served	O
as	O
normal	O
controls	O
.	O

Biopsies	O
were	O
immunostained	O
for	O
inflammatory	O
cell	O
markers	O
as	O
well	O
as	O
GRbeta	B-protein
and	O
probed	O
for	O
various	O
cytokine	B-RNA
mRNA	I-RNA
.	O

The	O
anti-inflammatory	O
response	O
to	O
FP	O
was	O
examined	O
in	O
relation	O
to	O
pretreatment	O
levels	O
of	O
GRbeta	O
expression	O
.	O

RESULTS	O
:	O
The	O
total	O
numbers	O
of	O
inflammatory	B-cell_type
cells	I-cell_type
were	O
increased	O
in	O
NPs	B-cell_type
.	O

The	O
percentage	O
of	O
inflammatory	B-cell_type
cells	I-cell_type
expressing	O
GRbeta	B-protein
was	O
also	O
increased	O
(	O
40.5	O
%	O
+/-	O
19.2	O
%	O
vs	O
16.1	O
%	O
+/-	O
4.0	O
%	O
,	O
P	O
=.009	O
)	O
.	O

GRbeta	B-protein
expression	O
in	O
NPs	B-cell_type
was	O
almost	O
exclusive	O
to	O
T	B-cell_type
lymphocytes	I-cell_type
,	O
eosinophils	B-cell_type
,	O
and	O
macrophages	B-cell_type
.	O

An	O
inverse	O
correlation	O
was	O
observed	O
between	O
the	O
baseline	O
inflammatory	O
cell	O
GRbeta	B-protein
expression	O
and	O
the	O
reduction	O
after	O
FP	O
treatment	O
in	O
EG2-positive	B-cell_type
eosinophils	I-cell_type
,	O
CD4-positive	B-cell_type
T	I-cell_type
lymphocytes	I-cell_type
,	O
endothelial	O
VCAM-1	B-protein
expression	O
,	O
and	O
IL-4	B-cell_type
mRNA-positive	I-cell_type
cells	I-cell_type
.	O

NPs	B-cell_type
that	O
were	O
``	O
FP-insensitive	O
''	O
in	O
terms	O
of	O
suppression	O
of	O
eosinophil	B-cell_type
numbers	O
(	O
major	B-protein
basic	I-protein
protein	I-protein
-positive	O
)	O
had	O
a	O
significantly	O
greater	O
percentage	O
of	O
GRbeta-positive	B-cell_type
inflammatory	I-cell_type
cells	I-cell_type
,	O
a	O
higher	O
ratio	O
of	O
GRbeta-positive/GRalpha-positive	B-cell_type
cells	I-cell_type
,	O
and	O
increased	O
numbers	O
of	O
GRbeta-positive	B-cell_type
eosinophils	I-cell_type
and	O
macrophages	B-cell_type
in	O
comparison	O
with	O
those	O
that	O
were	O
``	O
FP-sensitive.	O
''	O

``	B-cell_type
FP-insensitive	I-cell_type
''	I-cell_type
NPs	I-cell_type
also	O
demonstrated	O
a	O
higher	O
percentage	O
of	O
IL-5-positive	B-cell_type
inflammatory	I-cell_type
cells	I-cell_type
expressing	O
GRbeta	B-protein
before	O
and	O
after	O
FP	O
treatment	O
.	O

CONCLUSION	O
:	O
GRbeta	B-protein
expression	O
appears	O
to	O
be	O
a	O
marker	O
of	O
steroid	O
insensitivity	O
in	O
NPs	B-cell_type
.	O

Expression	O
of	O
GRbeta	B-protein
by	O
NP	B-cell_type
inflammatory	I-cell_type
cells	I-cell_type
,	O
particularly	O
T	B-cell_type
cells	I-cell_type
and	O
eosinophils	B-cell_type
,	O
might	O
render	O
them	O
resistant	O
to	O
suppression	O
by	O
topical	O
steroids	O
and	O
thereby	O
contribute	O
to	O
persistent	O
NP	B-cell_type
inflammation	O
.	O

-DOCSTART-	O

Transcriptional	O
regulation	O
of	O
galectin-10	B-protein
(	O
eosinophil	B-protein
Charcot-Leyden	I-protein
crystal	I-protein
protein	I-protein
)	O
:	O
a	O
GC	B-DNA
box	I-DNA
(	O
-44	O
to	O
-50	O
)	O
controls	O
butyric	O
acid	O
induction	O
of	O
gene	O
expression	O
.	O

Galectin-10	B-protein
(	O
gal-10	B-protein
,	O
also	O
known	O
as	O
Charcot-Leyden	B-protein
crystal	I-protein
protein	I-protein
)	O
is	O
a	O
member	O
of	O
the	O
galectin	B-protein
family	I-protein
of	O
beta-galactoside	B-protein
binding	I-protein
proteins	I-protein
that	O
is	O
expressed	O
uniquely	O
in	O
eosinophilic	B-cell_type
and	I-cell_type
basophilic	I-cell_type
leukocytes	I-cell_type
.	O

To	O
gain	O
a	O
better	O
understanding	O
of	O
galectin	O
gene	O
expression	O
,	O
we	O
present	O
an	O
analysis	O
of	O
the	O
transcriptional	O
regulation	O
of	O
the	O
gene	O
encoding	O
gal-10	B-protein
.	O

Analysis	O
of	O
the	O
minimal	B-DNA
promoter	I-DNA
revealed	O
nine	O
consensus-binding	B-DNA
sites	I-DNA
for	O
transcription	B-protein
factors	I-protein
,	O
including	O
several	O
that	O
are	O
also	O
found	O
in	O
the	O
minimal	B-DNA
promoters	I-DNA
of	O
galectins	B-DNA
-1	I-DNA
,	I-DNA
-2	I-DNA
,	I-DNA
and	I-DNA
-3	I-DNA
.	O

The	O
decrease	O
in	O
gal-10	O
promoter	O
activity	O
after	O
disruption	O
of	O
either	O
the	O
GC	B-DNA
box	I-DNA
(	O
-44	O
to	O
-50	O
)	O
or	O
the	O
Oct	B-DNA
site	I-DNA
(	O
-255	O
to	O
-261	O
)	O
suggests	O
that	O
these	O
sites	O
,	O
along	O
with	O
the	O
previously	O
characterized	O
GATA	B-DNA
and	I-DNA
EoTF	I-DNA
sites	I-DNA
,	O
are	O
necessary	O
for	O
full	O
promoter	O
activity	O
.	O

By	O
supershift	O
analysis	O
,	O
we	O
demonstrate	O
binding	O
of	O
the	O
transcription	B-protein
factors	I-protein
Sp1	B-protein
and	O
Oct1	B-protein
to	O
the	O
consensus	B-DNA
GC	I-DNA
box	I-DNA
and	O
the	O
Oct	B-DNA
site	I-DNA
,	O
respectively	O
.	O

Similar	O
to	O
gal-1	B-DNA
,	O
gal-10	O
expression	O
is	O
induced	O
by	O
butyric	O
acid	O
,	O
an	O
effect	O
that	O
is	O
lost	O
upon	O
ablation	O
of	O
the	O
GC	B-DNA
box	I-DNA
.	O

Additionally	O
,	O
we	O
demonstrate	O
AML3	O
binding	O
to	O
the	O
consensus	B-DNA
AML	I-DNA
site	I-DNA
and	O
YY1	B-protein
binding	I-protein
to	O
the	O
Inr	B-DNA
sequence	I-DNA
,	O
both	O
elements	O
functioning	O
as	O
silencers	O
in	O
the	O
gal-10	B-DNA
promoter	I-DNA
.	O

-DOCSTART-	O

Tumour-stromal	O
interactions	O
.	O

Role	O
of	O
the	O
stroma	O
in	O
mammary	O
development	O
.	O

Mammary	O
development	O
depends	O
on	O
branching	O
morphogenesis	O
,	O
namely	O
the	O
bifurcation	O
and	O
extension	O
of	O
ductal	O
growth	O
points	O
(	O
end	O
buds	O
)	O
and	O
secretory	O
lobules	O
into	O
a	O
more	O
or	O
less	O
fatty	O
stroma	O
.	O

Because	O
breast	O
carcinomas	O
are	O
overwhelmingly	O
ductal	O
in	O
origin	O
,	O
this	O
review	O
focuses	O
on	O
stromal	O
influences	O
guiding	O
postnatal	O
ductal	O
development	O
and	O
there	O
is	O
only	O
the	O
briefest	O
account	O
of	O
the	O
role	O
of	O
embryonic	O
stroma	O
(	O
mesenchyme	O
)	O
.	O

The	O
stroma	O
as	O
the	O
necessary	O
target	O
for	O
endocrine	O
mammogens	O
and	O
the	O
source	O
of	O
stimulatory	O
growth	O
factors	O
is	O
described	O
and	O
the	O
importance	O
of	O
mammary	O
epithelium-induced	O
modifications	O
of	O
the	O
periductal	O
stroma	O
is	O
emphasized	O
.	O

Evidence	O
is	O
presented	O
that	O
if	O
they	O
are	O
to	O
grow	O
,	O
end	O
buds	O
must	O
condition	O
proximal	O
fatty	O
stroma	O
by	O
recruiting	O
white	B-cell_type
blood	I-cell_type
cells	I-cell_type
as	O
well	O
as	O
inducing	O
stromal	O
cell	O
division	O
and	O
,	O
possibly	O
,	O
estrogen	B-protein
receptors	I-protein
.	O

The	O
induction	O
of	O
a	O
fibrous	O
stromal	O
tunic	O
around	O
the	O
end	O
bud	O
is	O
described	O
and	O
its	O
likely	O
role	O
as	O
a	O
complex	O
ductal	O
morphogen	O
is	O
discussed	O
;	O
a	O
possible	O
role	O
in	O
growth	O
inhibition	O
is	O
also	O
considered	O
.	O

Although	O
the	O
signals	O
governing	O
fibrotic	O
induction	O
,	O
ductal	O
morphogenesis	O
,	O
and	O
growth	O
inhibition	O
are	O
unknown	O
,	O
a	O
role	O
for	O
transforming	O
growth	B-protein
factor-beta	I-protein
is	O
highly	O
likely	O
and	O
is	O
discussed	O
.	O

Finally	O
,	O
a	O
need	O
for	O
new	O
conceptual	O
and	O
experimental	O
approaches	O
to	O
understanding	O
stromal-epithelial	O
signaling	O
is	O
discussed	O
.	O

-DOCSTART-	O

Adipophilin	B-protein
is	O
a	O
sensitive	O
marker	O
for	O
lipid	O
loading	O
in	O
human	B-cell_type
blood	I-cell_type
monocytes	I-cell_type
.	O

Adipophilin	B-protein
,	O
a	O
marker	O
of	O
lipid	O
accumulation	O
initially	O
described	O
in	O
adipocytes	B-cell_type
,	O
was	O
recently	O
shown	O
to	O
be	O
induced	O
in	O
macrophage	B-cell_type
foam	I-cell_type
cells	I-cell_type
.	O

We	O
found	O
that	O
even	O
freshly	B-cell_type
isolated	I-cell_type
blood	I-cell_type
monocytes	I-cell_type
express	O
adipophilin	B-protein
and	O
that	O
the	O
amount	O
of	O
adipophilin	B-protein
protein	I-protein
is	O
variable	O
in	O
monocytes	B-cell_type
from	O
different	O
healthy	O
individuals	O
.	O

However	O
,	O
the	O
physiological	O
expression	O
of	O
adipophilin	B-protein
does	O
not	O
correlate	O
with	O
the	O
levels	O
of	O
free	O
fatty	O
acids	O
,	O
cholesterylesters	O
or	O
free	O
cholesterol	O
.	O

Enzymatically	B-protein
modified	I-protein
low-density	I-protein
lipoprotein	I-protein
(	O
E-LDL	B-protein
)	O
induces	O
rapid	O
foam	O
cell	O
formation	O
in	O
monocytes	B-cell_type
and	O
upregulates	O
adipophilin	B-protein
mRNA	O
and	O
protein	O
within	O
2	O
h	O
of	O
incubation	O
.	O

This	O
rapid	O
induction	O
of	O
adipophilin	B-protein
is	O
accompanied	O
by	O
a	O
significant	O
increase	O
of	O
free	O
fatty	O
acids	O
in	O
monocytes	B-cell_type
incubated	O
with	O
E-LDL	B-protein
.	O

Adipophilin	B-protein
facilitates	O
the	O
uptake	O
of	O
free	O
fatty	O
acids	O
,	O
and	O
here	O
we	O
demonstrate	O
that	O
free	O
fatty	O
acids	O
increase	O
is	O
related	O
to	O
the	O
early	O
upregulation	O
of	O
adipophilin	O
expression	O
in	O
blood	B-cell_type
monocytes	I-cell_type
.	O

Fatty	O
acids	O
are	O
ligands	O
for	O
peroxisome	B-protein
proliferator-activated	I-protein
receptor-gamma	I-protein
(	O
PPARgamma	B-protein
)	O
,	O
and	O
the	O
upregulation	O
of	O
adipophilin	B-RNA
mRNA	I-RNA
by	O
PPARgamma	B-protein
agonists	O
like	O
15d-PGJ	O
(	O
2	O
)	O
and	O
ciglitazone	O
indicates	O
that	O
PPARgamma	B-protein
may	O
mediate	O
the	O
induction	O
of	O
adipophilin	O
expression	O
in	O
human	B-cell_type
blood	I-cell_type
monocytes	I-cell_type
.	O

-DOCSTART-	O

Constitutive	O
expression	O
of	O
MHC	B-DNA
class	I-DNA
II	I-DNA
genes	I-DNA
in	O
melanoma	B-cell_line
cell	I-cell_line
lines	I-cell_line
results	O
from	O
the	O
transcription	O
of	O
class	B-DNA
II	I-DNA
transactivator	I-DNA
abnormally	O
initiated	O
from	O
its	O
B	B-DNA
cell-specific	I-DNA
promoter	I-DNA
.	O

In	O
melanoma	B-cell_line
cell	I-cell_line
lines	I-cell_line
,	O
two	O
different	O
patterns	O
of	O
MHC	O
class	O
II	O
expression	O
have	O
been	O
described	O
,	O
either	O
an	O
IFN	O
gamma-inducible	O
expression	O
of	O
HLA-DR	B-DNA
and	O
HLA-DP	B-DNA
,	O
with	O
a	O
faint	O
or	O
null	O
expression	O
of	O
HLA-DQ	B-DNA
,	O
resembling	O
that	O
described	O
for	O
melanocytes	B-cell_type
,	O
or	O
a	O
constitutive	O
expression	O
,	O
i.e.	O
,	O
IFN-gamma	O
independent	O
,	O
of	O
all	O
three	O
HLA-D	B-DNA
isotypes	I-DNA
.	O

As	O
this	O
latter	O
phenotype	O
has	O
been	O
associated	O
with	O
a	O
more	O
rapid	O
progression	O
of	O
melanoma	O
tumors	O
,	O
we	O
have	O
analyzed	O
in	O
different	O
melanoma	B-cell_line
cell	I-cell_line
lines	I-cell_line
the	O
molecular	O
mechanisms	O
leading	O
to	O
this	O
abnormal	O
pattern	O
of	O
MHC	O
class	O
II	O
expression	O
.	O

In	O
agreement	O
with	O
the	O
evidence	O
of	O
a	O
coordinate	O
transcription	O
of	O
the	O
HLA-D	B-DNA
genes	I-DNA
in	O
these	O
cell	O
lines	O
,	O
we	O
have	O
shown	O
the	O
constitutive	O
expression	O
of	O
CIITA	B-RNA
(	I-RNA
class	I-RNA
II	I-RNA
transactivator	I-RNA
)	I-RNA
transcripts	I-RNA
,	O
CIITA	B-protein
being	O
known	O
as	O
the	O
master	O
switch	O
of	O
MHC	O
class	O
II	O
expression	O
.	O

Unexpectedly	O
,	O
these	O
transcripts	O
initiate	O
from	O
promoter	B-DNA
III	I-DNA
of	O
the	O
CIITA	B-DNA
gene	I-DNA
,	O
a	O
promoter	O
that	O
is	O
mainly	O
used	O
constitutively	O
in	O
B	B-cell_type
lymphocytes	I-cell_type
.	O

This	O
expression	O
was	O
further	O
shown	O
to	O
occur	O
through	O
factor	O
(	O
s	O
)	O
acting	O
on	O
the	O
enhancer	O
located	O
upstream	O
of	O
CIITA	B-DNA
promoter	I-DNA
III	I-DNA
,	O
which	O
was	O
previously	O
described	O
in	O
epithelioid	B-cell_type
cells	I-cell_type
as	O
an	O
IFN-gamma-response	O
sequence	O
.	O

The	O
hypothesis	O
of	O
a	O
general	O
abnormality	O
of	O
the	O
IFN-gamma	O
transduction	O
pathway	O
was	O
dismissed	O
.	O

Constitutive	O
transcription	O
of	O
CIITA	B-DNA
from	O
promoter	B-DNA
III	I-DNA
having	O
been	O
observed	O
in	O
unrelated	O
melanoma	B-cell_line
cell	I-cell_line
lines	I-cell_line
,	O
we	O
propose	O
the	O
hypothesis	O
that	O
this	O
phenomenon	O
might	O
not	O
be	O
a	O
random	O
event	O
,	O
but	O
could	O
be	O
linked	O
to	O
the	O
neoplasic	O
state	O
of	O
the	O
melanoma	B-cell_type
cells	I-cell_type

-DOCSTART-	O

Caspase	B-protein
-mediated	O
calcineurin	O
activation	O
contributes	O
to	O
IL-2	B-protein
release	O
during	O
T	O
cell	O
activation	O
.	O

Calcineurin	B-protein
,	O
a	O
Ca	B-protein
(	I-protein
2+	I-protein
)	I-protein
/calmodulin-dependent	I-protein
Ser/Thr	I-protein
phosphatase	I-protein
(	I-protein
protein	I-protein
phosphatase	I-protein
2B	I-protein
)	I-protein
,	O
plays	O
a	O
critical	O
role	O
in	O
IL-2	O
production	O
during	O
T	O
cell	O
activation	O
.	O

It	O
has	O
been	O
previously	O
reported	O
that	O
IL-2	B-protein
release	O
in	O
activated	O
Jurkat	B-cell_line
T	I-cell_line
requires	O
caspase-like	O
activity	O
(	O
Posmantur	O
et	O
al.	O
(	O
1998	O
)	O
Exp.	O
Cell.	O
Res.	O
244	O
,	O
302-309	O
)	O
.	O

We	O
report	O
here	O
that	O
the	O
60-kDa	B-protein
catalytic	I-protein
subunit	I-protein
of	O
calcineurin	B-protein
A	I-protein
(	O
Cn	B-protein
A	I-protein
)	O
was	O
partially	O
cleaved	O
to	O
a	O
45-kDa	B-protein
form	O
in	O
phytohemagglutinin	O
A	O
(	O
PHA	O
)	O
or	O
phorbol	O
ester	O
+	O
ionomycin	O
(	O
P	O
+	O
I	O
)	O
-activated	O
Jurkat	B-cell_line
cells	I-cell_line
.	O

In	O
parallel	O
,	O
proteolytic	O
activation	O
of	O
upstream	O
caspases	B-protein
(	O
caspase-8	O
and	O
-9	O
)	O
as	O
well	O
as	O
effector	B-protein
caspase-3	I-protein
was	O
also	O
observed	O
.	O

Cn	B-protein
A	I-protein
cleavage	O
was	O
caspase	O
mediated	O
,	O
since	O
it	O
was	O
inhibitable	O
by	O
pan-caspase	O
inhibitor	O
Cbz-Asp-CH	O
(	O
2	O
)	O
OC	O
(	O
O	O
)	O
-2	O
,	O
6-dichlorobenzene	O
(	O
Z-D-DCB	O
)	O
.	O

Cn	B-protein
A	I-protein
cleavage	O
was	O
also	O
observed	O
when	O
purified	O
calcineurin	B-protein
was	O
digested	O
in	O
vitro	O
with	O
caspase-3	B-protein
.	O

Truncated	B-protein
Cn	I-protein
A	I-protein
was	O
associated	O
with	O
enhanced	O
phosphatase	O
activity	O
and	O
reduced	O
calmodulin	O
sensitivity	O
.	O

Furthermore	O
,	O
in	O
PHA	O
or	O
P	O
+	O
I-activated	O
Jurkat	B-cell_line
cells	I-cell_line
,	O
dephosphorylation	O
of	O
calcineurin	B-protein
substrate	O
NFATc	B-protein
(	O
a	O
transcription	B-protein
factor	I-protein
known	O
to	O
be	O
involved	O
in	O
transactivation	O
of	O
the	O
IL-2	B-DNA
gene	I-DNA
)	O
,	O
was	O
also	O
suppressed	O
by	O
Z-D-DCB	O
.	O

Taken	O
together	O
,	O
our	O
results	O
suggest	O
that	O
caspase-mediated	O
cleavage	O
of	O
Cn	B-protein
A	I-protein
contributes	O
to	O
IL-2	O
production	O
during	O
T	O
cell	O
activation	O
.	O

Copyright	O
2001	O
Academic	O
Press	O
.	O

-DOCSTART-	O

Maturation	O
of	O
human	B-cell_type
dendritic	I-cell_type
cells	I-cell_type
as	O
sulfasalazine	O
target	O
.	O

AIM	O
:	O
Sulfasalazine	O
,	O
a	O
nonsteroidal	O
anti-inflammatory	O
drug	O
,	O
is	O
effective	O
in	O
treating	O
some	O
autoimmune	O
diseases	O
,	O
but	O
its	O
mechanism	O
of	O
action	O
is	O
unclear	O
.	O

To	O
determine	O
whether	O
dendritic	B-cell_type
cells	I-cell_type
could	O
be	O
a	O
possible	O
target	O
of	O
the	O
drug	O
,	O
we	O
studied	O
the	O
effects	O
of	O
sulfasalazine	O
and	O
its	O
metabolites	O
,	O
aminosalicylate	O
and	O
sulfapyridine	O
,	O
on	O
in	O
vitro	O
maturation	O
(	O
terminal	O
differentiation	O
)	O
of	O
human	B-cell_type
myeloid	I-cell_type
dendritic	I-cell_type
cells	I-cell_type
.	O

METHODS	O
:	O
We	O
prepared	O
immature	B-cell_type
dendritic	I-cell_type
cells	I-cell_type
by	O
incubating	O
CD14-positive	B-cell_type
cells	I-cell_type
in	O
the	O
presence	O
of	O
granulocyte-	B-protein
macrophage	I-protein
colony-stimulating	I-protein
factor	I-protein
and	O
interleukin	B-protein
(	I-protein
IL	I-protein
)	I-protein
-4	I-protein
.	O

The	O
cells	O
were	O
matured	O
by	O
addition	O
of	O
tumor	B-protein
necrosis	I-protein
factor	I-protein
(	I-protein
TNF	I-protein
)	I-protein
-a	I-protein
,	O
IL-1	B-protein
beta	I-protein
,	O
and	O
prostaglandin	B-protein
E2	I-protein
in	O
the	O
presence	O
of	O
sulfasalazine	O
or	O
its	O
metabolites	O
--	O
aminosalicylate	O
and	O
sulfapyridine	O
,	O
or	O
their	O
combinations	O
.	O

We	O
quantified	O
the	O
effect	O
of	O
drugs	O
on	O
the	O
dendritic	O
cell	O
characteristics	O
,	O
such	O
as	O
stimulation	O
of	O
autologous	O
and	O
allogeneic	O
pan-T	O
cell	O
proliferation	O
,	O
surface	O
marker	O
phenotype	O
,	O
IL-12	O
p40	O
subunit	O
secretion	O
,	O
and	O
activation	O
of	O
nuclear	B-protein
transcription	I-protein
factor	I-protein
(	I-protein
NF	I-protein
)	I-protein
-kappa	I-protein
B	I-protein
.	O

RESULTS	O
:	O
Dendritic	B-cell_type
cells	I-cell_type
treated	O
with	O
sulfasalazine	O
(	O
1.25	O
micromol/L	O
or	O
2.5	O
micromol/L	O
)	O
could	O
not	O
stimulate	O
T	B-cell_type
cells	I-cell_type
(	O
p	O
<	O
0.028	O
,	O
two-sided	O
paired	O
t-test	O
)	O
.	O

In	O
distinction	O
to	O
drug-free	O
maturing	O
dendritic	B-cell_type
cells	I-cell_type
,	O
2.5	O
micromol/L	O
sulfasalazine	O
upregulated	O
the	O
levels	O
of	O
CD14	B-protein
and	O
CD68	B-protein
and	O
downregulated	O
the	O
levels	O
of	O
CD40	B-protein
,	O
CD80	B-protein
,	O
and	O
CD83	B-protein
(	O
for	O
all	O
CD	O
markers	O
,	O
p	O
<	O
0.03	O
for	O
difference	O
between	O
measurements	O
in	O
the	O
absence	O
and	O
the	O
presence	O
of	O
sulfasalazine	O
)	O
.	O

From	O
concentration-dependent	O
changes	O
in	O
CD83	O
expression	O
,	O
we	O
found	O
an	O
apparent	O
ID50	O
>	O
>	O
1.5	O
micromol/L	O
sulfasalazine	O
.	O

The	O
apparent	O
ID50	O
value	O
for	O
aminosalicylate-inhibited	O
maturation	O
was	O
4	O
micromol/L	O
.	O

Sulfapyridine	O
had	O
no	O
effect	O
.	O

At	O
1.25	O
micromol/L	O
,	O
sulfasalazine	O
largely	O
inhibited	O
NF-kB	O
activation	O
in	O
dendritic	B-cell_type
cells	I-cell_type
.	O

CONCLUSION	O
:	O
Maturing	B-cell_type
human	I-cell_type
dendritic	I-cell_type
cells	I-cell_type
are	O
hundred-fold	O
more	O
sensitive	O
to	O
sulfasalazine	O
than	O
T	B-cell_type
cells	I-cell_type
and	O
NK	B-cell_type
cells	I-cell_type
and	O
the	O
most	O
sensitive	O
human	O
cells	O
described	O
so	O
far	O
.	O

Thus	O
,	O
dendritic	O
cell	O
maturation	O
is	O
an	O
important	O
target	O
of	O
sulfasalazine	O
.	O

Because	O
of	O
the	O
role	O
of	O
dendritic	B-cell_type
cells	I-cell_type
in	O
(	O
auto	O
)	O
immunity	O
,	O
inhibition	O
of	O
their	O
maturation	O
might	O
provide	O
a	O
target	O
for	O
further	O
optimization	O
of	O
sulfasalazine	O
therapy	O
.	O

-DOCSTART-	O

Defective	O
function	O
of	O
the	O
proteasome	B-protein
in	O
autoimmunity	O
:	O
involvement	O
of	O
impaired	O
NF-kappaB	O
activation	O
.	O

Type	O
1	O
diabetes	O
(	O
also	O
known	O
as	O
insulin-dependent	O
diabetes	O
mellitus	O
or	O
juvenile-onset	O
diabetes	O
)	O
is	O
usually	O
caused	O
by	O
T	O
cell-mediated	O
autoimmunity	O
,	O
with	O
a	O
prediabetic	O
state	O
characterized	O
by	O
the	O
production	O
of	O
autoantibodies	B-protein
specific	O
for	O
proteins	O
expressed	O
by	O
pancreatic	B-cell_type
beta	I-cell_type
cells	I-cell_type
.	O

The	O
nonobese	O
patient	O
with	O
diabetes	O
(	O
NOD	O
)	O
mouse	O
is	O
a	O
spontaneous	O
model	O
of	O
type	O
1	O
diabetes	O
with	O
a	O
strong	O
genetic	O
component	O
that	O
maps	O
to	O
the	O
major	B-DNA
histocompatibility	I-DNA
complex	I-DNA
(	I-DNA
MHC	I-DNA
)	I-DNA
region	I-DNA
of	O
the	O
genome	B-DNA
.	O

A	O
specific	O
proteasome	B-protein
defect	O
has	O
been	O
identified	O
in	O
NOD	O
mouse	O
in	O
select	O
lymphocytic	B-cell_line
and	I-cell_line
monocytic	I-cell_line
lineages	I-cell_line
that	O
results	O
from	O
down-regulation	O
of	O
expression	O
of	O
the	O
proteasome	B-protein
subunit	I-protein
LMP2	I-protein
,	O
which	O
is	O
encoded	O
by	O
a	O
gene	O
in	O
the	O
MHC	B-DNA
genomic	I-DNA
region	I-DNA
.	O

This	O
defect	O
prevents	O
the	O
proteolytic	O
processing	O
required	O
for	O
the	O
production	O
and	O
activation	O
of	O
the	O
transcription	B-protein
factor	I-protein
nuclear	I-protein
factor-kappaB	I-protein
(	O
NF-kappaB	B-protein
)	O
,	O
which	O
plays	O
important	O
roles	O
in	O
immune	O
and	O
inflammatory	O
responses	O
,	O
as	O
well	O
as	O
increases	O
the	O
susceptibility	O
of	O
the	O
affected	O
cells	O
to	O
apoptosis	O
induced	O
by	O
tumor	B-protein
necrosis	I-protein
factor-alpha	I-protein
(	O
TNF-alpha	B-protein
)	O
.	O

The	O
novel	O
role	O
of	O
the	O
proteasome	B-protein
in	O
dysfunction	O
in	O
autoimmunity	O
is	O
presented	O
and	O
documented	O
to	O
be	O
both	O
tissue	O
and	O
developmental	O
stage	O
specific	O
.	O

We	O
propose	O
a	O
role	O
of	O
the	O
proteasome	B-protein
as	O
a	O
step	O
in	O
disease	O
pathogenesis	O
and	O
tissue	O
targeting	O
.	O

-DOCSTART-	O

Down-regulation	O
of	O
TDT	O
transcription	O
in	O
CD4	B-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
CD8	I-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
thymocytes	I-cell_type
by	O
Ikaros	B-protein
proteins	I-protein
in	O
direct	O
competition	O
with	O
an	O
Ets	B-protein
activator	I-protein
.	O

Ikaros	B-protein
is	O
a	O
unique	O
regulator	O
of	O
lymphopoiesis	O
that	O
associates	O
with	O
pericentromeric	B-DNA
heterochromatin	I-DNA
and	O
has	O
been	O
implicated	O
in	O
heritable	O
gene	O
inactivation	O
.	O

Binding	O
and	O
competition	O
experiments	O
demonstrate	O
that	O
Ikaros	B-protein
dimers	I-protein
compete	O
with	O
an	O
Ets	B-protein
activator	I-protein
for	O
occupancy	O
of	O
the	O
lymphocyte-specific	B-DNA
TdT	I-DNA
promoter	I-DNA
.	O

Mutations	O
that	O
selectively	O
disrupt	O
Ikaros	O
binding	O
to	O
an	O
integrated	B-DNA
TdT	I-DNA
promoter	I-DNA
had	O
no	O
effect	O
on	O
promoter	O
function	O
in	O
a	O
CD4	B-cell_line
(	I-cell_line
+	I-cell_line
)	I-cell_line
CD8	I-cell_line
(	I-cell_line
+	I-cell_line
)	I-cell_line
thymocyte	I-cell_line
line	I-cell_line
.	O

However	O
,	O
these	O
mutations	O
abolished	O
down-regulation	O
on	O
differentiation	O
,	O
providing	O
evidence	O
that	O
Ikaros	B-protein
plays	O
a	O
direct	O
role	O
in	O
repression	O
.	O

Reduced	O
access	O
to	O
restriction	O
enzyme	O
cleavage	O
suggested	O
that	O
chromatin	O
alterations	O
accompany	O
down-regulation	O
.	O

The	O
Ikaros-dependent	O
down-regulation	O
event	O
and	O
the	O
observed	O
chromatin	O
alterations	O
appear	O
to	O
precede	O
pericentromeric	O
repositioning	O
.	O

Current	O
models	O
propose	O
that	O
the	O
functions	O
of	O
Ikaros	B-protein
should	O
be	O
disrupted	O
by	O
a	O
small	O
isoform	B-protein
that	O
retains	O
the	O
dimerization	B-protein
domain	I-protein
and	O
lacks	O
the	O
DNA-binding	B-protein
domain	I-protein
.	O

Surprisingly	O
,	O
in	O
the	O
CD4	B-cell_line
(	I-cell_line
+	I-cell_line
)	I-cell_line
CD8	I-cell_line
(	I-cell_line
+	I-cell_line
)	I-cell_line
thymocyte	I-cell_line
line	I-cell_line
,	O
overexpression	O
of	O
a	O
small	B-protein
Ikaros	I-protein
isoform	I-protein
had	O
no	O
effect	O
on	O
differentiation	O
or	O
on	O
the	O
pericentromeric	O
targeting	O
and	O
DNA-binding	O
properties	O
of	O
Ikaros	B-protein
.	O

Rather	O
,	O
the	O
small	O
isoform	B-protein
assembled	O
into	O
multimeric	B-protein
complexes	I-protein
with	O
DNA-bound	B-protein
Ikaros	I-protein
at	O
the	O
pericentromeric	O
foci	O
.	O

The	O
capacity	O
for	O
in	O
vivo	O
multimer	O
formation	O
suggests	O
that	O
interactions	O
between	O
Ikaros	B-protein
dimers	I-protein
bound	O
to	O
the	O
TdT	B-DNA
promoter	I-DNA
and	O
those	O
bound	O
to	O
pericentromeric	B-DNA
repeat	I-DNA
sequences	I-DNA
may	O
contribute	O
to	O
the	O
pericentromeric	O
repositioning	O
of	O
the	O
inactive	B-DNA
gene	I-DNA
.	O

-DOCSTART-	O

Type	B-protein
I	I-protein
interferons	I-protein
and	O
IL-12	B-protein
:	O
convergence	O
and	O
cross-regulation	O
among	O
mediators	O
of	O
cellular	O
immunity	O
.	O

Therapeutic	O
use	O
of	O
type	B-protein
I	I-protein
IFN	I-protein
(	O
IFN-alpha/beta	B-protein
)	O
has	O
become	O
common	O
.	O

Many	O
of	O
the	O
diverse	O
diseases	O
targeted	O
are	O
marked	O
by	O
pathogenetic	O
abnormalities	O
in	O
cell-mediated	O
immunity	O
(	O
CMI	O
)	O
,	O
these	O
cellular	O
immune	O
responses	O
either	O
causing	O
injury	O
to	O
the	O
host	O
,	O
lacking	O
sufficient	O
vigor	O
for	O
virus	O
or	O
tumor	O
clearance	O
,	O
or	O
both	O
.	O

In	O
general	O
,	O
therapeutic	O
efficacy	O
is	O
limited	O
.	O

It	O
is	O
thus	O
notable	O
that	O
the	O
pleiotropic	O
effects	O
of	O
type	B-protein
I	I-protein
IFN	I-protein
on	O
CMI	O
remain	O
poorly	O
understood	O
.	O

We	O
characterized	O
the	O
effects	O
of	O
type	B-protein
I	I-protein
IFN	I-protein
on	O
the	O
production	O
of	O
IL-12	B-protein
,	O
the	O
central	B-protein
immunoregulatory	I-protein
cytokine	I-protein
of	O
the	O
CD4	B-protein
(	I-protein
+	I-protein
)	I-protein
T	I-protein
cell	I-protein
arm	I-protein
of	O
CMI	O
.	O

We	O
show	O
that	O
type	B-protein
I	I-protein
IFN	I-protein
are	O
potent	O
inhibitors	O
of	O
IL-12	O
production	O
by	O
human	B-cell_type
monocytes/macrophages	I-cell_type
.	O

The	O
underlying	O
mechanism	O
involves	O
transcriptional	O
inhibition	O
of	O
the	O
IL-12p40	B-DNA
gene	I-DNA
,	O
marked	O
by	O
down-regulation	O
of	O
PU.1	O
binding	O
activity	O
at	O
the	O
upstream	B-DNA
Ets	I-DNA
site	I-DNA
of	O
the	O
IL-12p40	B-DNA
promoter	I-DNA
.	O

Type	B-protein
I	I-protein
IFN	I-protein
have	O
previously	O
been	O
shown	O
to	O
be	O
able	O
to	O
substitute	O
for	O
IL-12	B-protein
in	O
driving	O
IFN-gamma	O
production	O
from	O
T	B-cell_type
and	I-cell_type
NK	I-cell_type
cells	I-cell_type
.	O

The	O
ability	O
of	O
IFN-alpha/beta	B-protein
to	O
suppress	O
IL-12	O
production	O
while	O
up-regulating	O
IFN-gamma	O
production	O
suggests	O
a	O
possible	O
mechanistic	O
basis	O
for	O
the	O
difficulties	O
of	O
employing	O
these	O
cytokines	B-protein
in	O
diseases	O
involving	O
abnormalities	O
of	O
CMI	O
.	O

-DOCSTART-	O

Nuclear	B-protein
factor-kappaB	I-protein
suppressive	O
and	O
inhibitor-kappaB	B-protein
stimulatory	O
effects	O
of	O
troglitazone	O
in	O
obese	O
patients	O
with	O
type	O
2	O
diabetes	O
:	O
evidence	O
of	O
an	O
antiinflammatory	O
action	O
?	O

It	O
has	O
been	O
shown	O
recently	O
that	O
troglitazone	O
exerts	O
an	O
anti-inflammatory	O
effect	O
,	O
in	O
vitro	O
,	O
and	O
in	O
experimental	O
animals	O
.	O

To	O
test	O
these	O
properties	O
in	O
humans	O
,	O
we	O
investigated	O
the	O
effect	O
of	O
troglitazone	O
on	O
the	O
proinflammatory	B-protein
transcription	I-protein
factor	I-protein
nuclear	I-protein
factor-kappaB	I-protein
and	O
its	O
inhibitory	B-protein
protein	I-protein
IkappaB	I-protein
in	O
mononuclear	B-cell_type
cells	I-cell_type
(	O
MNC	B-cell_type
)	O
and	O
plasma	B-protein
soluble	I-protein
intracellular	I-protein
adhesion	I-protein
molecule-1	I-protein
,	O
monocyte	B-protein
chemoattractant	I-protein
protein-1	I-protein
,	O
plasminogen	B-protein
activator	I-protein
inhibitor-1	I-protein
,	O
and	O
C-reactive	B-protein
protein	I-protein
.	O

We	O
also	O
examined	O
the	O
effect	O
of	O
troglitazone	O
on	O
reactive	O
oxygen	O
species	O
generation	O
,	O
p47	O
(	O
phox	O
)	O
subunit	O
expression	O
,	O
9-hydroxyoctadecadienoic	O
acid	O
(	O
9-HODE	O
)	O
,	O
13-HODE	O
,	O
o-tyrosine	O
,	O
and	O
m-tyrosine	O
in	O
obese	O
patients	O
with	O
type	O
2	O
diabetes	O
.	O

Seven	O
obese	O
patients	O
with	O
type	O
2	O
diabetes	O
were	O
treated	O
with	O
troglitazone	O
(	O
400	O
mg/day	O
)	O
for	O
4	O
weeks	O
.	O

Blood	O
samples	O
were	O
obtained	O
at	O
weekly	O
intervals	O
.	O

Nuclear	B-protein
factor-kappaB	I-protein
binding	O
activity	O
in	O
MNC	B-cell_type
nuclear	O
extracts	O
was	O
significantly	O
inhibited	O
after	O
troglitazone	O
treatment	O
at	O
week	O
1	O
and	O
continued	O
to	O
be	O
inhibited	O
up	O
to	O
week	O
4	O
.	O

On	O
the	O
other	O
hand	O
,	O
IkappaB	O
protein	O
levels	O
increased	O
significantly	O
after	O
troglitazone	O
treatment	O
at	O
week	O
1	O
,	O
and	O
this	O
increase	O
persisted	O
throughout	O
the	O
study	O
.	O

Plasma	B-protein
monocyte	I-protein
chemoattractant	I-protein
protein-1	I-protein
and	O
soluble	B-protein
intracellular	I-protein
adhesion	I-protein
molecule-1	I-protein
concentrations	O
did	O
not	O
decrease	O
significantly	O
after	O
troglitazone	O
treatment	O
,	O
although	O
there	O
was	O
a	O
trend	O
toward	O
inhibition	O
.	O

Reactive	O
oxygen	O
species	O
generation	O
by	O
polymorphonuclear	B-cell_type
cells	I-cell_type
and	O
MNC	B-cell_type
,	O
p47	B-protein
(	I-protein
phox	I-protein
)	I-protein
subunit	I-protein
protein	I-protein
quantities	O
,	O
plasminogen	B-protein
activator	I-protein
inhibitor-1	I-protein
,	O
and	O
C-reactive	B-protein
protein	I-protein
levels	O
decreased	O
significantly	O
after	O
troglitazone	O
intake	O
.	O

13-HODE/linoleic	O
acid	O
and	O
9-HODE/linoleic	O
acid	O
ratios	O
also	O
decreased	O
after	O
troglitazone	O
intake	O
.	O

However	O
,	O
o-tyrosine/phenylalanine	O
and	O
m-tyrosine/phenylalanine	O
ratios	O
did	O
not	O
change	O
significantly	O
.	O

These	O
data	O
show	O
that	O
troglitazone	O
has	O
profound	O
antiinflammatory	O
effects	O
in	O
addition	O
to	O
antioxidant	O
effects	O
in	O
obese	O
type	O
2	O
diabetics	O
;	O
these	O
effects	O
may	O
be	O
relevant	O
to	O
the	O
recently	O
described	O
beneficial	O
antiatherosclerotic	O
effects	O
of	O
troglitazone	O
at	O
the	O
vascular	O
level	O
.	O

-DOCSTART-	O

OX40	O
stimulation	O
by	O
gp34/OX40	B-protein
ligand	I-protein
enhances	O
productive	O
human	O
immunodeficiency	O
virus	O
type	O
1	O
infection	O
.	O

OX40	B-protein
is	O
a	O
member	O
of	O
the	O
tumor	B-protein
necrosis	I-protein
factor	I-protein
(	I-protein
TNF	I-protein
)	I-protein
receptor	I-protein
superfamily	I-protein
and	O
known	O
to	O
be	O
an	O
important	O
costimulatory	B-protein
molecule	I-protein
expressed	O
on	O
activated	B-cell_type
T	I-cell_type
cells	I-cell_type
.	O

To	O
investigate	O
the	O
role	O
of	O
costimulation	O
of	O
OX40	B-protein
in	O
human	O
immunodeficiency	O
virus	O
type	O
1	O
(	O
HIV-1	O
)	O
infection	O
by	O
its	O
natural	O
ligand	O
,	O
gp34	B-protein
,	O
the	O
OX40-transfected	B-cell_line
ACH-2	I-cell_line
cell	I-cell_line
line	I-cell_line
,	I-cell_line
ACH-2/OX40	I-cell_line
,	O
chronically	O
infected	O
with	O
HIV-1	O
,	O
was	O
cocultured	O
with	O
paraformaldehyde	O
(	O
PFA	O
)	O
-fixed	O
gp34-transfected	B-cell_line
mouse	I-cell_line
cell	I-cell_line
line	I-cell_line
,	I-cell_line
SV-T2/gp34	I-cell_line
.	O

The	O
results	O
showed	O
that	O
HIV-1	O
production	O
was	O
strongly	O
induced	O
.	O

This	O
was	O
followed	O
by	O
apparent	O
apoptosis	O
,	O
and	O
both	O
processes	O
were	O
specifically	O
inhibited	O
by	O
the	O
gp34	B-protein
-specific	O
neutralizing	B-protein
monoclonal	I-protein
antibody	I-protein
5A8	I-protein
.	O

Endogenous	O
TNF	B-protein
alpha	I-protein
(	O
TNF-alpha	B-protein
)	O
and	O
TNF-beta	B-protein
production	O
were	O
not	O
involved	O
in	O
the	O
enhanced	O
HIV-1	O
production	O
.	O

Furthermore	O
,	O
enhanced	O
HIV-1	O
transcription	O
in	O
gp34	B-protein
-stimulated	O
ACH-2/OX40	B-cell_line
cells	I-cell_line
was	O
dependent	O
on	O
the	O
kappa	B-DNA
B	I-DNA
site	I-DNA
of	O
the	O
HIV-1	B-DNA
long	I-DNA
terminal	I-DNA
repeat	I-DNA
,	O
and	O
the	O
OX40-gp34	O
interaction	O
activated	O
NF-kappa	B-protein
B	I-protein
consisting	O
of	O
p50	B-protein
and	I-protein
p65	I-protein
subunits	I-protein
.	O

When	O
primary	O
activated	O
CD4	B-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
T	I-cell_type
cells	I-cell_type
acutely	O
infected	O
with	O
HIV-1	O
(	O
NL4-3	O
)	O
(	O
CXCR4-using	B-cell_line
T-cell-line-tropic	I-cell_line
)	O
were	O
cocultured	O
with	O
PFA-fixed	O
gp34	B-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
human	I-cell_type
T-cell	I-cell_type
leukemia	O
virus	O
type	O
1-bearing	O
MT-2	B-cell_line
cells	I-cell_line
or	O
SV-T2/gp34	B-cell_line
cells	I-cell_line
,	O
HIV-1	O
production	O
was	O
also	O
markedly	O
enhanced	O
.	O

The	O
enhancement	O
was	O
again	O
significantly	O
inhibited	O
by	O
5A8	B-protein
.	O

The	O
present	O
study	O
first	O
shows	O
that	O
OX40-gp34	O
interaction	O
stimulates	O
HIV-1	O
expression	O
and	O
suggests	O
that	O
OX40	O
triggering	O
by	O
gp34	B-protein
may	O
play	O
an	O
important	O
role	O
in	O
enhancing	O
HIV-1	O
production	O
in	O
both	O
acutely	O
and	O
latently	O
infected	O
CD4	B-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
T	I-cell_type
cells	I-cell_type
in	O
vivo	O
.	O

-DOCSTART-	O

Troglitazone	O
,	O
a	O
PPARgamma	O
ligand	O
,	O
inhibits	O
osteopontin	O
gene	O
expression	O
in	O
human	B-cell_type
monocytes/macrophage	I-cell_type
THP-1	I-cell_type
cells	I-cell_type
.	O

Peroxizome	B-protein
proliferator-activated	I-protein
receptor-gamma	I-protein
(	O
PPARgamma	B-protein
)	O
is	O
a	O
member	O
of	O
the	O
nuclear	B-protein
receptor	I-protein
family	I-protein
of	O
transcription	B-protein
factors	I-protein
that	O
regulate	O
adipocyte	O
differentiation	O
.	O

Recent	O
studies	O
indicate	O
that	O
liganded	B-protein
PPARgamma	I-protein
not	O
only	O
promotes	O
differentiation	O
but	O
also	O
inhibits	O
the	O
activation	O
of	O
macrophages	B-cell_type
.	O

Osteopontin	B-protein
,	O
a	O
component	O
of	O
extracellular	O
matrix	O
,	O
is	O
synthesized	O
by	O
macrophages	B-cell_type
in	O
atherosclerotic	O
plaques	O
.	O

In	O
this	O
study	O
,	O
we	O
examined	O
whether	O
PPARgamma	B-protein
ligand	O
regulates	O
osteopontin	O
gene	O
expression	O
in	O
THP-1	B-cell_line
cells	I-cell_line
,	O
a	O
cell	O
line	O
derived	O
from	O
human	B-cell_type
monocytic	I-cell_type
leukemia	I-cell_type
cells	I-cell_type
which	O
can	O
differentiate	O
to	O
macrophage	B-cell_type
upon	O
stimulation	O
with	O
phorbol	O
ester	O
PMA	O
.	O

Northern	O
blot	O
analysis	O
showed	O
that	O
osteopontin	O
expression	O
is	O
markedly	O
induced	O
in	O
response	O
to	O
PMA	O
.	O

Troglitazone	O
,	O
a	O
PPARgamma	B-protein
ligand	O
,	O
dramatically	O
attenuated	O
the	O
PMA-induced	O
osteopontin	O
expression	O
.	O

Transient	O
transfection	O
assays	O
of	O
the	O
human	B-DNA
osteopontin	I-DNA
promoter/luciferase	I-DNA
construct	I-DNA
which	O
contains	O
a	O
5'-flanking	B-DNA
region	I-DNA
between	O
-1500	O
and	O
+87	O
relative	O
to	O
the	O
transcription	B-DNA
start	I-DNA
site	I-DNA
demonstrate	O
that	O
either	O
treatment	O
with	O
troglitazone	O
or	O
cotransfection	O
of	O
PPARgamma	B-DNA
expression	I-DNA
vector	I-DNA
inhibits	O
osteopontin	O
promoter	O
activity	O
.	O

These	O
data	O
indicate	O
that	O
troglitazone	O
reduces	O
osteopontin	O
gene	O
expression	O
at	O
transcriptional	O
level	O
through	O
PPARgamma	B-protein
activation	O
,	O
and	O
suggest	O
the	O
role	O
of	O
troglitazone	O
in	O
inhibiting	O
the	O
ability	O
of	O
macrophages	B-cell_type
to	O
produce	O
extracellular	O
matrix	O
,	O
which	O
is	O
particularly	O
relevant	O
to	O
atherosclerotic	O
plaque	O
formation	O
.	O

-DOCSTART-	O

Transcription	B-protein
factor	I-protein
NF-kappa	I-protein
B	I-protein
regulates	O
Ig	B-protein
lambda	I-protein
light	O
chain	O
gene	O
rearrangement	O
.	O

The	O
tissue-	O
and	O
stage-specific	O
assembly	O
of	O
Ig	B-DNA
and	I-DNA
TCR	I-DNA
genes	I-DNA
is	O
mediated	O
by	O
a	O
common	O
V	B-protein
(	I-protein
D	I-protein
)	I-protein
J	I-protein
recombinase	I-protein
complex	I-protein
in	O
precursor	B-cell_type
lymphocytes	I-cell_type
.	O

Directed	O
alterations	O
in	O
the	O
accessibility	O
of	O
V	B-DNA
,	I-DNA
D	I-DNA
,	I-DNA
and	I-DNA
J	I-DNA
gene	I-DNA
segments	I-DNA
target	O
the	O
recombinase	B-protein
to	O
specific	O
Ag	B-DNA
receptor	I-DNA
loci	I-DNA
.	O

Accessibility	O
within	O
a	O
given	O
locus	B-DNA
is	O
regulated	O
by	O
the	O
functional	O
interaction	O
of	O
transcription	B-protein
factors	I-protein
with	O
cognate	B-DNA
enhancer	I-DNA
elements	I-DNA
and	O
correlates	O
with	O
the	O
transcriptional	O
activity	O
of	O
unrearranged	O
gene	B-DNA
segments	I-DNA
.	O

As	O
demonstrated	O
in	O
our	O
prior	O
studies	O
,	O
rearrangement	O
of	O
the	O
Igkappa	B-DNA
locus	I-DNA
is	O
regulated	O
by	O
the	O
inducible	B-protein
transcription	I-protein
factor	I-protein
NF-kappaB	B-protein
.	O

In	O
contrast	O
to	O
the	O
Igkappa	B-DNA
locus	I-DNA
,	O
known	O
transcriptional	B-DNA
control	I-DNA
elements	I-DNA
in	O
the	O
Iglambda	B-DNA
locus	I-DNA
lack	O
functional	O
NF-kappaB	B-DNA
binding	I-DNA
sites	I-DNA
.	O

Consistent	O
with	O
this	O
observation	O
,	O
the	O
expression	O
of	O
assembled	O
Iglambda	B-DNA
genes	I-DNA
in	O
mature	B-cell_type
B	I-cell_type
cells	I-cell_type
has	O
been	O
shown	O
to	O
be	O
NF-kappaB	B-protein
independent	O
.	O

Nonetheless	O
,	O
we	O
now	O
show	O
that	O
specific	O
repression	O
of	O
NF-kappaB	B-protein
inhibits	O
germline	O
transcription	O
and	O
recombination	O
of	O
Iglambda	B-DNA
gene	I-DNA
segments	I-DNA
in	O
precursor	B-cell_type
B	I-cell_type
cells	I-cell_type
.	O

Molecular	O
analyses	O
indicate	O
that	O
the	O
block	O
in	O
NF-kappaB	B-protein
impairs	O
Iglambda	O
rearrangement	O
at	O
the	O
level	O
of	O
recombinase	B-protein
accessibility	O
.	O

In	O
contrast	O
,	O
the	O
activities	O
of	O
known	O
Iglambda	B-DNA
promoter	I-DNA
and	O
enhancer	B-DNA
elements	I-DNA
are	O
unaffected	O
in	O
the	O
same	O
cellular	O
background	O
.	O

These	O
findings	O
expand	O
the	O
range	O
of	O
NF-kappaB	B-protein
action	O
in	O
precursor	B-cell_type
B	I-cell_type
cells	I-cell_type
beyond	O
Igkappa	B-protein
to	O
include	O
the	O
control	O
of	O
recombinational	O
accessibility	O
at	O
both	O
L	B-DNA
chain	I-DNA
loci	I-DNA
.	O

Moreover	O
,	O
our	O
results	O
strongly	O
suggest	O
the	O
existence	O
of	O
a	O
novel	O
Iglambda	B-DNA
regulatory	I-DNA
element	I-DNA
that	O
is	O
either	O
directly	O
or	O
indirectly	O
activated	O
by	O
NF-kappaB	B-protein
during	O
the	O
early	O
stages	O
of	O
B	O
cell	O
development	O
.	O

-DOCSTART-	O

Transcription	B-protein
factor	I-protein
STAT5A	B-protein
is	O
a	O
substrate	O
of	O
Bruton	B-protein
's	I-protein
tyrosine	I-protein
kinase	I-protein
in	O
B	B-cell_type
cells	I-cell_type
.	O

STAT5A	B-protein
is	O
a	O
molecular	O
regulator	O
of	O
proliferation	O
,	O
differentiation	O
,	O
and	O
apoptosis	O
in	O
lymphohematopoietic	B-cell_type
cells	I-cell_type
.	O

Here	O
we	O
show	O
that	O
STAT5A	B-protein
can	O
serve	O
as	O
a	O
functional	O
substrate	O
of	O
Bruton	B-protein
's	I-protein
tyrosine	I-protein
kinase	I-protein
(	O
BTK	B-protein
)	O
.	O

Purified	B-protein
recombinant	I-protein
BTK	I-protein
was	O
capable	O
of	O
directly	O
binding	O
purified	B-protein
recombinant	I-protein
STAT5A	I-protein
with	O
high	O
affinity	O
(	O
K	O
(	O
d	O
)	O
=	O
44	O
nm	O
)	O
,	O
as	O
determined	O
by	O
surface	O
plasmon	O
resonance	O
using	O
a	O
BIAcore	O
biosensor	O
system	O
.	O

BTK	B-protein
was	O
also	O
capable	O
of	O
tyrosine-phosphorylating	O
ectopically	O
expressed	O
recombinant	B-protein
STAT5A	I-protein
on	O
Tyr	O
(	O
694	O
)	O
both	O
in	O
vitro	O
and	O
in	O
vivo	O
in	O
a	O
Janus	B-protein
kinase	I-protein
3	I-protein
-independent	O
fashion	O
.	O

BTK	B-protein
phosphorylated	O
the	O
Y665F	B-protein
,	I-protein
Y668F	I-protein
,	I-protein
and	I-protein
Y682F	I-protein
,	I-protein
Y683F	I-protein
mutants	I-protein
but	O
not	O
the	O
Y694F	B-protein
mutant	I-protein
of	O
STAT5A	B-protein
.	O

STAT5A	O
mutations	O
in	O
the	O
Src	B-protein
homology	I-protein
2	I-protein
(	I-protein
SH2	I-protein
)	I-protein
and	I-protein
SH3	I-protein
domains	I-protein
did	O
not	O
alter	O
the	O
BTK	B-protein
-mediated	O
tyrosine	O
phosphorylation	O
.	O

Recombinant	B-protein
BTK	I-protein
proteins	I-protein
with	O
mutant	O
pleckstrin	O
homology	O
,	O
SH2	B-protein
,	I-protein
or	I-protein
SH3	I-protein
domains	I-protein
were	O
capable	O
of	O
phosphorylating	O
STAT5A	B-protein
,	O
whereas	O
recombinant	B-protein
BTK	I-protein
proteins	I-protein
with	O
SH1/kinase	O
domain	O
mutations	O
were	O
not	O
.	O

In	O
pull-down	O
experiments	O
,	O
only	O
full-length	B-protein
BTK	I-protein
and	O
its	O
SH1/kinase	B-protein
domain	I-protein
(	O
but	O
not	O
the	O
pleckstrin	O
homology	O
,	O
SH2	B-protein
,	I-protein
or	I-protein
SH3	I-protein
domains	I-protein
)	O
were	O
capable	O
of	O
binding	O
STAT5A	B-protein
.	O

Ectopically	O
expressed	O
BTK	B-protein
kinase	I-protein
domain	I-protein
was	O
capable	O
of	O
tyrosine-phosphorylating	O
STAT5A	B-protein
both	O
in	O
vitro	O
and	O
in	O
vivo	O
.	O

BTK	B-protein
-mediated	O
tyrosine	O
phosphorylation	O
of	O
ectopically	O
expressed	O
wild	O
type	O
(	O
but	O
not	O
Tyr	B-protein
(	I-protein
694	I-protein
)	I-protein
mutant	I-protein
)	O
STAT5A	B-protein
enhanced	O
its	O
DNA	O
binding	O
activity	O
.	O

In	O
BTK	B-protein
-competent	O
chicken	O
B	B-cell_type
cells	I-cell_type
,	O
anti-IgM-stimulated	O
tyrosine	O
phosphorylation	O
of	O
STAT5	B-protein
protein	I-protein
was	O
prevented	O
by	O
pretreatment	O
with	O
the	O
BTK	B-protein
inhibitor	O
LFM-A13	O
but	O
not	O
by	O
pretreatment	O
with	O
the	O
JAK3	O
inhibitor	O
HI-P131	O
.	O

B	O
cell	O
antigen	O
receptor	O
ligation	O
resulted	O
in	O
enhanced	O
tyrosine	O
phosphorylation	O
of	O
STAT5	B-protein
in	O
BTK	B-protein
-deficient	O
chicken	O
B	B-cell_type
cells	I-cell_type
reconstituted	O
with	O
wild	B-protein
type	I-protein
human	I-protein
BTK	I-protein
but	O
not	O
in	O
BTK	B-protein
-deficient	O
chicken	O
B	B-cell_type
cells	I-cell_type
reconstituted	O
with	O
kinase-inactive	B-protein
mutant	I-protein
BTK	I-protein
.	O

Similarly	O
,	O
anti-IgM	O
stimulation	O
resulted	O
in	O
enhanced	O
tyrosine	O
phosphorylation	O
of	O
STAT5A	B-protein
in	O
BTK	B-protein
-competent	O
B	B-cell_type
cells	I-cell_type
from	O
wild	O
type	O
mice	O
but	O
not	O
in	O
BTK	B-protein
-deficient	O
B	B-cell_type
cells	I-cell_type
from	O
XID	O
mice	O
.	O

In	O
contrast	O
to	O
B	B-cell_type
cells	I-cell_type
from	O
XID	O
mice	O
,	O
B	B-cell_type
cells	I-cell_type
from	O
JAK3	O
knockout	O
mice	O
showed	O
a	O
normal	O
STAT5A	O
phosphorylation	O
response	O
to	O
anti-IgM	O
stimulation	O
.	O

These	O
findings	O
provide	O
unprecedented	O
experimental	O
evidence	O
that	O
BTK	B-protein
plays	O
a	O
nonredundant	O
and	O
pivotal	O
role	O
in	O
B	O
cell	O
antigen	O
receptor-mediated	O
STAT5A	O
activation	O
in	O
B	B-cell_type
cells	I-cell_type

-DOCSTART-	O

Role	O
of	O
T-bet	B-protein
in	O
commitment	O
of	O
TH1	B-cell_type
cells	I-cell_type
before	O
IL-12	B-protein
-dependent	O
selection	O
.	O

How	O
cytokines	B-protein
control	O
differentiation	O
of	O
helper	B-cell_type
T	I-cell_type
(	I-cell_type
TH	I-cell_type
)	I-cell_type
cells	I-cell_type
is	O
controversial	O
.	O

We	O
show	O
that	O
T-bet	B-protein
,	O
without	O
apparent	O
assistance	O
from	O
interleukin	B-protein
12	I-protein
(	I-protein
IL-12	I-protein
)	I-protein
/	O
STAT4	B-protein
,	O
specifies	O
TH1	B-cell_type
effector	O
fate	O
by	O
targeting	O
chromatin	O
remodeling	O
to	O
individual	O
interferon-gamma	B-DNA
(	I-DNA
IFN-gamma	I-DNA
)	I-DNA
alleles	I-DNA
and	O
by	O
inducing	O
IL-12	B-DNA
receptor	I-DNA
beta2	I-DNA
expression	O
.	O

Subsequently	O
,	O
it	O
appears	O
that	O
IL-12	B-protein
/	O
STAT4	B-protein
serves	O
two	O
essential	O
functions	O
in	O
the	O
development	O
of	O
TH1	B-cell_type
cells	I-cell_type
:	O
as	O
growth	B-protein
signal	I-protein
,	O
inducing	O
survival	O
and	O
cell	O
division	O
;	O
and	O
as	O
trans-activator	B-protein
,	O
prolonging	O
IFN-gamma	B-protein
synthesis	O
through	O
a	O
genetic	O
interaction	O
with	O
the	O
coactivator	B-protein
,	O
CREB-binding	B-protein
protein	I-protein
.	O

These	O
results	O
suggest	O
that	O
a	O
cytokine	B-protein
does	O
not	O
simply	O
induce	O
TH	O
fate	O
choice	O
but	O
instead	O
may	O
act	O
as	O
an	O
essential	O
secondary	O
stimulus	O
that	O
mediates	O
selective	O
survival	O
of	O
a	O
lineage	O
.	O

-DOCSTART-	O

Stat6	B-protein
is	O
necessary	O
and	O
sufficient	O
for	O
IL-4	O
's	O
role	O
in	O
Th2	O
differentiation	O
and	O
cell	O
expansion	O
.	O

IL-4	B-protein
plays	O
a	O
critical	O
role	O
in	O
the	O
differentiation	O
of	O
T	O
CR-stimulated	B-cell_type
naive	I-cell_type
CD4	I-cell_type
T	I-cell_type
cells	I-cell_type
to	O
the	O
Th2	O
phenotype	O
.	O

In	O
response	O
to	O
IL-4	B-protein
,	O
the	O
IL-4R	B-protein
activates	O
a	O
set	O
of	O
phosphotyrosine	B-protein
binding	I-protein
domain-containing	I-protein
proteins	I-protein
,	O
including	O
insulin	B-protein
receptor	I-protein
substrate	I-protein
1/2	I-protein
,	O
Shc	B-protein
,	O
and	O
IL-4R	B-protein
interacting	I-protein
protein	I-protein
,	O
as	O
well	O
as	O
Stat6	B-protein
.	O

Stat6	B-protein
has	O
been	O
shown	O
to	O
be	O
required	O
for	O
Th2	O
differentiation	O
.	O

To	O
determine	O
the	O
roles	O
of	O
the	O
phosphotyrosine	B-protein
binding	I-protein
adaptors	I-protein
in	O
Th2	O
differentiation	O
,	O
we	O
prepared	O
a	O
retrovirus	O
containing	O
a	O
mutant	O
of	O
the	O
human	B-protein
(	I-protein
h	I-protein
)	I-protein
IL-4R	I-protein
alpha-chain	I-protein
,	O
Y497F	B-protein
,	O
which	O
is	O
unable	O
to	O
recruit	O
these	O
adaptors	O
.	O

The	O
mutant	B-protein
hIL-4Ralpha	I-protein
,	O
as	O
well	O
as	O
the	O
wild-type	B-protein
(	I-protein
WT	I-protein
)	I-protein
hIL-4Ralpha	I-protein
,	O
was	O
introduced	O
into	O
naive	B-cell_type
CD4	I-cell_type
T	I-cell_type
cells	I-cell_type
.	O

Upon	O
hIL-4	O
stimulation	O
,	O
Y497F	B-protein
worked	O
as	O
well	O
as	O
the	O
WT	B-protein
hIL-4Ralpha	I-protein
in	O
driving	O
Th2	O
differentiation	O
,	O
as	O
measured	O
by	O
Gata3	O
up-regulation	O
and	O
IL-4	O
production	O
.	O

Furthermore	O
,	O
IL-4-driven	O
cell	O
expansion	O
was	O
also	O
normal	O
in	O
the	O
cells	O
infected	O
with	O
Y497F	B-protein
,	O
although	O
cells	O
infected	O
with	O
Y497F	B-protein
were	O
not	O
capable	O
of	O
phosphorylating	O
insulin	B-protein
receptor	I-protein
substrate	I-protein
2	I-protein
.	O

These	O
results	O
suggest	O
that	O
the	O
signal	O
pathway	O
mediated	O
by	O
Y497	B-protein
is	O
dispensable	O
for	O
both	O
IL-4	B-protein
-driven	O
Th2	O
differentiation	O
and	O
cell	O
expansion	O
.	O

Both	O
WT	B-protein
and	I-protein
Y497F	I-protein
hIL-4Ralpha	I-protein
lose	O
the	O
ability	O
to	O
drive	O
Th2	O
differentiation	O
and	O
cell	O
expansion	O
in	O
Stat6-knockout	B-cell_type
CD4	I-cell_type
T	I-cell_type
cells	I-cell_type
.	O

A	O
constitutively	O
activated	O
form	O
of	O
Stat6	B-protein
introduced	O
into	O
CD4	B-cell_type
T	I-cell_type
cells	I-cell_type
resulted	O
in	O
both	O
Th2	O
differentiation	O
and	O
enhanced	O
cell	O
expansion	O
.	O

Thus	O
,	O
activated	B-protein
Stat6	I-protein
is	O
necessary	O
and	O
sufficient	O
to	O
mediate	O
both	O
IL-4	B-protein
-driven	O
Th2	O
differentiation	O
and	O
cell	O
expansion	O
in	O
CD4	B-cell_type
T	I-cell_type
cells	I-cell_type
.	O

-DOCSTART-	O

The	O
effect	O
of	O
HIV-1	B-protein
regulatory	I-protein
proteins	I-protein
on	O
cellular	O
genes	O
:	O
derepression	O
of	O
the	O
IL-2	B-DNA
promoter	I-DNA
by	O
Tat	B-protein
.	O

In	O
HIV-infected	O
individuals	O
dysregulation	O
of	O
the	O
immune	O
system	O
is	O
characterized	O
by	O
severe	O
disorders	O
of	O
the	O
cytokine	O
network	O
.	O

Increase	O
secretion	O
of	O
IL-2	B-protein
,	O
the	O
major	O
T	B-cell_type
cell	I-cell_type
growth	O
and	O
differentiation	O
cytokine	B-protein
,	O
may	O
play	O
a	O
decisive	O
role	O
in	O
sensitization	O
of	O
T	B-cell_type
cells	I-cell_type
for	O
activation	O
induced	O
apoptosis	O
and	O
indirect	O
death	O
of	O
activated	B-cell_type
T	I-cell_type
cells	I-cell_type
through	O
augmented	O
virus	O
replication	O
.	O

We	O
investigated	O
the	O
cause	O
of	O
enhanced	O
IL-2	O
secretion	O
and	O
found	O
that	O
the	O
HIV	B-protein
Tat	I-protein
induces	O
this	O
effect	O
.	O

We	O
demonstrate	O
that	O
increased	O
IL-2	O
secretion	O
is	O
due	O
to	O
Tat	B-protein
-enhanced	O
IL-2	B-DNA
promoter	I-DNA
activation	O
.	O

Tat	B-protein
derepresses	O
and	O
activates	O
the	O
distal	B-DNA
AP-1	I-DNA
site	I-DNA
(	O
position	B-DNA
-185	I-DNA
to	I-DNA
-177	I-DNA
)	O
in	O
the	O
IL-2	B-DNA
promoter	I-DNA
.	O

In	O
nonstimulated	B-cell_type
T	I-cell_type
cells	I-cell_type
a	O
repressor	O
complex	O
containing	O
NF-IL6	B-protein
,	O
JunB	B-protein
,	O
c-Fos	B-protein
and	O
Fra-1	B-protein
is	O
formed	O
on	O
the	O
AP-1	B-DNA
(	I-DNA
IL-2/d	I-DNA
)	I-DNA
site	I-DNA
and	O
represses	O
IL-2	B-DNA
promoter	I-DNA
activity	O
.	O

After	O
T	O
cell	O
activation	O
,	O
a	O
heterodimeric	B-protein
activator	I-protein
containing	O
p65	B-protein
and	O
c-Jun	B-protein
binds	O
to	O
the	O
AP-1	B-DNA
(	I-DNA
IL-2/d	I-DNA
)	I-DNA
site	I-DNA
.	O

HIV	B-protein
Tat	I-protein
enhances	O
activation	O
of	O
NF-kappaB	B-protein
and	O
consequently	O
,	O
activates	O
the	O
AP-1	B-DNA
(	I-DNA
IL-2/d	I-DNA
)	I-DNA
site	I-DNA
.	O

Our	O
data	O
provide	O
evidence	O
for	O
a	O
novel	O
mechanism	O
by	O
which	O
HIV	B-protein
Tat	I-protein
dysregulates	O
IL-2	O
production	O
and	O
therefore	O
may	O
contribute	O
to	O
the	O
HIV-1	O
infection	O
in	O
a	O
way	O
yet	O
to	O
be	O
clarified	O
.	O

-DOCSTART-	O

Transforming	B-protein
growth	I-protein
factor-beta1	I-protein
interferes	O
with	O
thrombopoietin-induced	O
signal	O
transduction	O
in	O
megakaryoblastic	B-cell_type
and	I-cell_type
erythroleukemic	I-cell_type
cells	I-cell_type
.	O

OBJECTIVE	O
:	O
Thrombopoietin	B-protein
(	O
TPO	B-protein
)	O
and	O
transforming	B-protein
growth	I-protein
factor-beta	I-protein
(	I-protein
1	I-protein
)	I-protein
(	O
TGF-beta	B-protein
(	I-protein
1	I-protein
)	I-protein
)	O
have	O
been	O
shown	O
to	O
exert	O
opposite	O
effects	O
on	O
proliferation	O
and	O
megakaryocytic	O
differentiation	O
of	O
hematopoietic	B-cell_type
cells	I-cell_type
.	O

To	O
determine	O
whether	O
TGF-beta	B-protein
(	I-protein
1	I-protein
)	I-protein
interferes	O
directly	O
with	O
TPO	B-protein
-induced	O
signal	O
transduction	O
in	O
hematopoietic	B-cell_type
cells	I-cell_type
,	O
we	O
compared	O
the	O
regulatory	O
effects	O
in	O
the	O
TPO	B-protein
-responsive	O
cell	O
lines	O
Mo-7e	O
and	O
HEL	O
.	O

MATERIALS	O
AND	O
METHODS	O
:	O
The	O
cells	O
were	O
stimulated	O
by	O
100	O
ng/mL	O
TPO	B-protein
and/or	O
100	O
ng/mL	O
TGF-beta1	B-protein
and	O
analyzed	O
for	O
proliferation	O
(	O
3H	O
thymidine	O
incorporation	O
)	O
,	O
viability	O
(	O
trypan	O
blue	O
exclusion	O
)	O
,	O
and	O
protein	O
expression	O
and	O
phosphorylation	O
(	O
Western	O
blot	O
)	O
.	O

RESULTS	O
:	O
TPO	B-protein
enhanced	O
the	O
proliferation	O
of	O
Mo-7e	B-cell_line
cells	I-cell_line
as	O
determined	O
by	O
3H-thymidine	O
incorporation	O
,	O
whereas	O
TGF-beta1	B-protein
suppressed	O
baseline	O
cell	O
growth	O
and	O
antagonized	O
the	O
proliferative	O
effect	O
of	O
TPO	B-protein
.	O

TPO	B-protein
-induced	O
proliferation	O
also	O
was	O
reduced	O
by	O
a	O
specific	O
inhibitor	O
of	O
the	O
mitogen-activated	B-protein
protein	I-protein
kinase	I-protein
(	O
MAPK	B-protein
)	O
pathway	O
(	O
PD098059	O
)	O
,	O
which	O
inhibits	O
activation	O
of	O
the	O
MAPK	B-protein
extracellular	I-protein
signal-regulated	I-protein
kinases	I-protein
(	O
ERK	B-protein
)	O
ERK1	B-protein
and	O
ERK2	B-protein
,	O
and	O
AG490	O
,	O
an	O
inhibitor	O
of	O
Janus	B-protein
kinase-2	I-protein
,	O
which	O
completely	O
blocked	O
TPO	B-protein
-induced	O
proliferation	O
.	O

As	O
demonstrated	O
by	O
Western	O
blotting	O
,	O
TGF-beta1	B-protein
reduced	O
the	O
TPO	B-protein
-stimulated	O
ERK1	B-protein
/	O
ERK2	B-protein
and	O
STAT5	O
phosphorylation	O
in	O
Mo-7e	B-cell_line
and	I-cell_line
HEL	I-cell_line
cells	I-cell_line
.	O

This	O
effect	O
was	O
completely	O
reversed	O
by	O
preincubation	O
with	O
a	O
tyrosine	O
phosphatase	O
inhibitor	O
(	O
Na3VO4	O
)	O
,	O
which	O
suggests	O
that	O
TGF-beta1	B-protein
activated	O
a	O
phosphatase	B-protein
.	O

Although	O
STAT3	B-protein
also	O
was	O
activated	O
by	O
TPO	B-protein
,	O
STAT3	B-protein
activation	O
remained	O
unaltered	O
by	O
TGF-beta1	B-protein
.	O

CONCLUSION	O
:	O
Taken	O
together	O
,	O
these	O
data	O
suggest	O
that	O
TGF-beta1	B-protein
modulates	O
TPO	B-protein
-mediated	O
effects	O
on	O
megakaryocytic	O
proliferation	O
by	O
interfering	O
with	O
TPO	B-protein
-induced	O
signal	O
transduction	O
,	O
particularly	O
by	O
reducing	O
the	O
activities	O
of	O
MAPK	B-protein
ERK1/ERK2	I-protein
and	I-protein
STAT5	I-protein
.	O

-DOCSTART-	O

Invariant	B-protein
chain	I-protein
induces	O
B	O
cell	O
maturation	O
by	O
activating	O
a	O
TAF	B-protein
(	I-protein
II	I-protein
)	I-protein
105-NF-kappaB	I-protein
-dependent	O
transcription	O
program	O
.	O

Early	O
stages	O
of	O
B	O
cell	O
development	O
occur	O
in	O
the	O
bone	O
marrow	O
,	O
resulting	O
in	O
formation	O
of	O
immature	B-cell_type
B	I-cell_type
cells	I-cell_type
.	O

From	O
there	O
these	O
immature	B-cell_type
cells	I-cell_type
migrate	O
to	O
the	O
spleen	O
where	O
they	O
differentiate	O
to	O
mature	B-cell_type
cells	I-cell_type
.	O

This	O
final	O
maturation	O
step	O
is	O
crucial	O
for	O
the	O
B	B-cell_type
cells	I-cell_type
to	O
become	O
responsive	O
to	O
antigens	B-protein
and	O
to	O
participate	O
in	O
the	O
immune	O
response	O
.	O

Recently	O
,	O
invariant	B-protein
chain	I-protein
(	O
Ii	B-protein
)	O
,	O
a	O
major	B-protein
histocompatibility	I-protein
complex	I-protein
class	I-protein
II	I-protein
chaperone	I-protein
,	O
as	O
well	O
as	O
the	O
transcription	B-protein
factors	I-protein
c-Rel	B-protein
and	O
p65/RelA	B-protein
,	O
were	O
found	O
to	O
play	O
a	O
role	O
in	O
the	O
final	O
antigen-independent	O
differentiation	O
stage	O
of	O
B	B-cell_type
cells	I-cell_type
in	O
the	O
spleen	O
.	O

In	O
this	O
study	O
,	O
we	O
investigated	O
a	O
possible	O
link	O
between	O
Ii	B-protein
-dependent	O
B	O
cell	O
maturation	O
and	O
the	O
NF-kappaB	O
pathway	O
.	O

Our	O
studies	O
indicate	O
that	O
Ii	B-protein
-induced	O
B	O
cell	O
maturation	O
involves	O
activation	O
of	O
transcription	O
mediated	O
by	O
the	O
NF-kappaB	B-protein
p65/RelA	I-protein
homodimer	I-protein
and	O
requires	O
the	O
B	B-protein
cell-enriched	I-protein
coactivator	I-protein
TBP-associated	B-protein
factor	I-protein
(	I-protein
II	I-protein
)	I-protein
105	I-protein
.	O

-DOCSTART-	O

Androgens	O
indirectly	O
accelerate	O
thymocyte	B-cell_type
apoptosis	O
.	O

Apoptotic	O
processes	O
,	O
or	O
the	O
disturbance	O
of	O
the	O
natural	O
regulation	O
of	O
these	O
processes	O
,	O
may	O
be	O
involved	O
in	O
the	O
pathogenesis	O
of	O
autoimmune	O
diseases	O
(	O
AID	O
)	O
.	O

Women	O
are	O
,	O
in	O
general	O
,	O
more	O
susceptible	O
than	O
men	O
to	O
develop	O
AID	O
like	O
rheumatoid	O
arthritis	O
.	O

Androgens	O
and	O
glucocorticoids	O
,	O
in	O
contrast	O
to	O
oestrogens	O
,	O
have	O
favourable	O
effects	O
in	O
AID	O
models	O
as	O
well	O
as	O
in	O
human	O
AID	O
.	O

It	O
is	O
known	O
that	O
glucocorticoids	O
(	O
GC	O
)	O
,	O
used	O
for	O
treatment	O
of	O
AID	O
,	O
increase	O
apoptosis	O
in	O
the	O
thymus	O
resulting	O
in	O
decreased	O
numbers	O
of	O
CD4+	B-cell_type
CD8+	I-cell_type
thymocytes	I-cell_type
.	O

It	O
was	O
asked	O
whether	O
androgens	O
,	O
in	O
contrast	O
to	O
oestrogens	O
,	O
exert	O
their	O
favourable	O
effects	O
in	O
the	O
treatment	O
of	O
AID	O
by	O
a	O
mechanism	O
comparable	O
to	O
that	O
described	O
for	O
GC	O
by	O
eliminating	O
the	O
apoptosis	O
prone	O
CD4+	B-cell_type
CD8+	I-cell_type
population	I-cell_type
in	O
the	O
thymus	O
.	O

Although	O
both	O
androgens	O
and	O
oestrogens	O
proved	O
thymolytic	O
,	O
a	O
significantly	O
decreased	O
percentage	O
of	O
CD4+	B-cell_type
CD8+	I-cell_type
thymocytes	I-cell_type
was	O
observed	O
by	O
flow	O
cytometry	O
after	O
treatment	O
of	O
mice	O
with	O
the	O
androgen	O
methyltestosterone	O
,	O
but	O
not	O
with	O
the	O
oestrogen	O
ethinylestradiol	O
.	O

To	O
investigate	O
whether	O
the	O
observed	O
thymolytic	O
effects	O
were	O
due	O
to	O
the	O
presence	O
of	O
hormone	B-protein
receptors	I-protein
on	O
thymocytes	B-cell_type
,	O
cells	O
were	O
isolated	O
from	O
the	O
thymus	O
and	O
incubated	O
with	O
androgens	O
or	O
oestrogens	O
to	O
measure	O
apoptosis	O
.	O

Several	O
techniques	O
were	O
used	O
to	O
determine	O
thymocyte	B-cell_type
apoptosis	O
in	O
vitro	O
,	O
but	O
no	O
enhanced	O
apoptotic	O
signal	O
was	O
observed	O
.	O

Using	O
the	O
very	O
sensitive	O
TUNEL	O
assay	O
,	O
no	O
direct	O
effect	O
of	O
androgens	O
on	O
thymocytes	B-cell_type
in	O
vitro	O
could	O
be	O
observed	O
.	O

This	O
is	O
in	O
sharp	O
contrast	O
to	O
the	O
high	O
signal	O
observed	O
with	O
GC	O
.	O

Therefore	O
,	O
upon	O
in	O
vivo	O
androgen	O
treatment	O
,	O
other	O
cells	O
containing	O
androgen	B-protein
receptors	I-protein
than	O
thymocytes	B-cell_type
are	O
probably	O
involved	O
in	O
inducing	O
the	O
increase	O
in	O
thymic	O
apoptosis	O
.	O

To	O
study	O
the	O
role	O
of	O
the	O
androgen	B-protein
receptor	I-protein
on	O
thymocyte	B-cell_type
apoptosis	O
,	O
androgen	B-protein
receptor	I-protein
mutant	O
(	O
Tfm/Y	O
)	O
mice	O
were	O
treated	O
with	O
androgens	O
.	O

No	O
alterations	O
of	O
thymocyte	B-cell_type
subpopulations	I-cell_type
were	O
seen	O
,	O
suggesting	O
that	O
changes	O
in	O
the	O
percentage	O
of	O
CD4+	B-cell_type
CD8+	I-cell_type
thymocytes	I-cell_type
after	O
administration	O
of	O
androgens	O
depend	O
on	O
the	O
presence	O
of	O
functional	O
androgen	B-protein
receptors	I-protein
.	O

Thus	O
,	O
it	O
is	O
concluded	O
that	O
androgens	O
indirectly	O
accelerate	O
thymocyte	B-cell_type
apoptosis	O
in	O
vivo	O
.	O

-DOCSTART-	O

Inhibition	O
of	O
the	O
transcription	B-protein
factors	I-protein
AP-1	B-protein
and	O
NF-kappaB	B-protein
in	O
CD4	B-cell_type
T	I-cell_type
cells	I-cell_type
by	O
peroxisome	B-protein
proliferator-activated	I-protein
receptor	I-protein
gamma	I-protein
ligands	I-protein
.	O

The	O
peroxisome	B-protein
proliferator-activated	I-protein
receptor	I-protein
gamma	I-protein
(	O
PPARgamma	B-protein
)	O
,	O
a	O
member	O
of	O
the	O
nuclear	B-protein
hormone	I-protein
receptor	I-protein
superfamily	I-protein
,	O
is	O
essential	O
for	O
adipocyte	O
differentiation	O
and	O
glucose	O
homeostasis	O
.	O

PPARgamma	B-protein
has	O
been	O
found	O
recently	O
to	O
regulate	O
macrophage	O
activation	O
in	O
response	O
to	O
mitogens	B-protein
and	O
inflammation	O
.	O

Our	O
study	O
shows	O
PPARgamma	B-protein
to	O
be	O
preferentially	O
expressed	O
in	O
the	O
nuclei	O
of	O
resting	B-cell_type
T	I-cell_type
cells	I-cell_type
and	O
to	O
increase	O
upon	O
activation	O
of	O
T	B-cell_type
cells	I-cell_type
by	O
either	O
anti-CD3	B-protein
and	O
anti-CD28	B-protein
or	O
phorbol	O
myristyl	O
acetate	O
(	O
PMA	O
)	O
.	O

We	O
also	O
found	O
the	O
PPARgamma	B-protein
ligand	I-protein
ciglitizone	O
to	O
attenuate	O
the	O
activation	O
of	O
T	B-cell_type
cells	I-cell_type
by	O
inhibiting	O
cytokine	O
gene	O
expression	O
and	O
anti-CD3	B-protein
and	O
anti-CD28	B-protein
or	O
PMA-induced	O
proliferative	O
responses	O
.	O

Inhibition	O
of	O
both	O
the	O
proliferative	O
response	O
and	O
inflammatory	O
cytokine	O
expression	O
in	O
CD4	B-cell_type
T	I-cell_type
cells	I-cell_type
was	O
correlated	O
with	O
suppression	O
of	O
the	O
activated	O
transcription	B-protein
factors	I-protein
AP1	B-protein
and	O
NF-kappaB	B-protein
.	O

PPARgamma	B-protein
ligands	I-protein
also	O
strongly	O
inhibited	O
SEA-induced	O
Vbeta3	O
T	O
cell	O
activation	O
in	O
vivo	O
.	O

These	O
results	O
,	O
together	O
with	O
previous	O
findings	O
of	O
the	O
inhibitory	O
effect	O
of	O
PPARgamma	B-protein
ligands	I-protein
on	O
activated	B-cell_type
macrophages	I-cell_type
,	O
provide	O
clear	O
evidence	O
for	O
PPARgamma	B-protein
as	O
a	O
negative	B-protein
regulator	I-protein
of	O
the	O
inflammatory	O
activation	O
of	O
both	O
macrophage	B-cell_type
and	O
T	B-cell_type
cells	I-cell_type
.	O

PPARgamma	B-protein
may	O
thus	O
be	O
a	O
potential	O
therapeutic	O
target	O
for	O
the	O
treatment	O
of	O
autoimmunity	O
.	O

-DOCSTART-	O

A	O
prominent	O
role	O
for	O
activator	B-protein
protein-1	I-protein
in	O
the	O
transcription	O
of	O
the	O
human	B-DNA
2B4	I-DNA
(	I-DNA
CD244	I-DNA
)	I-DNA
gene	I-DNA
in	O
NK	B-cell_type
cells	I-cell_type
.	O

The	O
cell	B-protein
surface	I-protein
glycoprotein	I-protein
2B4	I-protein
(	O
CD244	B-protein
)	O
of	O
the	O
Ig	B-protein
superfamily	I-protein
is	O
involved	O
in	O
the	O
regulation	O
of	O
NK	B-cell_type
and	O
T	B-cell_type
lymphocyte	I-cell_type
functions	O
.	O

We	O
have	O
recently	O
identified	O
CD48	B-protein
as	O
the	O
high	B-protein
affinity	I-protein
counterreceptor	I-protein
for	O
2B4	B-protein
in	O
both	O
mice	O
and	O
humans	O
.	O

The	O
cytoplasmic	B-protein
domain	I-protein
of	O
2B4	B-protein
associates	O
with	O
src	B-protein
homology	I-protein
2	I-protein
domain-containing	I-protein
protein	I-protein
or	O
signaling	B-protein
lymphocyte	I-protein
activation	I-protein
molecule-associated	I-protein
protein	I-protein
,	O
whose	O
mutation	O
is	O
the	O
underlying	O
genetic	O
defect	O
in	O
the	O
X-linked	O
lymphoproliferative	O
syndrome	O
.	O

In	O
this	O
study	O
,	O
we	O
report	O
the	O
molecular	O
cloning	O
and	O
characterization	O
of	O
the	O
human	B-DNA
2B4	I-DNA
(	I-DNA
h2B4	I-DNA
)	I-DNA
promoter	I-DNA
.	O

Through	O
primer	O
extension	O
analysis	O
,	O
we	O
found	O
that	O
the	O
transcription	O
of	O
the	O
h2B4	B-DNA
gene	I-DNA
initiates	O
at	O
multiple	O
start	O
sites	O
.	O

We	O
isolated	O
h2B4	B-DNA
genomic	I-DNA
clones	I-DNA
and	O
PCR	O
amplified	O
the	O
5	B-DNA
'	I-DNA
untranslated	I-DNA
region	I-DNA
containing	O
the	O
promoter	B-DNA
elements	I-DNA
.	O

We	O
have	O
identified	O
a	O
functional	O
AP-1	B-DNA
site	I-DNA
that	O
lies	O
between	O
(	O
-106	O
to	O
-100	O
)	O
through	O
transient	O
transfection	O
analysis	O
in	O
YT	B-cell_line
cells	I-cell_line
,	O
a	O
human	B-cell_line
NK	I-cell_line
cell	I-cell_line
line	I-cell_line
.	O

EMSAs	O
with	O
Abs	B-protein
specific	O
for	O
various	O
protein	B-protein
factors	I-protein
of	O
the	O
AP-1	B-protein
family	I-protein
revealed	O
that	O
multiple	O
members	O
of	O
the	O
Jun	B-protein
family	I-protein
are	O
involved	O
in	O
the	O
regulation	O
of	O
the	O
h2B4	B-DNA
gene	I-DNA
.	O

Mutation	O
of	O
the	O
AP-1	B-DNA
site	I-DNA
not	O
only	O
abolishes	O
protein/DNA	O
interactions	O
but	O
also	O
promoter	O
activity	O
.	O

These	O
results	O
demonstrate	O
a	O
significant	O
role	O
for	O
AP-1	B-protein
in	O
the	O
transcriptional	O
regulation	O
of	O
the	O
h2B4	B-DNA
gene	I-DNA
.	O

-DOCSTART-	O

Functional	O
correction	O
of	O
FA-C	B-cell_type
cells	I-cell_type
with	O
FANCC	B-protein
suppresses	O
the	O
expression	O
of	O
interferon	B-DNA
gamma-inducible	I-DNA
genes	I-DNA
.	O

Because	O
hematopoietic	B-cell_type
cells	I-cell_type
derived	O
from	O
Fanconi	O
anemia	O
(	O
FA	O
)	O
patients	O
of	O
the	O
C-complementation	O
group	O
(	O
FA-C	O
)	O
are	O
hypersensitive	O
to	O
the	O
inhibitory	O
effects	O
of	O
interferon	B-protein
gamma	I-protein
(	O
IFNgamma	B-protein
)	O
,	O
the	O
products	O
of	O
certain	O
IFNgamma-inducible	B-DNA
genes	I-DNA
known	O
to	O
influence	O
hematopoietic	O
cell	O
survival	O
were	O
quantified	O
.	O

High	O
constitutive	O
expression	O
of	O
the	O
IFNgamma-inducible	B-DNA
genes	I-DNA
,	O
IFN-stimulated	B-protein
gene	I-protein
factor	I-protein
3	I-protein
gamma	I-protein
subunit	I-protein
(	O
ISGF3gamma	B-protein
)	O
,	O
IFN	B-protein
regulatory	I-protein
factor-1	I-protein
(	O
IRF-1	B-protein
)	O
,	O
and	O
the	O
cyclin-dependent	B-protein
kinase	I-protein
inhibitor	I-protein
p21	I-protein
(	O
WAF1	B-protein
)	O
was	O
found	O
in	O
FANCC	B-protein
mutant	O
B	B-cell_type
lymphoblasts	I-cell_type
,	O
low-density	B-cell_type
bone	I-cell_type
marrow	I-cell_type
cells	I-cell_type
,	O
and	O
murine	B-cell_type
embryonic	I-cell_type
fibroblasts	I-cell_type
.	O

Paradoxically	O
,	O
these	O
cells	O
do	O
not	O
activate	B-protein
signal	I-protein
transducer	I-protein
and	I-protein
activator	I-protein
of	I-protein
transcription	I-protein
(	I-protein
STAT	I-protein
)	I-protein
1	I-protein
properly	O
.	O

In	O
an	O
attempt	O
to	O
clarify	O
mechanisms	O
by	O
which	O
FA-C	B-cell_type
cells	I-cell_type
overexpress	O
IFNgamma-inducible	B-DNA
genes	I-DNA
in	O
the	O
face	O
of	O
defective	O
STAT1	O
phosphorylation	O
,	O
it	O
was	O
reasoned	O
that	O
decreased	O
levels	O
of	O
activated	B-protein
STAT1	I-protein
might	O
result	O
in	O
reduced	O
expression	O
of	O
a	O
hematopoietic	B-protein
IFNgamma-responsive	I-protein
protein	I-protein
that	O
normally	O
modulates	O
expression	O
of	O
other	O
IFNgamma-responsive	B-DNA
genes	I-DNA
.	O

Levels	O
of	O
the	O
IFNgamma	B-protein
-inducible	O
factor	O
IFN	B-protein
consensus	I-protein
sequence	I-protein
binding	I-protein
protein	I-protein
(	O
ICSBP	B-protein
)	O
,	O
a	O
negative	B-protein
trans-acting	I-protein
regulator	I-protein
of	O
some	O
IFNgamma-inducible	B-DNA
genes	I-DNA
,	O
were	O
quantified	O
.	O

ICSBP	B-protein
levels	O
were	O
reduced	O
in	O
FA-C	B-cell_type
B	I-cell_type
lymphoblasts	I-cell_type
and	O
MEFs	B-cell_type
.	O

However	O
,	O
enforced	O
expression	O
of	O
ICSBP	B-protein
failed	O
to	O
down-regulate	O
IRF-1	B-protein
,	O
ISGF3gamma	B-protein
,	O
and	O
p21	B-protein
(	I-protein
WAF1	I-protein
)	I-protein
.	O

Thus	O
,	O
the	O
FANCC	B-protein
protein	I-protein
functions	O
to	O
modulate	O
expression	O
of	O
a	O
family	O
of	O
genes	O
that	O
in	O
normal	O
cells	O
are	O
inducible	O
only	O
by	O
specific	O
environmental	O
cues	O
for	O
apoptosis	O
or	O
mitogenic	O
inhibition	O
,	O
but	O
it	O
does	O
so	O
independently	O
of	O
the	O
classic	O
IFN-STAT1	O
pathway	O
and	O
is	O
not	O
the	O
direct	O
result	O
of	O
reduced	O
ICSBP	B-protein
expression	O
.	O

-DOCSTART-	O

A	O
genetic	O
investigation	O
of	O
E2A	O
function	O
in	O
lymphocyte	O
development	O
.	O

Lymphocytes	B-cell_type
are	O
derived	O
from	O
hematopoietic	B-cell_type
stem	I-cell_type
cells	I-cell_type
(	O
HSC	B-cell_type
)	O
following	O
a	O
series	O
of	O
regulated	O
differentiation	O
events	O
.	O

Multipotent	B-cell_type
HSCs	I-cell_type
become	O
committed	O
to	O
the	O
B	B-cell_line
cell	I-cell_line
lineage	I-cell_line
in	O
bone	O
marrow	O
and	O
the	O
T	B-cell_line
cell	I-cell_line
lineage	I-cell_line
in	O
the	O
thymus	O
after	O
receiving	O
appropriate	O
signals	O
from	O
the	O
corresponding	O
microenvironment	O
.	O

These	O
committed	O
lymphoid	B-cell_type
cells	I-cell_type
must	O
then	O
undergo	O
V	O
(	O
D	O
)	O
J	O
recombination	O
at	O
the	O
immunoglobulin	B-DNA
gene	I-DNA
or	O
T	B-DNA
cell	I-DNA
receptor	I-DNA
gene	I-DNA
locus	I-DNA
resulting	O
in	O
clonal	O
production	O
of	O
functional	O
B	B-cell_type
or	I-cell_type
T	I-cell_type
lymphocytes	I-cell_type
,	O
respectively	O
.	O

Lymphocyte	O
commitment	O
and	O
differentiation	O
are	O
accompanied	O
by	O
programmed	O
gene	O
expression	O
or	O
repression	O
events	O
which	O
are	O
driven	O
by	O
lineage	B-protein
and	I-protein
stage	I-protein
specific	I-protein
transcription	I-protein
factors	I-protein
.	O

The	O
basic-helix-loop-helix	B-protein
(	I-protein
bHLH	I-protein
)	I-protein
transcription	I-protein
factors	I-protein
encoded	O
by	O
the	O
E2A	B-DNA
gene	I-DNA
are	O
involved	O
in	O
several	O
differentiation	O
events	O
during	O
B	O
and	O
T	O
cell	O
development	O
,	O
including	O
lineage	O
commitment	O
,	O
initiation	O
of	O
V	O
(	O
D	O
)	O
J	O
recombination	O
,	O
and	O
antigen	O
receptor	O
mediated	O
proliferation	O
and	O
differentiation	O
.	O

Several	O
recent	O
reviews	O
have	O
provided	O
a	O
comprehensive	O
discussion	O
of	O
biochemical	O
,	O
cellular	O
,	O
and	O
genetic	O
research	O
on	O
E2A	O
function	O
in	O
lymphocyte	O
development	O
(	O
1	O
,	O
2	O
)	O
.	O

Here	O
,	O
we	O
only	O
discuss	O
some	O
of	O
the	O
genetic	O
approaches	O
our	O
laboratory	O
(	O
except	O
where	O
it	O
is	O
noted	O
)	O
has	O
undertaken	O
to	O
investigate	O
the	O
molecular	O
pathways	O
mediated	O
by	O
E2A	B-protein
transcription	I-protein
factors	I-protein
in	O
lymphocyte	O
development	O

-DOCSTART-	O

D609	O
inhibits	O
ionizing	O
radiation-induced	O
oxidative	O
damage	O
by	O
acting	O
as	O
a	O
potent	O
antioxidant	O
.	O

Tricyclodecan-9-yl-xanthogenate	O
(	O
D609	O
)	O
has	O
been	O
extensively	O
studied	O
in	O
biological	O
systems	O
and	O
exhibits	O
a	O
variety	O
of	O
biological	O
functions	O
,	O
including	O
antiviral	O
,	O
antitumor	O
,	O
and	O
anti-inflammatory	O
activities	O
.	O

Most	O
of	O
these	O
activities	O
have	O
been	O
largely	O
attributed	O
to	O
the	O
inhibitory	O
effect	O
of	O
D609	O
on	O
phosphatidylcholine-specific	O
phospholipase	B-protein
C	I-protein
.	O

However	O
,	O
as	O
a	O
xanthate	O
derivative	O
,	O
D609	O
is	O
a	O
strong	O
electrolyte	O
and	O
readily	O
dissociates	O
to	O
xanthate	O
anions	O
and	O
cations	O
of	O
alkali	O
metals	O
in	O
solution	O
.	O

Xanthate	O
anions	O
and	O
protonated	O
xanthic	O
acid	O
contain	O
a	O
free	O
thiol	O
moiety	O
and	O
are	O
highly	O
reductive	O
.	O

This	O
implies	O
that	O
D609	O
and	O
other	O
xanthate	O
derivatives	O
may	O
function	O
as	O
potent	O
antioxidants	O
.	O

Indeed	O
,	O
we	O
found	O
that	O
D609	O
inhibited	O
the	O
Fenton	O
reaction-induced	O
oxidation	O
of	O
dihydrorhodamine	O
123	O
in	O
a	O
dose-dependent	O
manner	O
similar	O
to	O
that	O
of	O
pyrrolidinedithiocarbamate	O
,	O
a	O
well	O
known	O
antioxidant	O
.	O

In	O
addition	O
,	O
D609	O
inhibited	O
the	O
formation	O
of	O
the	O
alpha-phenyl-tert-butylnitrone-free	O
radical	O
spin	O
adducts	O
and	O
lipid	O
peroxidation	O
of	O
synaptosomal	O
membranes	O
by	O
the	O
Fenton	O
reagents	O
.	O

Furthermore	O
,	O
preincubation	O
of	O
lymphocytes	B-cell_type
with	O
D609	O
resulted	O
in	O
a	O
significant	O
diminution	O
of	O
ionizing	O
radiation	O
(	O
IR	O
)	O
-induced	O
1	O
)	O
production	O
of	O
reactive	O
oxygen	O
species	O
;	O
2	O
)	O
decrease	O
in	O
intracellular	O
reduced	O
glutathione	O
;	O
3	O
)	O
oxidative	O
damage	O
to	O
proteins	O
and	O
lipids	O
;	O
and	O
4	O
)	O
activation	O
of	O
nuclear	B-protein
factor-kappaB	I-protein
.	O

Moreover	O
,	O
when	O
D609	O
(	O
50	O
mg/kg	O
i.v.	O
)	O
was	O
administered	O
to	O
mice	O
10	O
min	O
prior	O
to	O
total	O
body	O
IR	O
(	O
6.5	O
and	O
8.5	O
Gy	O
)	O
,	O
it	O
protected	O
the	O
mice	O
from	O
IR-induced	O
lethality	O
.	O

Thus	O
,	O
these	O
results	O
indicate	O
that	O
D609	O
is	O
a	O
potent	O
antioxidant	O
and	O
has	O
the	O
ability	O
to	O
inhibit	O
IR-induced	O
cellular	O
oxidative	O
stress	O
.	O

-DOCSTART-	O

Inhibition	O
of	O
AP-1	B-protein
by	O
the	O
glucocorticoid-inducible	O
protein	B-protein
GILZ	I-protein
.	O

The	O
immunosuppressive	O
effects	O
of	O
glucocorticoids	O
arise	O
largely	O
by	O
inhibition	O
of	O
cytokine	O
gene	O
expression	O
,	O
which	O
has	O
been	O
ascribed	O
to	O
interference	O
between	O
the	O
glucocorticoid	B-protein
receptor	I-protein
and	O
transcription	B-protein
factors	I-protein
such	O
as	O
AP-1	B-protein
and	O
NF-kappa	B-protein
B	I-protein
as	O
well	O
as	O
by	O
competition	O
for	O
common	O
coactivators	O
.	O

Here	O
we	O
show	O
that	O
glucocorticoid-induced	O
inhibition	O
of	O
interleukin-2	O
mRNA	O
expression	O
in	O
activated	B-cell_type
normal	I-cell_type
T	I-cell_type
cells	I-cell_type
required	O
new	O
protein	O
synthesis	O
,	O
suggesting	O
that	O
this	O
phenomenon	O
is	O
secondary	O
to	O
expression	O
of	O
glucocorticoid-regulated	O
genes	B-DNA
.	O

One	O
of	O
the	O
most	O
prominent	O
glucocorticoid-induced	O
genes	B-DNA
is	O
glucocorticoid-induced	B-DNA
leucine	I-DNA
zipper	I-DNA
(	O
GILZ	B-DNA
)	O
,	O
which	O
has	O
been	O
reported	O
to	O
inhibit	O
activation-induced	O
up-regulation	O
of	O
Fas	B-RNA
ligand	I-RNA
(	I-RNA
FasL	I-RNA
)	I-RNA
mRNA	I-RNA
.	O

Indeed	O
,	O
transient	O
expression	O
of	O
GILZ	B-DNA
in	O
Jurkat	B-cell_line
T	I-cell_line
cells	I-cell_line
blocked	O
induction	O
of	O
a	O
reporter	B-DNA
construct	I-DNA
driven	O
by	O
the	O
FasL	B-DNA
promoter	I-DNA
.	O

This	O
could	O
be	O
accounted	O
for	O
by	O
GILZ	B-protein
-mediated	O
inhibition	O
of	O
Egr-2	B-protein
and	O
Egr-3	B-protein
,	O
NFAT/AP-1	B-protein
-inducible	O
transcription	B-protein
factors	I-protein
that	O
bind	O
a	O
regulatory	B-DNA
element	I-DNA
in	O
the	O
FasL	B-DNA
promoter	I-DNA
and	O
up-regulate	O
FasL	O
expression	O
.	O

GILZ	B-protein
also	O
potently	O
inhibited	O
AP-1-driven	B-DNA
and	I-DNA
IL-2	I-DNA
promoter-driven	I-DNA
reporter	I-DNA
constructs	I-DNA
,	O
and	O
recombinant	O
GILZ	B-protein
specifically	O
interacted	O
with	O
c-Fos	B-protein
and	O
c-Jun	B-protein
in	O
vitro	O
and	O
inhibited	O
the	O
binding	O
of	O
active	B-protein
AP-1	I-protein
to	O
its	O
target	O
DNA	O
.	O

Whereas	O
homodimerization	O
of	O
GILZ	B-protein
required	O
the	O
presence	O
of	O
its	O
leucine	B-protein
zipper	I-protein
,	O
the	O
interaction	O
with	O
c-Fos	B-protein
and	O
c-Jun	B-protein
occurred	O
through	O
the	O
N-terminal	B-protein
60-amino	B-protein
acid	I-protein
region	I-protein
of	O
GILZ	B-protein
.	O

Thus	O
,	O
GILZ	B-protein
represents	O
a	O
glucocorticoid-induced	O
gene	O
product	O
that	O
can	O
inhibit	O
a	O
variety	O
of	O
activation-induced	O
events	O
,	O
at	O
least	O
in	O
part	O
by	O
direct	O
interference	O
with	O
AP-1	B-protein
,	O
and	O
is	O
therefore	O
a	O
candidate	O
for	O
a	O
mediator	O
of	O
glucocorticoid-induced	O
immunosuppression	O
.	O

-DOCSTART-	O

Pharmacokinetic	O
differences	O
between	O
a	O
T	O
cell-tolerizing	O
and	O
a	O
T	O
cell-activating	O
peptide	O
.	O

Vaccination	O
with	O
a	O
peptide	O
representing	O
a	O
CTL	O
epitope	O
from	O
the	O
human	O
papillomavirus	O
(	O
HPV	O
)	O
16	O
E7	B-protein
protein	I-protein
induces	O
a	O
specific	O
CTL	O
response	O
that	O
prevents	O
the	O
outgrowth	O
of	O
HPV16	O
E7-expressing	O
tumors	O
.	O

In	O
contrast	O
,	O
vaccination	O
with	O
a	O
peptide	O
encoding	O
an	O
adenovirus	O
type	O
5	O
(	O
Ad5	O
)	O
E1A	O
CTL	O
epitope	O
results	O
in	O
CTL	O
tolerance	O
and	O
enhanced	O
growth	O
of	O
an	O
Ad5	O
E1A-expressing	O
tumor	O
.	O

It	O
is	O
unclear	O
why	O
these	O
peptides	O
induce	O
such	O
opposite	O
effects	O
.	O

To	O
determine	O
whether	O
a	O
difference	O
in	O
pharmacokinetics	O
can	O
explain	O
the	O
functional	O
contrasts	O
,	O
tritiated	O
Ad5	O
E1A	O
and	O
HPV16	O
E7	O
peptides	O
were	O
injected	O
into	O
mice	O
.	O

Results	O
show	O
that	O
the	O
tolerizing	O
peptide	O
spread	O
through	O
the	O
body	O
16	O
times	O
faster	O
than	O
the	O
activating	O
peptide	O
and	O
was	O
cleared	O
at	O
least	O
2	O
times	O
faster	O
.	O

The	O
HPV16	O
E7	O
peptide	O
kinetics	O
correlated	O
with	O
the	O
kinetics	O
of	O
HPV16	B-protein
E7	I-protein
-specific	O
CTL	O
induction	O
.	O

In	O
contrast	O
,	O
Ad5	O
E1A	O
peptide	O
injection	O
resulted	O
in	O
physical	O
deletion	O
of	O
preexisting	O
Ad5	B-protein
E1A	I-protein
-specific	O
CTLs	B-protein
within	O
24	O
h	O
after	O
injection	O
.	O

This	O
tolerization	O
occurred	O
at	O
the	O
time	O
when	O
the	O
peptide	O
reached	O
its	O
maximum	O
peptide	O
concentration	O
in	O
the	O
organs	O
.	O

These	O
data	O
suggest	O
that	O
ubiquitous	O
expression	O
of	O
the	O
tolerizing	O
Ad5	O
E1A	O
peptide	O
within	O
a	O
short	O
period	O
of	O
time	O
causes	O
activation-induced	O
cell	O
death	O
of	O
Ad5	B-protein
E1A	I-protein
-specific	O
CTLs	B-protein
.	O

Therefore	O
,	O
information	O
on	O
the	O
pharmacokinetics	O
of	O
peptides	O
is	O
vital	O
for	O
the	O
safety	O
and	O
efficacy	O
of	O
peptide-based	O
vaccines	O
.	O

-DOCSTART-	O

Smad3	B-protein
and	O
Smad4	B-protein
mediate	O
transforming	B-protein
growth	I-protein
factor-beta1	I-protein
-induced	O
IgA	O
expression	O
in	O
murine	B-cell_type
B	I-cell_type
lymphocytes	I-cell_type
.	O

Transforming	B-protein
growth	I-protein
factor	I-protein
(	I-protein
TGF	I-protein
)	I-protein
-beta1	I-protein
is	O
well	O
established	O
as	O
a	O
critical	O
IgA	B-protein
isotype	I-protein
switching	I-protein
factor	I-protein
and	O
Smad	B-protein
molecules	I-protein
have	O
been	O
reported	O
to	O
act	O
as	O
transducers	B-protein
and	O
transcriptional	B-protein
factors	I-protein
in	O
the	O
expression	O
of	O
TGF-beta1	B-protein
-targeted	O
genes	B-DNA
.	O

We	O
examined	O
the	O
involvement	O
of	O
Smad	B-protein
proteins	I-protein
in	O
TGF-beta1	B-protein
-induced	O
IgA	O
expression	O
.	O

First	O
,	O
we	O
found	O
that	O
TGF-beta1	B-protein
significantly	O
increases	O
endogenous	B-RNA
germ-line	I-RNA
(	I-RNA
GL	I-RNA
)	I-RNA
alpha	I-RNA
transcripts	I-RNA
by	O
LPS-stimulated	O
CH12.LX.4933	B-cell_line
(	I-cell_line
mu	I-cell_line
(	I-cell_line
+	I-cell_line
)	I-cell_line
)	I-cell_line
B	I-cell_line
lymphoma	I-cell_line
cells	I-cell_line
.	O

To	O
investigate	O
its	O
signaling	O
mechanisms	O
,	O
the	O
lymphoma	B-cell_line
cell	I-cell_line
line	I-cell_line
was	O
transfected	O
with	O
pFL3	B-DNA
that	O
contains	O
the	O
TGF-beta-responsive	B-DNA
element	I-DNA
of	O
the	O
GLalpha	B-DNA
promoter	I-DNA
,	O
and	O
stimulated	O
with	O
TGF-beta1	B-protein
.	O

Similar	O
to	O
endogenous	B-RNA
GLalpha	I-RNA
transcripts	I-RNA
,	O
TGF-beta1	B-protein
induces	O
GLalpha	B-DNA
promoter	I-DNA
activity	O
and	O
overexpression	O
of	O
Smad3	B-protein
markedly	O
enhances	O
the	O
promoter	O
activity	O
.	O

This	O
activity	O
is	O
further	O
augmented	O
by	O
cotransfected	B-protein
Smad4	I-protein
.	O

On	O
the	O
other	O
hand	O
,	O
Smad7	B-protein
substantially	O
abrogates	O
the	O
synergistic	O
effect	O
of	O
Smad3/4	B-protein
on	O
GLalpha	B-DNA
promoter	I-DNA
activity	O
.	O

In	O
addition	O
,	O
overexpression	O
of	O
Smad3/4	B-protein
enhances	O
TGF-beta1	B-protein
-induced	O
endogenous	B-RNA
GLalpha	I-RNA
transcripts	I-RNA
in	O
normal	B-cell_type
spleen	I-cell_type
B	I-cell_type
cell	I-cell_type
s	O
.	O

Finally	O
,	O
in	O
the	O
presence	O
of	O
TGF-beta1	B-protein
,	O
overexpression	O
of	O
Smad3/4	B-protein
selectively	O
increases	O
both	O
surface	O
IgA	O
expression	O
and	O
IgA	O
production	O
.	O

The	O
results	O
from	O
the	O
present	O
study	O
indicate	O
that	O
Smad3	B-protein
,	O
Smad4	B-protein
,	O
and	O
Smad7	B-protein
,	O
at	O
least	O
in	O
part	O
,	O
serve	O
as	O
mediators	O
linking	O
TGF-beta1	B-protein
to	O
transcriptional	O
regulation	O
of	O
IgA	B-DNA
switching	I-DNA
related	I-DNA
gene	I-DNA
and	O
regulation	O
of	O
IgA	O
class	O
switching	O
.	O

-DOCSTART-	O

The	O
translesion	B-protein
DNA	I-protein
polymerase	I-protein
zeta	I-protein
plays	O
a	O
major	O
role	O
in	O
Ig	O
and	O
bcl-6	O
somatic	O
hypermutation	O
.	O

Ig	O
somatic	O
mutations	O
would	O
be	O
introduced	O
by	O
a	O
polymerase	B-protein
(	O
pol	B-protein
)	O
while	O
repairing	O
DNA	O
outside	O
main	O
DNA	O
replication	O
.	O

We	O
show	O
that	O
human	B-cell_type
B	I-cell_type
cells	I-cell_type
constitutively	O
express	O
the	O
translesion	B-protein
pol	I-protein
zeta	I-protein
,	O
which	O
effectively	O
extends	O
DNA	O
past	O
mismatched	O
bases	O
(	O
mispair	O
extender	O
)	O
,	O
and	O
pol	B-protein
eta	I-protein
,	O
which	O
bypasses	O
DNA	O
lesions	O
in	O
an	O
error-free	O
fashion	O
.	O

Upon	O
B	B-protein
cell	I-protein
receptor	I-protein
(	O
BCR	B-protein
)	O
engagement	O
and	O
coculture	O
with	O
activated	B-cell_type
CD4+	I-cell_type
T	I-cell_type
cells	I-cell_type
,	O
these	O
lymphocytes	B-cell_type
upregulated	O
pol	B-protein
zeta	I-protein
,	O
downregulated	O
pol	B-protein
eta	I-protein
,	O
and	O
mutated	O
the	O
Ig	B-DNA
and	I-DNA
bcl-6	I-DNA
genes	I-DNA
.	O

Inhibition	O
of	O
the	O
pol	B-protein
zeta	I-protein
REV3	B-protein
catalytic	I-protein
subunit	I-protein
by	O
specific	O
phosphorothioate-modified	O
oligonucleotides	O
impaired	O
Ig	O
and	O
bcl-6	O
hypermutation	O
and	O
UV	O
damage-induced	O
DNA	O
mutagenesis	O
,	O
without	O
affecting	O
cell	O
cycle	O
or	O
viability	O
.	O

Thus	O
,	O
pol	B-protein
zeta	I-protein
plays	O
a	O
critical	O
role	O
in	O
Ig	O
and	O
bcl-6	O
hypermutation	O
,	O
perhaps	O
facilitated	O
by	O
the	O
downregulation	O
of	O
pol	B-protein
eta	I-protein
.	O

-DOCSTART-	O

Molecular	O
mechanism	O
of	O
cell	O
cycle	O
progression	O
induced	O
by	O
the	O
oncogene	B-protein
product	I-protein
Tax	I-protein
of	O
human	O
T-cell	O
leukemia	O
virus	O
type	O
I	O
.	O

The	O
trans-activator	B-protein
protein	I-protein
Tax	I-protein
of	O
human	O
T-cell	O
leukemia	O
virus	O
type	O
I	O
(	O
HTLV-I	O
)	O
plays	O
an	O
important	O
role	O
in	O
the	O
development	O
of	O
adult	O
T-cell	O
leukemia	O
through	O
,	O
at	O
least	O
in	O
part	O
,	O
its	O
ability	O
to	O
stimulate	O
cell	O
growth	O
.	O

We	O
previously	O
reported	O
that	O
Tax	B-protein
induced	O
cell	O
cycle	O
progression	O
from	O
G0/G1	O
phase	O
to	O
S	O
and	O
G2/M	O
phases	O
in	O
human	B-cell_line
T-cell	I-cell_line
line	I-cell_line
Kit	I-cell_line
225	I-cell_line
cells	I-cell_line
.	O

To	O
elucidate	O
molecular	O
mechanism	O
of	O
Tax	B-protein
-induced	O
cell	O
cycle	O
progression	O
,	O
we	O
systematically	O
examined	O
the	O
effects	O
of	O
Tax	B-protein
on	O
biochemical	O
events	O
associated	O
with	O
cell	O
cycle	O
progression	O
.	O

Introduction	O
of	O
Tax	B-protein
into	O
resting	B-cell_line
Kit	I-cell_line
225	I-cell_line
cells	I-cell_line
induced	O
activation	O
of	O
the	O
G1/S	O
transition	O
regulation	O
cascade	O
consisting	O
of	O
activation	O
of	O
cyclin	B-protein
dependent	I-protein
kinase	I-protein
2	I-protein
(	O
CDK2	B-protein
)	O
and	O
CDK4	B-protein
,	O
phosphorylation	O
of	O
the	O
Rb	B-protein
family	I-protein
proteins	I-protein
and	O
an	O
increase	O
in	O
free	O
E2F	B-protein
.	O

The	O
kinase	O
activation	O
was	O
found	O
to	O
result	O
from	O
Tax	B-protein
-induced	O
expression	O
of	O
genes	O
for	O
cell	B-protein
cycle	I-protein
regulatory	I-protein
molecules	I-protein
including	O
cyclin	B-protein
D2	I-protein
,	O
cyclin	B-protein
E	I-protein
,	O
E2F1	B-protein
,	O
CDK2	B-protein
,	O
CDK4	B-protein
and	O
CDK6	B-protein
,	O
and	O
Tax	B-protein
-induced	O
reduction	O
of	O
CDK	B-protein
inhibitors	I-protein
p19	I-protein
(	O
INK4d	B-protein
)	O
and	O
p27	B-protein
(	I-protein
Kip1	I-protein
)	I-protein
.	O

These	O
modulations	O
by	O
Tax	B-protein
always	O
paralleled	O
the	O
ability	O
of	O
Tax	B-protein
to	O
activate	O
the	O
NF-kappaB	O
transcription	O
pathway	O
.	O

These	O
results	O
indicate	O
the	O
important	O
role	O
of	O
Tax	B-protein
-mediated	O
trans-activation	O
of	O
the	O
genes	O
for	O
cell	B-protein
cycle	I-protein
regulatory	I-protein
molecules	I-protein
in	O
Tax	B-protein
-induced	O
cell	O
cycle	O
progression	O
.	O

-DOCSTART-	O

Cot	B-protein
kinase	I-protein
induces	O
cyclooxygenase-2	O
expression	O
in	O
T	B-cell_type
cells	I-cell_type
through	O
activation	O
of	O
the	O
nuclear	B-protein
factor	I-protein
of	I-protein
activated	I-protein
T	I-protein
cells	I-protein
.	O

Cyclooxygenase-2	B-protein
(	O
COX-2	B-protein
)	O
is	O
induced	O
in	O
human	B-cell_type
T	I-cell_type
lymphocytes	I-cell_type
upon	O
T	B-protein
cell	I-protein
receptor	I-protein
triggering	O
.	O

Here	O
we	O
report	O
that	O
Cot	B-protein
kinase	I-protein
,	O
a	O
mitogen-activated	B-protein
protein	I-protein
kinase	I-protein
kinase	I-protein
kinase	I-protein
involved	O
in	O
T	O
cell	O
activation	O
,	O
up-regulates	O
COX-2	B-DNA
gene	I-DNA
expression	O
in	O
Jurkat	B-cell_line
T	I-cell_line
cells	I-cell_line
.	O

Induction	O
of	O
COX-2	O
promoter	O
activity	O
by	O
Cot	B-protein
kinase	I-protein
occurred	O
mainly	O
through	O
activation	O
of	O
the	O
nuclear	B-protein
factor	I-protein
of	I-protein
activated	I-protein
T	I-protein
cells	I-protein
(	O
NFAT	B-protein
)	O
.	O

Mutation	O
of	O
the	O
distal	B-DNA
(	I-DNA
-105/-97	I-DNA
)	I-DNA
and	O
proximal	B-DNA
(	I-DNA
-76/-61	I-DNA
)	I-DNA
NFAT	B-DNA
response	I-DNA
elements	I-DNA
in	O
the	O
COX-2	B-DNA
promoter	I-DNA
abolished	O
the	O
activation	O
induced	O
by	O
Cot	B-protein
kinase	I-protein
.	O

Even	O
more	O
,	O
coexpression	O
of	O
a	O
dominant	O
negative	O
version	O
of	O
NFAT	B-protein
inhibited	O
Cot	B-protein
kinase	I-protein
-mediated	O
COX-2	B-DNA
promoter	I-DNA
activation	O
,	O
whereas	O
cotransfection	O
of	O
a	O
constitutively	O
active	O
version	O
of	O
the	O
calcium-dependent	B-protein
phosphatase	I-protein
calcineurin	I-protein
synergizes	O
with	O
Cot	B-protein
kinase	I-protein
in	O
the	O
up-regulation	O
of	O
COX-2	B-DNA
promoter	I-DNA
-driven	O
transcription	O
.	O

Strikingly	O
,	O
Cot	B-protein
kinase	I-protein
increased	O
transactivation	O
mediated	O
by	O
a	O
GAL4-NFAT	B-protein
fusion	I-protein
protein	I-protein
containing	O
the	O
N-terminal	B-protein
transactivation	I-protein
domain	I-protein
of	O
NFATp	B-protein
.	O

In	O
contrast	O
to	O
phorbol	O
ester	O
plus	O
calcium	O
ionophore	O
A23187	O
,	O
Cot	B-protein
kinase	I-protein
increases	O
both	O
COX-2	B-DNA
promoter	I-DNA
activity	O
and	O
NFAT	B-protein
-mediated	O
transactivation	O
in	O
a	O
cyclosporin	O
A-independent	O
manner	O
.	O

These	O
data	O
indicate	O
that	O
Cot	B-protein
kinase	I-protein
up-regulates	O
COX-2	B-DNA
promoter	I-DNA
-driven	O
transcription	O
through	O
the	O
NFAT	B-DNA
response	I-DNA
elements	I-DNA
,	O
being	O
the	O
Cot	B-protein
kinase	I-protein
-induced	O
NFAT	B-protein
-dependent	O
transactivation	O
presumably	O
implicated	O
in	O
this	O
up-regulation	O
.	O

-DOCSTART-	O

Positive	O
and	O
negative	O
roles	O
of	O
the	O
trans-acting	B-protein
T	I-protein
cell	I-protein
factor-1	I-protein
for	O
the	O
acquisition	O
of	O
distinct	O
Ly-49	B-protein
MHC	I-protein
class	I-protein
I	I-protein
receptors	I-protein
by	O
NK	B-cell_type
cells	I-cell_type
.	O

Members	O
of	O
the	O
Ly-49	B-DNA
gene	I-DNA
family	I-DNA
code	O
for	O
class	B-protein
I	I-protein
MHC-specific	I-protein
receptors	I-protein
that	O
regulate	O
NK	O
cell	O
function	O
.	O

Due	O
to	O
a	O
combinatorial	O
distribution	O
of	O
Ly-49	B-protein
receptors	I-protein
,	O
NK	B-cell_type
cells	I-cell_type
display	O
considerable	O
clonal	O
heterogeneity	O
.	O

The	O
acquisition	O
of	O
one	O
Ly-49	B-protein
receptor	I-protein
,	O
Ly-49A	B-protein
is	O
strictly	O
dependent	O
on	O
the	O
transcriptional	B-protein
trans-acting	I-protein
factor	I-protein
T	B-protein
cell-specific	I-protein
factor-1	I-protein
(	O
TCF-1	B-protein
)	O
.	O

Indeed	O
,	O
TCF-1	B-protein
binds	O
to	O
two	O
sites	O
in	O
the	O
Ly-49a	B-DNA
promoter	I-DNA
and	O
regulates	O
its	O
activity	O
,	O
suggesting	O
that	O
the	O
Ly-49a	B-DNA
gene	I-DNA
is	O
a	O
direct	O
TCF-1	B-protein
target	O
.	O

TCF-1	B-protein
deficiency	O
resulted	O
in	O
the	O
altered	O
usage	O
of	O
additional	O
Ly-49	B-protein
receptors	I-protein
.	O

We	O
show	O
in	O
this	O
study	O
,	O
using	O
TCF-1	B-protein
beta	I-protein
(	I-protein
2	I-protein
)	I-protein
-microglobulin	I-protein
double-deficient	O
mice	O
,	O
that	O
these	O
repertoire	O
alterations	O
are	O
not	O
due	O
to	O
Ly-49/MHC	O
class	O
I	O
interactions	O
.	O

Our	O
findings	O
rather	O
suggest	O
a	O
TCF-1	B-protein
-dependent	O
,	O
cell	O
autonomous	O
effect	O
on	O
the	O
acquisition	O
of	O
multiple	O
Ly-49	B-protein
receptors	I-protein
.	O

Besides	O
reduced	O
receptor	O
usage	O
(	O
Ly-49A	B-protein
and	I-protein
D	I-protein
)	O
,	O
we	O
also	O
observed	O
no	O
effect	O
(	O
Ly-49C	B-protein
)	O
and	O
significantly	O
expanded	O
(	O
Ly-49G	B-protein
and	I-protein
I	I-protein
)	O
receptor	O
usage	O
in	O
the	O
absence	O
of	O
TCF-1	B-protein
.	O

These	O
effects	O
did	O
not	O
in	O
all	O
cases	O
correlate	O
with	O
the	O
presence	O
of	O
TCF	B-DNA
binding	I-DNA
sites	I-DNA
in	O
the	O
respective	O
proximal	B-DNA
promoter	I-DNA
.	O

Therefore	O
,	O
besides	O
TCF-1	B-protein
binding	O
to	O
the	O
proximal	B-DNA
promoter	I-DNA
,	O
Ly-49	O
acquisition	O
may	O
also	O
be	O
regulated	O
by	O
TCF-1	B-protein
binding	O
to	O
more	O
distant	O
cis-acting	B-DNA
elements	I-DNA
and/or	O
by	O
regulating	O
the	O
expression	O
of	O
additional	O
trans-acting	B-DNA
factors	I-DNA
.	O

Consistent	O
with	O
the	O
observed	O
differential	O
,	O
positive	O
or	O
negative	O
role	O
of	O
TCF-1	B-protein
for	O
Ly-49	B-protein
receptor	I-protein
acquisition	O
,	O
reporter	O
gene	O
assays	O
revealed	O
the	O
presence	O
of	O
an	O
inducing	O
as	O
well	O
as	O
a	O
repressing	O
TCF	B-DNA
site	I-DNA
in	O
certain	O
proximal	O
Ly-49	B-DNA
promoters	I-DNA
.	O

These	O
findings	O
reveal	O
an	O
important	O
role	O
of	O
TCF-1	B-protein
for	O
the	O
formation	O
of	O
the	O
NK	B-protein
cell	I-protein
receptor	I-protein
repertoire	O
.	O

-DOCSTART-	O

Ligation	O
of	O
CD11b	B-protein
and	O
CD11c	B-protein
beta	I-protein
(	I-protein
2	I-protein
)	I-protein
integrins	B-protein
by	O
antibodies	B-protein
or	O
soluble	B-protein
CD23	I-protein
induces	O
macrophage	B-protein
inflammatory	I-protein
protein	I-protein
1alpha	I-protein
(	O
MIP-1alpha	B-protein
)	O
and	O
MIP-1beta	B-protein
production	O
in	O
primary	B-cell_type
human	I-cell_type
monocytes	I-cell_type
through	O
a	O
pathway	O
dependent	O
on	O
nuclear	B-protein
factor-kappaB	I-protein
.	O

Chemokines	B-protein
and	O
adhesion	B-protein
molecules	I-protein
such	O
as	O
integrins	B-protein
play	O
a	O
major	O
part	O
in	O
the	O
trafficking	O
,	O
extravasation	O
,	O
and	O
recruitment	O
of	O
leukocytes	B-cell_type
to	O
inflammatory	O
sites	O
.	O

This	O
study	O
investigated	O
the	O
effects	O
of	O
beta	O
(	O
2	O
)	O
integrin	O
engagement	O
on	O
chemokine	O
production	O
by	O
freshly	B-cell_type
isolated	I-cell_type
human	I-cell_type
monocytes	I-cell_type
.	O

We	O
found	O
that	O
ligation	O
of	O
CD11b	B-protein
or	O
CD11c	B-protein
but	O
not	O
CD11a	B-protein
alpha	I-protein
chains	I-protein
of	O
beta	B-protein
(	I-protein
2	I-protein
)	I-protein
integrins	I-protein
by	O
antibodies	B-protein
or	O
soluble	B-protein
CD23	I-protein
(	I-protein
sCD23	I-protein
)	I-protein
fusion	I-protein
proteins	I-protein
rapidly	O
induced	O
transcription	O
and	O
secretion	O
of	O
interleukin	B-protein
8	I-protein
,	O
macrophage	B-protein
inflammatory	I-protein
protein	I-protein
(	I-protein
MIP	I-protein
)	I-protein
1alpha	I-protein
,	O
and	O
MIP-1beta	B-protein
.	O

Because	O
the	O
promoters	B-DNA
of	O
these	O
chemokine	B-DNA
genes	I-DNA
contain	O
kappaB	B-DNA
binding	I-DNA
sites	I-DNA
,	O
we	O
assessed	O
the	O
possible	O
role	O
of	O
nuclear	B-protein
factor-kappaB	I-protein
(	O
NF-kappaB	B-protein
)	O
in	O
controlling	O
induction	O
of	O
the	O
genes	O
through	O
beta	O
(	O
2	O
)	O
integrin	O
engagement	O
.	O

Electrophoretic	O
mobility	O
shift	O
assays	O
showed	O
that	O
sCD23	B-protein
or	O
antibodies	B-protein
to	O
CD11b	B-protein
or	O
to	O
CD11c	B-protein
up-regulated	O
DNA-binding	O
activity	O
of	O
NF-kappaB	B-protein
.	O

Activation	O
of	O
NF-kappaB	B-protein
was	O
accompanied	O
by	O
degradation	O
of	O
its	O
cytosolic	B-protein
inhibitor	I-protein
IkappaB-alpha	I-protein
.	O

Blockade	O
of	O
depletion	O
of	O
IkappaB-alpha	B-protein
by	O
proteasome	O
inhibitors	O
(	O
proteasome	O
inhibitor	O
I	O
or	O
acetyl-leucinyl-leucinyl-norleucinal	O
)	O
led	O
to	O
concomitant	O
inhibition	O
of	O
NF-kappaB	O
DNA-binding	O
activity	O
and	O
expression	O
of	O
MIP-1alpha	B-RNA
and	I-RNA
MIP-1beta	I-RNA
messenger	I-RNA
RNA	I-RNA
induced	O
by	O
beta	O
(	O
2	O
)	O
integrin	O
ligation	O
.	O

These	O
results	O
suggest	O
that	O
triggering	O
of	O
CD11b	B-protein
or	O
CD11c	B-protein
beta	I-protein
(	I-protein
2	I-protein
)	I-protein
integrin	O
on	O
primary	B-cell_type
human	I-cell_type
monocytes	I-cell_type
provides	O
activation	O
signals	O
leading	O
to	O
nuclear	O
translocation	O
of	O
NF-kappaB	B-protein
and	O
subsequent	O
secretion	O
of	O
MIP-1alpha	B-protein
and	O
MIP-1beta	B-protein
that	O
may	O
have	O
an	O
important	O
role	O
in	O
recruitment	O
of	O
other	O
inflammatory	B-cell_type
cells	I-cell_type
during	O
initiation	O
of	O
an	O
inflammatory	O
response	O

-DOCSTART-	O

Synergistic	O
transcriptional	O
activation	O
of	O
human	B-DNA
Acyl-coenzyme	I-DNA
A	I-DNA
:	I-DNA
cholesterol	I-DNA
acyltransterase-1	I-DNA
gene	I-DNA
by	O
interferon-gamma	B-protein
and	O
all-trans-retinoic	O
acid	O
THP-1	B-cell_type
cells	I-cell_type
.	O

Acyl-coenzyme	B-protein
A	I-protein
:	I-protein
cholesterol	I-protein
acyltransferase	I-protein
(	O
ACAT	B-protein
)	O
is	O
an	O
intracellular	B-protein
enzyme	I-protein
involved	O
in	O
cellular	O
cholesterol	O
homeostasis	O
and	O
in	O
atherosclerotic	O
foam	O
cell	O
formation	O
.	O

Human	B-DNA
ACAT-1	I-DNA
gene	I-DNA
contains	O
two	O
promoters	B-DNA
(	O
P1	B-DNA
and	O
P7	B-DNA
)	O
,	O
each	O
located	O
in	O
a	O
different	O
chromosome	B-DNA
(	I-DNA
1	I-DNA
and	I-DNA
7	I-DNA
)	I-DNA
(	O
Li	O
,	O
B.	O
L.	O
,	O
Li	O
,	O
X.	O
L.	O
,	O
Duan	O
,	O
Z.	O
J.	O
,	O
Lee	O
,	O
O.	O
,	O
Lin	O
,	O
S.	O
,	O
Ma	O
,	O
Z.	O
M.	O
,	O
Chang	O
,	O
C.	O
C.	O
,	O
Yang	O
,	O
X.	O
Y.	O
,	O
Park	O
,	O
J.	O
P.	O
,	O
Mohandas	O
,	O
T.	O
K.	O
,	O
Noll	O
,	O
W.	O
,	O
Chan	O
,	O
L.	O
,	O
and	O
Chang	O
,	O
T.	O
Y.	O
(	O
1999	O
)	O
J.	O
Biol	O
Chem.	O
274	O
,	O
11060-11071	O
)	O
.	O

Interferon-gamma	B-protein
(	O
IFN-gamma	B-protein
)	O
,	O
a	O
cytokine	B-protein
that	O
exerts	O
many	O
pro-atherosclerotic	O
effects	O
in	O
vivo	O
,	O
causes	O
up-regulation	O
of	O
ACAT-1	B-RNA
mRNA	I-RNA
in	O
human	B-cell_type
blood	I-cell_type
monocyte-derived	I-cell_type
macrophages	I-cell_type
and	O
macrophage-like	B-cell_type
cells	I-cell_type
but	O
not	O
in	O
other	O
cell	O
types	O
.	O

To	O
examine	O
the	O
molecular	O
nature	O
of	O
this	O
observation	O
,	O
we	O
identified	O
within	O
the	O
ACAT-1	B-DNA
P1	I-DNA
promoter	I-DNA
a	O
159-base	B-DNA
pair	I-DNA
core	I-DNA
region	I-DNA
.	O

This	O
region	O
contains	O
4	O
Sp1	B-DNA
elements	I-DNA
and	O
an	O
IFN-gamma	B-DNA
activated	I-DNA
sequence	I-DNA
(	O
GAS	B-DNA
)	O
that	O
overlaps	O
with	O
the	O
second	O
Sp1	B-DNA
element	I-DNA
.	O

In	O
the	O
monocytic	B-cell_type
cell	I-cell_type
line	I-cell_type
THP-1	I-cell_type
cell	I-cell_type
,	O
the	O
combination	O
of	O
IFN-gamma	B-protein
and	O
all-trans-retinoic	O
acid	O
(	O
a	O
known	O
differentiation	O
agent	O
)	O
enhances	O
the	O
ACAT-1	B-DNA
P1	I-DNA
promoter	I-DNA
but	O
not	O
the	O
P7	B-DNA
promoter	I-DNA
.	O

Additional	O
experiments	O
showed	O
that	O
all-trans-retinoic	O
acid	O
causes	O
large	O
induction	O
of	O
the	O
transcription	B-protein
factor	I-protein
STAT1	I-protein
,	O
while	O
IFN-gamma	B-protein
causes	O
activation	O
of	O
STAT1	B-protein
such	O
that	O
it	O
binds	O
to	O
the	O
GAS/Sp1	B-DNA
site	I-DNA
in	O
the	O
ACAT-1	B-DNA
P1	I-DNA
promoter	I-DNA
.	O

Our	O
work	O
provides	O
a	O
molecular	O
mechanism	O
to	O
account	O
for	O
the	O
effect	O
of	O
IFN-gamma	B-protein
in	O
causing	O
transcriptional	O
activation	O
of	O
ACAT-1	B-protein
in	O
macrophage-like	B-cell_type
cells	I-cell_type
.	O

-DOCSTART-	O

Inhaled	O
nitric	O
oxide	O
down-regulates	O
intrapulmonary	O
nitric	O
oxide	O
production	O
in	O
lipopolysaccharide-induced	O
acute	O
lung	O
injury	O
.	O

OBJECTIVE	O
:	O
To	O
examine	O
whether	O
inhaled	O
nitric	O
oxide	O
(	O
NO	O
)	O
affected	O
the	O
intrapulmonary	O
production	O
of	O
NO	O
,	O
reactive	O
oxygen	O
species	O
,	O
and	O
nuclear	B-protein
factor-kappaB	I-protein
in	O
a	O
lipopolysaccharide	O
(	O
LPS	O
)	O
-induced	O
model	O
of	O
acute	O
lung	O
injury	O
.	O

DESIGN	O
:	O
Prospective	O
,	O
randomized	O
,	O
laboratory	O
study	O
.	O

SETTING	O
:	O
Experimental	O
laboratory	O
at	O
a	O
biomedical	O
institute	O
.	O

SUBJECTS	O
:	O
Twenty	O
male	O
rabbits	O
weighing	O
2.5-3.5	O
kg	O
.	O

INTERVENTIONS	O
:	O
Saline	O
or	O
LPS	O
(	O
5	O
mg/kg	O
of	O
body	O
weight	O
)	O
was	O
administered	O
intravenously	O
with	O
or	O
without	O
NO	O
inhalation	O
(	O
10	O
ppm	O
)	O
in	O
each	O
group	O
of	O
five	O
rabbits	O
.	O

MEASUREMENTS	O
AND	O
MAIN	O
RESULTS	O
:	O
LPS	O
increased	O
the	O
lung	O
leak	O
index	O
,	O
the	O
neutrophils	B-cell_type
and	O
NO	O
levels	O
in	O
bronchoalveolar	O
lavage	O
fluid	O
,	O
and	O
NO	O
levels	O
produced	O
by	O
resting	O
and	O
stimulated	O
alveolar	B-cell_type
macrophages	I-cell_type
.	O

Inhaled	O
NO	O
decreased	O
the	O
lung	O
leak	O
index	O
,	O
the	O
neutrophils	B-cell_type
and	O
NO	O
levels	O
as	O
measured	O
by	O
nitrite	O
levels	O
in	O
the	O
lavage	O
fluid	O
,	O
and	O
NO	O
produced	O
by	O
the	O
resting	O
and	O
stimulated	O
alveolar	B-cell_type
macrophages	I-cell_type
.	O

Inhaled	O
NO	O
also	O
blocked	O
the	O
activities	O
of	O
reactive	O
oxygen	O
species	O
and	O
nuclear	B-protein
factor-kappaB	I-protein
binding	O
to	O
DNA	O
in	O
lavage	B-cell_type
cells	I-cell_type
and	O
in	O
alveolar	B-cell_type
macrophages	I-cell_type
.	O

CONCLUSION	O
:	O
Inhaled	O
NO	O
attenuates	O
LPS-induced	O
acute	O
lung	O
injury	O
,	O
possibly	O
by	O
decreasing	O
NO	O
production	O
in	O
the	O
lungs	O
.	O

The	O
mechanism	O
of	O
reducing	O
NO	O
production	O
resulting	O
from	O
inhaled	O
NO	O
may	O
involve	O
,	O
in	O
part	O
,	O
the	O
activities	O
of	O
reactive	O
oxygen	O
species	O
and/or	O
nuclear	B-protein
factor-kappaB	I-protein
.	O

-DOCSTART-	O

Treatment	O
of	O
allergic	O
airway	O
inflammation	O
and	O
hyperresponsiveness	O
by	O
antisense-induced	O
local	O
blockade	O
of	O
GATA-3	B-DNA
expression	O
.	O

Recent	O
studies	O
in	O
transgenic	O
mice	O
have	O
revealed	O
that	O
expression	O
of	O
a	O
dominant	O
negative	O
form	O
of	O
the	O
transcription	B-protein
factor	I-protein
GATA-3	I-protein
in	O
T	B-cell_type
cells	I-cell_type
can	O
prevent	O
T	B-cell_type
helper	I-cell_type
cell	I-cell_type
type	I-cell_type
2	I-cell_type
(	O
Th2	B-cell_type
)	O
-mediated	O
allergic	O
airway	O
inflammation	O
in	O
mice	O
.	O

However	O
,	O
it	O
remains	O
unclear	O
whether	O
GATA-3	B-protein
plays	O
a	O
role	O
in	O
the	O
effector	O
phase	O
of	O
allergic	O
airway	O
inflammation	O
and	O
whether	O
antagonizing	O
the	O
expression	O
and/or	O
function	O
of	O
GATA-3	B-DNA
can	O
be	O
used	O
for	O
the	O
therapy	O
of	O
allergic	O
airway	O
inflammation	O
and	O
hyperresponsiveness	O
.	O

Here	O
,	O
we	O
analyzed	O
the	O
effects	O
of	O
locally	O
antagonizing	O
GATA-3	B-protein
function	O
in	O
a	O
murine	O
model	O
of	O
asthma	O
.	O

We	O
could	O
suppress	O
GATA-3	O
expression	O
in	O
interleukin	B-protein
(	I-protein
IL	I-protein
)	I-protein
-4	I-protein
-producing	O
T	B-cell_type
cells	I-cell_type
in	O
vitro	O
and	O
in	O
vivo	O
by	O
an	O
antisense	O
phosphorothioate	O
oligonucleotide	O
overlapping	O
the	O
translation	B-DNA
start	I-DNA
site	I-DNA
of	O
GATA-3	B-DNA
,	O
whereas	O
nonsense	O
control	O
oligonucleotides	O
were	O
virtually	O
inactive	O
.	O

In	O
a	O
murine	O
model	O
of	O
asthma	O
associated	O
with	O
allergic	O
pulmonary	O
inflammation	O
and	O
hyperresponsiveness	O
in	O
ovalbumin	O
(	O
OVA	O
)	O
-sensitized	O
mice	O
,	O
local	O
intranasal	O
administration	O
of	O
fluorescein	O
isothiocyanate-labeled	O
GATA-3	O
antisense	O
oligonucleotides	O
led	O
to	O
DNA	O
uptake	O
in	O
lung	B-cell_type
cells	I-cell_type
associated	O
with	O
a	O
reduction	O
of	O
intracellular	O
GATA-3	O
expression	O
.	O

Such	O
intrapulmonary	O
blockade	O
of	O
GATA-3	O
expression	O
caused	O
an	O
abrogation	O
of	O
signs	O
of	O
lung	O
inflammation	O
including	O
infiltration	O
of	O
eosinophils	B-cell_type
and	O
Th2	B-cell_type
cytokine	O
production	O
.	O

Furthermore	O
,	O
treatment	O
with	O
antisense	O
but	O
not	O
nonsense	O
oligonucleotides	O
induced	O
a	O
significant	O
reduction	O
of	O
airway	O
hyperresponsiveness	O
in	O
OVA-sensitized	O
mice	O
to	O
levels	O
comparable	O
to	O
saline-treated	O
control	O
mice	O
,	O
as	O
assessed	O
by	O
both	O
enhanced	O
pause	O
(	O
PenH	O
)	O
responses	O
and	O
pulmonary	O
resistance	O
determined	O
by	O
body	O
plethysmography	O
.	O

These	O
data	O
indicate	O
a	O
critical	O
role	O
for	O
GATA-3	B-DNA
in	O
the	O
effector	O
phase	O
of	O
a	O
murine	O
asthma	O
model	O
and	O
suggest	O
that	O
local	O
delivery	O
of	O
GATA-3	O
antisense	O
oligonucleotides	O
may	O
be	O
a	O
novel	O
approach	O
for	O
the	O
treatment	O
of	O
airway	O
hyperresponsiveness	O
such	O
as	O
in	O
asthma	O
.	O

This	O
approach	O
has	O
the	O
potential	O
advantage	O
of	O
suppressing	O
the	O
expression	O
of	O
various	O
proinflammatory	B-protein
Th2	I-protein
cytokines	I-protein
simultaneously	O
rather	O
than	O
suppressing	O
the	O
activity	O
of	O
a	O
single	O
cytokine	B-protein
.	O

-DOCSTART-	O

T	O
helper-cell	O
phenotype	O
regulates	O
atherosclerosis	O
in	O
mice	O
under	O
conditions	O
of	O
mild	O
hypercholesterolemia	O
.	O

BACKGROUND	O
:	O
T	B-cell_type
cells	I-cell_type
are	O
implicated	O
in	O
atherosclerosis	O
,	O
but	O
little	O
is	O
known	O
about	O
the	O
genetic	O
control	O
or	O
molecular	O
pathways	O
,	O
especially	O
under	O
conditions	O
of	O
mild	O
hypercholesterolemia	O
.	O

METHODS	O
AND	O
RESULTS	O
:	O
BALB/c	O
mice	O
,	O
making	O
a	O
CD4+	O
Th2	O
(	O
IL-4+	O
)	O
cell	O
response	O
,	O
express	O
both	O
MHC	B-protein
class	I-protein
II	I-protein
antigens	I-protein
(	O
IA	B-protein
(	I-protein
d	I-protein
)	I-protein
,	O
IE	B-protein
(	I-protein
d	I-protein
)	I-protein
)	O
and	O
are	O
atherosclerosis-resistant	O
.	O

C57Bl/6	O
mice	O
produce	O
a	O
CD4+	O
Th1	O
(	O
interferon	O
[	O
IFN	O
]	O
gamma+	O
)	O
response	O
,	O
express	O
IA	B-protein
(	I-protein
b	I-protein
)	I-protein
but	O
no	O
IE	B-protein
,	O
and	O
are	O
atherosclerosis-prone	O
.	O

To	O
evaluate	O
T	O
helper-cell	O
phenotype	O
in	O
fatty	O
streak	O
formation	O
,	O
wild-type	O
C57Bl/6	O
mice	O
(	O
IA	O
(	O
b	O
)	O
+IE-	O
)	O
and	O
transgenic	O
mice	O
,	O
either	O
AB	O
(	O
o	O
)	O
,	O
IA	O
(	O
b	O
)	O
-IE-	O
;	O
ABEalpha	O
,	O
IA-IE	O
(	O
k	O
)	O
+	O
;	O
or	O
BL	O
:	O
TG	O
:	O
Ealpha	O
,	O
IA	O
(	O
b	O
)	O
+IE	O
(	O
k	O
)	O
+	O
,	O
were	O
fed	O
a	O
high-cholesterol	O
diet	O
for	O
16	O
weeks	O
and	O
evaluated	O
histomorphometrically	O
for	O
aortic	O
lesions	O
.	O

Lesion	O
size	O
in	O
AB	O
(	O
o	O
)	O
,	O
ABEalpha	O
,	O
and	O
BL	O
:	O
TG	O
:	O
Ealpha	O
strains	O
was	O
decreased	O
by	O
54	O
%	O
,	O
79	O
%	O
,	O
and	O
82	O
%	O
,	O
respectively	O
,	O
compared	O
with	O
wild-type	O
,	O
correlating	O
with	O
decreased	O
Th1	B-cell_type
and	O
increased	O
Th2	B-cell_type
expression	O
and	O
suggesting	O
that	O
T	O
helper-cell	O
phenotype	O
is	O
important	O
in	O
fatty	O
lesion	O
development	O
.	O

Decreasing	O
Th1	B-cell_type
cells	I-cell_type
by	O
antibodies	B-protein
(	O
alpha-CD4	B-protein
)	O
or	O
cytokines	B-protein
(	O
IL-4	B-protein
)	O
also	O
caused	O
>	O
/=80	O
%	O
reductions	O
in	O
lesion	O
size	O
.	O

Immunohistology	O
revealed	O
IFN-gamma	B-protein
,	O
but	O
not	O
IL-4	B-protein
,	O
colocalized	O
with	O
activated	O
macrophages	B-cell_type
.	O

Confirming	O
these	O
findings	O
in	O
a	O
different	O
mouse	O
strain	O
,	O
BALB/c	O
Stat	O
6	O
knockout	O
mice	O
(	O
Th2	O
cell-deficient	O
)	O
developed	O
aortic	O
lesions	O
comparable	O
to	O
C57Bl/6	O
mice	O
on	O
the	O
same	O
diet	O
.	O

CONCLUSIONS	O
:	O
In	O
mildly	O
hypercholesterolemic	O
C57Bl/6	O
mice	O
,	O
presence	O
of	O
IA	B-protein
(	I-protein
b	I-protein
)	I-protein
and	O
absence	O
of	O
IE	B-protein
regulated	O
CD4+	O
T	O
helper-cell	O
phenotype	O
;	O
fatty	O
lesions	O
were	O
proportional	O
to	O
IFNgamma+	B-cell_type
Th1	I-cell_type
cells	I-cell_type
in	O
both	O
C57Bl/6	O
and	O
BALB/c	O
strains	O
.	O

IFN-gamma	B-protein
may	O
participate	O
through	O
macrophage	O
activation	O
,	O
whereas	O
IL-4	B-protein
may	O
act	O
to	O
limit	O
Th1-cell	O
response	O
.	O

-DOCSTART-	O

Requirement	O
for	O
p38	B-protein
and	O
p44/p42	B-protein
mitogen-activated	B-protein
protein	I-protein
kinases	I-protein
in	O
RAGE	B-protein
-mediated	O
nuclear	B-protein
factor-kappaB	I-protein
transcriptional	O
activation	O
and	O
cytokine	O
secretion	O
.	O

Advanced	O
glycation	O
end	O
product	O
(	O
AGE	O
)	O
activation	O
of	O
the	O
signal-transducing	B-protein
receptor	I-protein
for	I-protein
AGE	I-protein
(	O
RAGE	B-protein
)	O
has	O
been	O
linked	O
to	O
a	O
proinflammatory	O
phenotypic	O
change	O
within	O
cells	O
.	O

However	O
,	O
the	O
precise	O
intracellular	O
signaling	O
pathways	O
involved	O
have	O
not	O
been	O
elucidated	O
.	O

We	O
demonstrate	O
here	O
that	O
human	O
serum	O
albumin	O
modified	O
with	O
N	O
(	O
varepsilon	O
)	O
-	O
(	O
carboxymethyl	O
)	O
lysine	O
(	O
CML	O
)	O
,	O
a	O
major	O
AGE	O
adduct	O
that	O
progressively	O
accumulates	O
with	O
aging	O
,	O
diabetes	O
,	O
and	O
renal	O
failure	O
,	O
induced	O
nuclear	B-protein
factor	I-protein
(	I-protein
NF	I-protein
)	I-protein
-kappaB	I-protein
-driven	O
reporter	O
gene	O
expression	O
in	O
human	B-cell_type
monocytic	I-cell_type
THP-1	I-cell_type
cells	I-cell_type
.	O

The	O
NF-kappaB	O
response	O
was	O
blocked	O
with	O
a	O
synthetic	O
peptide	O
corresponding	O
to	O
the	O
putative	B-protein
ligand-binding	I-protein
domain	I-protein
of	O
RAGE	B-protein
,	O
with	O
anti-	O
RAGE	B-protein
antiserum	O
,	O
and	O
by	O
coexpression	O
of	O
truncated	B-protein
receptors	I-protein
lacking	O
the	O
intracellular	B-protein
domain	I-protein
.	O

Signal	O
transduction	O
from	O
RAGE	B-protein
to	O
NF-kappaB	B-protein
involved	O
the	O
generation	O
of	O
reactive	O
oxygen	O
species	O
,	O
since	O
reporter	O
gene	O
expression	O
was	O
blocked	O
with	O
the	O
antioxidant	O
N-acetyl-L-cysteine	O
.	O

CML-modified	B-protein
albumin	I-protein
produced	O
rapid	O
transient	O
activation	O
of	O
tyrosine	O
phosphorylation	O
,	O
extracellular	B-protein
signal-regulated	I-protein
kinase	I-protein
1	I-protein
and	I-protein
2	I-protein
,	O
and	O
p38	B-protein
mitogen-activated	I-protein
protein	I-protein
kinase	I-protein
(	O
MAPK	B-protein
)	O
,	O
but	O
not	O
c-Jun	B-protein
NH	I-protein
(	I-protein
2	I-protein
)	I-protein
-terminal	I-protein
kinase	I-protein
.	O

RAGE	B-protein
-mediated	O
NF-kappaB	O
activation	O
was	O
suppressed	O
by	O
the	O
selective	O
p38	B-protein
MAPK	I-protein
inhibitor	O
SB203580	O
and	O
by	O
coexpression	O
of	O
a	O
kinase-dead	O
p38	O
dominant-negative	O
mutant	O
.	O

Activation	O
of	O
NF-kappaB	B-protein
by	O
CML-modified	B-protein
albumin	I-protein
increased	O
secretion	O
of	O
proinflammatory	B-protein
cytokines	I-protein
(	O
tumor	B-protein
necrosis	I-protein
factor-alpha	I-protein
,	O
interleukin-1beta	B-protein
,	O
and	O
monocyte	B-protein
chemoattractant	I-protein
protein-1	I-protein
)	O
severalfold	O
,	O
and	O
inhibition	O
of	O
p38	B-protein
MAPK	I-protein
blocked	O
these	O
increases	O
.	O

These	O
results	O
indicate	O
that	O
p38	B-protein
MAPK	I-protein
activation	O
mediates	O
RAGE	B-protein
-induced	O
NF-kappaB	B-protein
-dependent	O
secretion	O
of	O
proinflammatory	B-protein
cytokines	I-protein
and	O
suggest	O
that	O
accelerated	O
inflammation	O
may	O
be	O
a	O
consequence	O
of	O
cellular	O
activation	O
induced	O
by	O
this	O
receptor	O
.	O

-DOCSTART-	O

Antigen-receptor	O
cross-linking	O
and	O
lipopolysaccharide	O
trigger	O
distinct	O
phosphoinositide	B-protein
3-kinase	I-protein
-dependent	O
pathways	O
to	O
NF-kappa	O
B	O
activation	O
in	O
primary	B-cell_type
B	I-cell_type
cells	I-cell_type
.	O

The	O
NF-kappaB/Rel	B-protein
transcription	I-protein
factors	I-protein
play	O
an	O
important	O
role	O
in	O
the	O
expression	O
of	O
genes	O
involved	O
in	O
B	O
cell	O
development	O
,	O
differentiation	O
and	O
function	O
.	O

Nuclear	B-protein
NF-kappaB	I-protein
is	O
induced	O
in	O
B	B-cell_type
cells	I-cell_type
by	O
engagement	O
of	O
either	O
the	O
BCR	B-protein
or	O
CD40	B-protein
or	O
by	O
stimulation	O
with	O
lipopolysaccharide	O
(	O
LPS	O
)	O
.	O

Despite	O
the	O
importance	O
of	O
NF-kappaB	B-protein
to	O
B	O
cell	O
function	O
,	O
little	O
is	O
known	O
about	O
the	O
signaling	O
pathways	O
leading	O
to	O
NF-kappaB	O
activation	O
.	O

In	O
this	O
report	O
we	O
address	O
the	O
role	O
of	O
phosphoinositide	B-protein
3'-kinase	I-protein
(	O
PI	B-protein
3-kinase	I-protein
)	O
in	O
BCR	B-protein
-	O
and	O
LPS-induced	O
NF-kappaB	O
activation	O
using	O
populations	O
of	O
primary	B-cell_type
murine	I-cell_type
resting	I-cell_type
B	I-cell_type
cells	I-cell_type
.	O

Using	O
the	O
specific	O
pharmacological	O
inhibitors	O
of	O
PI	B-protein
3-kinase	I-protein
,	O
Wortmannin	O
and	O
LY294002	O
,	O
we	O
demonstrate	O
that	O
PI	B-protein
3-kinase	I-protein
activity	O
is	O
vital	O
for	O
BCR	B-protein
-induced	O
NF-kappaB	B-protein
DNA-binding	O
activity	O
.	O

Furthermore	O
,	O
we	O
show	O
that	O
this	O
is	O
achieved	O
via	O
protein	B-protein
kinase	I-protein
C	I-protein
-dependent	O
degradation	O
of	O
IkappaBalpha	B-protein
.	O

Similar	O
analyses	O
reveal	O
that	O
PI	B-protein
3-kinase	I-protein
is	O
also	O
critical	O
in	O
triggering	O
NF-kappaB	B-protein
DNA-binding	O
activity	O
and	O
IkappaBalpha	B-protein
degradation	O
following	O
LPS	O
stimulation	O
.	O

Interestingly	O
,	O
a	O
PKC	O
inhibitor	O
which	O
blocked	O
the	O
BCR	B-protein
-induced	O
IkappaBalpha	B-protein
degradation	O
had	O
no	O
effect	O
on	O
the	O
degradation	O
of	O
IkappaBalpha	B-protein
after	O
LPS	O
stimulation	O
.	O

Taken	O
together	O
,	O
our	O
results	O
indicate	O
the	O
involvement	O
of	O
PI	B-protein
3-kinase	I-protein
in	O
at	O
least	O
two	O
distinct	O
signaling	O
pathways	O
leading	O
to	O
activation	O
of	O
NF-kappaB	B-protein
in	O
B	B-cell_type
cells	I-cell_type
.	O

-DOCSTART-	O

Tetramer-guided	O
epitope	O
mapping	O
:	O
rapid	O
identification	O
and	O
characterization	O
of	O
immunodominant	B-cell_type
CD4+	I-cell_type
T	I-cell_type
cell	I-cell_type
epitopes	I-cell_type
from	O
complex	O
antigens	B-protein
.	O

T	O
cell	O
responses	O
to	O
Ags	B-protein
involve	O
recognition	O
of	O
selected	O
peptide	B-protein
epitopes	I-protein
contained	O
within	O
the	O
antigenic	B-protein
protein	I-protein
.	O

In	O
this	O
report	O
,	O
we	O
describe	O
a	O
new	O
approach	O
for	O
direct	O
identification	O
of	O
CD4+	B-cell_type
T	I-cell_type
cell	I-cell_type
epitopes	I-cell_type
of	O
complex	O
Ags	B-protein
that	O
uses	O
human	B-protein
class	I-protein
II	I-protein
tetramers	I-protein
to	O
identify	O
reactive	O
cells	O
.	O

With	O
a	O
panel	O
of	O
60	O
overlapping	O
peptides	O
covering	O
the	O
entire	O
sequence	O
of	O
the	O
VP16	B-protein
protein	I-protein
,	O
a	O
major	O
Ag	B-protein
for	O
HSV-2	O
,	O
we	O
generated	O
a	O
panel	O
of	O
class	B-protein
II	I-protein
MHC	I-protein
tetramers	I-protein
loaded	O
with	O
peptide	O
pools	O
that	O
were	O
used	O
to	O
stain	O
peripheral	B-cell_type
lymphocytes	I-cell_type
of	O
an	O
HSV-2	O
infected	O
individual	O
.	O

With	O
this	O
approach	O
,	O
we	O
identified	O
four	O
new	O
DRA1*0101/DRB1*0401-	O
and	O
two	O
DRA1*0101/DRB1*0404-restricted	O
,	O
VP16-specific	B-protein
epitopes	I-protein
.	O

By	O
using	O
tetramers	B-protein
to	O
sort	O
individual	O
cells	O
,	O
we	O
easily	O
obtained	O
a	O
large	O
number	O
of	O
clones	O
specific	O
to	O
these	O
epitopes	B-protein
.	O

Although	O
DRA1*0101/DRB1*0401	O
and	O
DRA1*0101/DRB1*0404	O
are	O
structurally	O
very	O
similar	O
,	O
nonoverlapping	O
VP16	B-protein
epitopes	I-protein
were	O
identified	O
,	O
illustrating	O
high	O
selectivity	O
of	O
individual	O
allele	O
polymorphisms	O
within	O
common	O
MHC	O
variants	O
.	O

This	O
rapid	O
approach	O
to	O
detecting	O
CD4+	B-cell_type
T	I-cell_type
cell	I-cell_type
epitopes	I-cell_type
from	O
complex	O
Ags	B-protein
can	O
be	O
applied	O
to	O
any	O
known	O
Ag	B-protein
that	O
gives	O
a	O
T	O
cell	O
response	O
.	O

-DOCSTART-	O

Localized	O
pancreatic	O
NF-kappaB	O
activation	O
and	O
inflammatory	O
response	O
in	O
taurocholate-induced	O
pancreatitis	O
.	O

Transcription	B-protein
factor	I-protein
nuclear	I-protein
factor-kappaB	I-protein
(	O
NF-kappaB	B-protein
)	O
is	O
activated	O
in	O
cerulein	O
pancreatitis	O
and	O
mediates	O
cytokine	O
expression	O
.	O

The	O
role	O
of	O
transcription	O
factor	O
activation	O
in	O
other	O
models	O
of	O
pancreatitis	O
has	O
not	O
been	O
established	O
.	O

Here	O
we	O
report	O
upregulation	O
of	O
NF-kappaB	B-protein
and	O
inflammatory	B-protein
molecules	I-protein
,	O
and	O
their	O
correlation	O
with	O
local	O
pancreatic	O
injury	O
,	O
in	O
a	O
model	O
of	O
severe	O
pancreatitis	O
.	O

Rats	O
received	O
intraductal	O
infusion	O
of	O
taurocholate	O
or	O
saline	O
,	O
and	O
the	O
pancreatic	O
head	O
and	O
tail	O
were	O
analyzed	O
separately	O
.	O

NF-kappaB	B-protein
and	O
activator	B-protein
protein-1	I-protein
(	O
AP-1	B-protein
)	O
activation	O
were	O
assessed	O
by	O
gel	O
shift	O
assay	O
,	O
and	O
mRNA	O
expression	O
of	O
interleukin-6	B-RNA
,	O
tumor	B-protein
necrosis	I-protein
factor-alpha	I-protein
,	O
KC	O
,	O
monocyte	B-protein
chemoattractant	I-protein
protein-1	I-protein
,	O
and	O
inducible	B-protein
nitric	I-protein
oxide	I-protein
synthase	I-protein
was	O
assessed	O
by	O
semiquantitative	O
RT-PCR	O
.	O

Morphological	O
damage	O
and	O
trypsin	O
activation	O
were	O
much	O
greater	O
in	O
the	O
pancreatic	O
head	O
than	O
tail	O
,	O
in	O
parallel	O
with	O
a	O
stronger	O
activation	O
of	O
NF-kappaB	B-RNA
and	I-RNA
cytokine	I-RNA
mRNA	I-RNA
.	O

Saline	O
infusion	O
mildly	O
affected	O
these	O
parameters	O
.	O

AP-1	B-protein
was	O
strongly	O
activated	O
in	O
both	O
pancreatic	O
segments	O
after	O
either	O
taurocholate	O
or	O
saline	O
infusion	O
.	O

NF-kappaB	B-protein
inhibition	O
with	O
N-acetylcysteine	O
ameliorated	O
the	O
local	O
inflammatory	O
response	O
.	O

Correlation	O
between	O
localized	O
NF-kappaB	B-protein
activation	O
,	O
cytokine	O
upregulation	O
,	O
and	O
tissue	O
damage	O
suggests	O
a	O
key	O
role	O
for	O
NF-kappaB	B-protein
in	O
the	O
development	O
of	O
the	O
inflammatory	O
response	O
of	O
acute	O
pancreatitis	O
.	O

-DOCSTART-	O

CD45	B-protein
tyrosine	I-protein
phosphatase	I-protein
controls	O
common	O
gamma-chain	B-protein
cytokine	I-protein
-mediated	O
STAT	B-protein
and	O
extracellular	O
signal-related	O
kinase	O
phosphorylation	O
in	O
activated	B-cell_type
human	I-cell_type
lymphoblasts	I-cell_type
:	O
inhibition	O
of	O
proliferation	O
without	O
induction	O
of	O
apoptosis	O
.	O

The	O
objective	O
of	O
this	O
study	O
was	O
to	O
test	O
whether	O
CD45	O
signals	O
can	O
influence	O
signaling	O
processes	O
in	O
activated	B-cell_type
human	I-cell_type
lymphoblasts	I-cell_type
.	O

To	O
this	O
end	O
,	O
we	O
generated	O
lymphoblasts	B-cell_type
which	O
proliferate	O
in	O
response	O
to	O
common	O
gamma-chain	B-protein
cytokines	I-protein
,	O
but	O
readily	O
undergo	O
apoptosis	O
after	O
cytokine	O
withdrawal	O
.	O

In	O
experiments	O
with	O
the	O
CD45R0	B-protein
mAb	I-protein
UCHL-1	I-protein
,	O
but	O
not	O
control	B-protein
CD45	I-protein
mAbs	I-protein
,	O
we	O
found	O
significant	O
inhibition	O
of	O
proliferation	O
.	O

Interestingly	O
,	O
the	O
pan-CD45	B-protein
mAb	I-protein
GAP8.3	I-protein
,	O
which	O
is	O
most	O
effective	O
in	O
inhibition	O
of	O
OKT-3-mediated	O
proliferation	O
in	O
quiescent	B-cell_type
lymphocytes	I-cell_type
,	O
was	O
ineffective	O
in	O
lymphoblasts	B-cell_type
.	O

Addition	O
of	O
CD3	B-protein
mAb	I-protein
OKT-3	I-protein
had	O
no	O
influence	O
on	O
IL-2-mediated	O
proliferation	O
(	O
with	O
or	O
without	O
UCHL-1	B-protein
)	O
.	O

In	O
contrast	O
,	O
after	O
addition	O
of	O
OKT-3	B-protein
to	O
IL-4	B-protein
-	O
and	O
IL-7	B-protein
-stimulated	O
proliferation	O
assays	O
,	O
UCHL-1	B-protein
signals	O
could	O
not	O
significantly	O
alter	O
cellular	O
proliferation	O
.	O

We	O
did	O
not	O
find	O
induction	O
of	O
apoptosis	O
following	O
CD45R0	O
signaling	O
.	O

In	O
Western	O
blots	O
using	O
mAbs	B-protein
detecting	O
phosphorylated	B-protein
STAT-3	I-protein
,	I-protein
STAT-5	I-protein
,	I-protein
STAT-6	I-protein
,	O
or	O
extracellular	B-protein
signal-related	I-protein
kinase	I-protein
1/2	I-protein
,	O
we	O
found	O
that	O
CD45R0	O
signaling	O
could	O
effectively	O
diminish	O
phosphorylation	O
of	O
these	O
intracellular	O
signaling	O
components	O
.	O

Using	O
RT-PCR	O
,	O
we	O
found	O
that	O
CD45R0	O
signaling	O
inhibited	O
IL-2	O
mRNA	O
production	O
without	O
major	O
influence	O
on	O
IL-13	B-protein
,	O
IL-5	B-protein
,	O
or	O
IFN-gamma	B-RNA
mRNA	I-RNA
levels	O
.	O

Costimulation	O
with	O
OKT-3	B-protein
and	O
IL-2	B-protein
optimally	O
induced	O
secretion	O
of	O
IFN-gamma	B-protein
,	O
TNF-alpha	B-protein
,	O
and	O
IL-5	B-protein
,	O
which	O
was	O
not	O
decreased	O
by	O
CD45	O
signals	O
.	O

In	O
conclusion	O
,	O
we	O
illustrate	O
that	O
CD45R0	O
signals	O
control	O
early	O
cytokine	O
receptor-associated	O
signaling	O
processes	O
and	O
mRNA	O
and	O
DNA	O
synthesis	O
in	O
activated	B-cell_type
human	I-cell_type
lymphoblasts	I-cell_type
.	O

Furthermore	O
,	O
we	O
show	O
the	O
existence	O
of	O
CD45	B-protein
epitopes	I-protein
(	O
GAP8.3	B-protein
)	O
,	O
which	O
are	O
active	O
and	O
critical	O
for	O
signaling	O
in	O
quiescent	B-cell_type
lymphocytes	I-cell_type
,	O
but	O
are	O
nonfunctional	O
in	O
activated	B-cell_type
human	I-cell_type
lymphoblasts	I-cell_type
.	O

-DOCSTART-	O

Pax5	B-DNA
determines	O
the	O
identity	O
of	O
B	B-cell_type
cells	I-cell_type
from	O
the	O
beginning	O
to	O
the	O
end	O
of	O
B-lymphopoiesis	O
.	O

Despite	O
being	O
one	O
of	O
the	O
most	O
intensively	O
studied	O
cell	O
types	O
,	O
the	O
molecular	O
basis	O
of	O
B	O
cell	O
specification	O
is	O
largely	O
unknown	O
.	O

The	O
Pax5	B-DNA
gene	I-DNA
encoding	O
the	O
transcription	B-protein
factor	I-protein
BSAP	I-protein
is	O
required	O
for	O
progression	O
of	O
B-lymphopoiesis	O
beyond	O
the	O
pro-B	O
cell	O
stage	O
.	O

Pax5-deficient	B-cell_type
pro-B	I-cell_type
cells	I-cell_type
are	O
,	O
however	O
,	O
not	O
yet	O
committed	O
to	O
the	O
B-lymphoid	B-cell_type
lineage	I-cell_type
,	O
but	O
instead	O
have	O
a	O
broad	O
lymphomyeloid	O
developmental	O
potential	O
.	O

Pax5	B-DNA
appears	O
to	O
mediate	O
B-lineage	O
commitment	O
by	O
repressing	O
the	O
transcription	O
of	O
non-B-lymphoid	B-DNA
genes	I-DNA
and	O
by	O
simultaneously	O
activating	O
the	O
expression	O
of	O
B-lineage-specific	B-DNA
genes	I-DNA
.	O

Pax5	B-DNA
thus	O
functions	O
both	O
as	O
a	O
transcriptional	O
repressor	O
and	O
activator	O
,	O
depending	O
on	O
its	O
interactions	O
with	O
corepressors	B-protein
of	O
the	O
Groucho	B-protein
protein	I-protein
family	I-protein
or	O
with	O
positive	O
regulators	O
such	O
as	O
the	O
TATA-binding	B-protein
protein	I-protein
.	O

Once	O
committed	O
to	O
the	O
B-lineage	B-cell_type
,	O
B	B-cell_type
cells	I-cell_type
require	O
Pax5	B-DNA
function	O
to	O
maintain	O
their	O
B-lymphoid	O
identity	O
throughout	O
B	O
cell	O
development	O

-DOCSTART-	O

Partners	O
in	O
transcription	O
:	O
NFAT	B-protein
and	O
AP-1	B-protein
.	O

Combinatorial	O
regulation	O
is	O
a	O
powerful	O
mechanism	O
that	O
enables	O
tight	O
control	O
of	O
gene	O
expression	O
,	O
via	O
integration	O
of	O
multiple	O
signaling	O
pathways	O
that	O
induce	O
different	O
transcription	B-protein
factors	I-protein
required	O
for	O
enhanceosome	O
assembly	O
.	O

The	O
four	O
calcium-regulated	O
transcription	B-protein
factors	I-protein
of	O
the	O
NFAT	B-protein
family	I-protein
act	O
synergistically	O
with	O
AP-1	B-protein
(	I-protein
Fos/Jun	I-protein
)	I-protein
proteins	I-protein
on	O
composite	O
DNA	O
elements	O
which	O
contain	O
adjacent	O
NFAT	B-DNA
and	I-DNA
AP-1	I-DNA
binding	I-DNA
sites	I-DNA
,	O
where	O
they	O
form	O
highly	O
stable	O
ternary	O
complexes	O
to	O
regulate	O
the	O
expression	O
of	O
diverse	O
inducible	O
genes	O
.	O

Concomitant	O
induction	O
of	O
NFAT	B-protein
and	O
AP-1	B-protein
requires	O
concerted	O
activation	O
of	O
two	O
different	O
signaling	O
pathways	O
:	O
calcium/calcineurin	O
,	O
which	O
promotes	O
NFAT	O
dephosphorylation	O
,	O
nuclear	O
translocation	O
and	O
activation	O
;	O
and	O
protein	B-protein
kinase	I-protein
C	I-protein
(	I-protein
PKC	I-protein
)	I-protein
/Ras	I-protein
,	O
which	O
promotes	O
the	O
synthesis	O
,	O
phosphorylation	O
and	O
activation	O
of	O
members	O
of	O
the	O
Fos	B-protein
and	I-protein
Jun	I-protein
families	I-protein
of	O
transcription	B-protein
factors	I-protein
.	O

A	O
fifth	O
member	O
of	O
the	O
NFAT	B-protein
family	I-protein
,	O
NFAT5	B-protein
,	O
controls	O
the	O
cellular	O
response	O
to	O
osmotic	O
stress	O
,	O
by	O
a	O
mechanism	O
that	O
requires	O
dimer	O
formation	O
and	O
is	O
independent	O
of	O
calcineurin	B-protein
or	O
of	O
interaction	O
with	O
AP-1	B-protein
.	O

Pharmacological	O
interference	O
with	O
theNFAT	O
:	O
AP-1	O
interaction	O
may	O
be	O
useful	O
in	O
selective	O
manipulation	O
of	O
the	O
immune	O
response	O
.	O

Balanced	O
activation	O
of	O
NFAT	B-protein
and	O
AP-1	B-protein
is	O
known	O
to	O
be	O
required	O
for	O
productive	O
immune	O
responses	O
,	O
but	O
the	O
role	O
of	O
NFAT	O
:	O
AP-1	O
interactions	O
in	O
other	O
cell	O
types	O
and	O
biological	O
processes	O
remains	O
to	O
be	O
understood	O
.	O

-DOCSTART-	O

Cytokine	O
production	O
by	O
Vgamma	B-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
-T-cell	I-cell_type
subsets	I-cell_type
is	O
an	O
important	O
factor	O
determining	O
CD4	O
(	O
+	O
)	O
-Th-cell	O
phenotype	O
and	O
susceptibility	O
of	O
BALB/c	O
mice	O
to	O
coxsackievirus	O
B3-induced	O
myocarditis	O
.	O

Two	O
coxsackievirus	O
B3	O
(	O
CVB3	O
)	O
variants	O
(	O
H3	O
and	O
H310A1	O
)	O
differ	O
by	O
a	O
single	O
amino	O
acid	O
mutation	O
in	O
the	O
VP2	B-protein
capsid	I-protein
protein	I-protein
.	O

H3	O
induces	O
severe	O
myocarditis	O
in	O
BALB/c	O
mice	O
,	O
but	O
H310A1	O
is	O
amyocarditic	O
.	O

Infection	O
with	O
H3	O
,	O
but	O
not	O
H310A1	O
,	O
preferentially	O
activates	O
Vgamma4	B-cell_type
Vdelta4	I-cell_type
cells	I-cell_type
,	O
which	O
are	O
strongly	O
positive	O
for	O
gamma	B-protein
interferon	I-protein
(	O
IFN-gamma	B-protein
)	O
,	O
whereas	O
Vgamma1	B-cell_type
Vdelta4	I-cell_type
cells	I-cell_type
are	O
increased	O
in	O
both	O
H3	O
and	O
H310A1	O
virus-infected	O
animals	O
.	O

Depletion	O
of	O
Vgamma1	B-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
cells	I-cell_type
using	O
monoclonal	B-protein
anti-Vgamma1	I-protein
antibody	I-protein
enhanced	O
myocarditis	O
and	O
CD4	O
(	O
+	O
)	O
-	O
,	O
IFN-gamma	B-protein
(	O
+	O
)	O
-cell	O
responses	O
in	O
both	O
H3-	O
and	O
H310A1-infected	O
mice	O
yet	O
decreased	O
the	O
CD4	O
(	O
+	O
)	O
-	O
,	O
IL-4	O
(	O
+	O
)	O
-cell	O
response	O
.	O

Depleting	O
Vgamma4	B-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
cells	I-cell_type
suppressed	O
myocarditis	O
and	O
reduced	O
CD4	B-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
IFN-gamma	I-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
cells	I-cell_type
but	O
increased	O
CD4	B-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
IL-4	I-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
T	I-cell_type
cells	I-cell_type
.	O

The	O
role	O
of	O
cytokine	O
production	O
by	O
Vgamma1	B-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
and	I-cell_type
Vgamma4	I-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
T	I-cell_type
cells	I-cell_type
was	O
investigated	O
by	O
adoptively	O
transferring	O
these	O
cells	O
isolated	O
from	O
H3-infected	O
BALB/c	O
Stat4	O
knockout	O
(	O
Stat4ko	O
)	O
(	O
defective	O
in	O
IFN-gamma	B-protein
expression	O
)	O
or	O
BALB/c	O
Stat6ko	O
(	O
defective	O
in	O
IL-4	O
expression	O
)	O
mice	O
into	O
H3	O
virus-infected	O
wild-type	O
BALB/c	O
recipients	O
.	O

Vgamma4	B-cell_type
and	I-cell_type
Vgamma1	I-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
T	I-cell_type
cells	I-cell_type
from	O
Stat4ko	O
mice	O
expressed	O
IL-4	B-protein
but	O
no	O
or	O
minimal	O
IFN-gamma	B-protein
,	O
whereas	O
these	O
cell	O
populations	O
derived	O
from	O
Stat6ko	O
mice	O
expressed	O
IFN-gamma	B-protein
but	O
no	O
IL-4	B-protein
.	O

Stat4ko	B-cell_type
Vgamma1	I-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
cells	I-cell_type
(	I-cell_type
IL-4	I-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
)	O
suppress	O
myocarditis	O
.	O

Stat6ko	B-cell_type
Vgamma1	I-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
cells	I-cell_type
(	I-cell_type
IFN-gamma	I-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
)	I-cell_type
were	O
not	O
inhibitory	O
.	O

Stat6ko	B-cell_type
Vgamma4	I-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
cells	I-cell_type
(	I-cell_type
IFN-gamma	I-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
)	I-cell_type
significantly	O
enhanced	O
myocarditis	O
.	O

Stat4ko	B-cell_type
Vgamma4	I-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
cells	I-cell_type
(	I-cell_type
IL-4	I-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
)	I-cell_type
neither	O
inhibited	O
nor	O
enhanced	O
disease	O
.	O

These	O
results	O
show	O
that	O
distinct	O
gammadelta-T-cell	B-cell_type
subsets	I-cell_type
control	O
myocarditis	O
susceptibility	O
and	O
bias	O
the	O
CD4	B-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
-Th-cell	I-cell_type
response	O
.	O

The	O
cytokines	B-protein
produced	O
by	O
the	O
Vgamma	B-cell_type
subpopulation	I-cell_type
have	O
a	O
significant	O
influence	O
on	O
the	O
CD4	B-protein
(	O
+	O
)	O
-Th-cell	O
phenotype	O
.	O

-DOCSTART-	O

Plasmin	B-protein
-induced	O
expression	O
of	O
cytokines	B-protein
and	O
tissue	B-protein
factor	I-protein
in	O
human	B-cell_type
monocytes	I-cell_type
involves	O
AP-1	B-protein
and	O
IKKbeta	B-protein
-mediated	O
NF-kappaB	O
activation	O
.	O

It	O
was	O
previously	O
shown	O
that	O
plasmin	B-protein
activates	O
human	B-cell_type
peripheral	I-cell_type
monocytes	I-cell_type
in	O
terms	O
of	O
lipid	O
mediator	O
release	O
and	O
chemotactic	O
migration	O
.	O

Here	O
it	O
is	O
demonstrated	O
that	O
plasmin	B-protein
induces	O
proinflammatory	B-protein
cytokine	I-protein
release	O
and	O
tissue	B-protein
factor	I-protein
(	O
TF	B-protein
)	O
expression	O
by	O
monocytes	B-cell_type
.	O

Plasmin	B-protein
0.043	O
to	O
1.43	O
CTA	O
U/mL	O
,	O
but	O
not	O
active	B-protein
site-blocked	I-protein
plasmin	I-protein
,	O
triggered	O
concentration-dependent	O
expression	O
of	O
mRNA	O
for	O
interleukin-1alpha	B-protein
(	O
IL-1alpha	B-protein
)	O
,	O
IL-1beta	B-protein
,	O
tumor	B-protein
necrosis	I-protein
factor-alpha	I-protein
(	O
TNF-alpha	B-protein
)	O
,	O
and	O
TF	B-protein
with	O
maximum	O
responses	O
after	O
4	O
hours	O
.	O

Plasmin	B-protein
-mediated	O
mRNA	O
expression	O
was	O
inhibited	O
in	O
a	O
concentration-dependent	O
manner	O
by	O
the	O
lysine	O
analogue	O
trans-4-	O
(	O
aminomethyl	O
)	O
cyclohexane-1-carboxylic	O
acid	O
(	O
t-AMCA	O
)	O
.	O

Increases	O
in	O
mRNA	O
levels	O
were	O
followed	O
by	O
concentration-	O
and	O
time-dependent	O
release	O
of	O
IL-1alpha	B-protein
,	O
IL-1beta	B-protein
and	O
TNF-alpha	B-protein
and	O
by	O
TF	B-protein
expression	O
on	O
monocyte	O
surfaces	O
.	O

Neither	O
cytokines	B-protein
nor	O
TF	B-protein
could	O
be	O
detected	O
when	O
monocytes	B-cell_type
were	O
preincubated	O
with	O
actinomycin	O
D	O
or	O
cycloheximide	O
.	O

Electrophoretic	O
mobility	O
shift	O
assays	O
indicated	O
plasmin	B-protein
-induced	O
activation	O
of	O
NF-kappaB	B-protein
;	O
DNA-binding	B-protein
complexes	I-protein
were	O
composed	O
of	O
p50	B-protein
,	O
p65	B-protein
,	O
and	O
c-Rel	B-protein
,	O
as	O
shown	O
by	O
supershift	O
experiments	O
.	O

Nuclear	O
translocation	O
of	O
NF-kappaB/Rel	B-protein
proteins	I-protein
coincided	O
with	O
IkappaBalpha	O
degradation	O
.	O

At	O
variance	O
with	O
endotoxic	O
lipopolysaccharide	O
,	O
plasmin	B-protein
elicited	O
the	O
rapid	O
degradation	O
of	O
another	O
cytoplasmic	B-protein
NF-kappaB	I-protein
inhibitor	I-protein
,	I-protein
p105	I-protein
.	O

Proteolysis	O
of	O
NF-kappaB	O
inhibitors	O
was	O
apparently	O
due	O
to	O
transient	O
activation	O
of	O
IkappaB	B-protein
kinase	I-protein
(	I-protein
IKK	I-protein
)	I-protein
beta	I-protein
that	O
reached	O
maximum	O
activity	O
at	O
1	O
hour	O
after	O
plasmin	B-protein
stimulation	O
.	O

In	O
addition	O
,	O
AP-1	O
binding	O
was	O
increased	O
in	O
plasmin	B-protein
-treated	O
monocytes	B-cell_type
,	O
with	O
most	O
complexes	O
composed	O
of	O
JunD	B-protein
,	O
c-Fos	B-protein
,	O
and	O
FosB	B-protein
.	O

These	O
findings	O
further	O
substantiate	O
the	O
role	O
of	O
plasmin	B-protein
as	O
a	O
proinflammatory	O
activator	O
of	O
human	B-cell_type
monocytes	I-cell_type
and	O
reveal	O
an	O
important	O
new	O
link	O
between	O
the	O
plasminogen-plasmin	O
system	O
and	O
inflammation	O
.	O

(	O
Blood.	O
2001	O
;	O
97	O
:	O
3941-3950	O
)	O

-DOCSTART-	O

STAT3	B-protein
is	O
constitutively	O
active	O
in	O
some	O
patients	O
with	O
Polycythemia	O
rubra	O
vera	O
.	O

OBJECTIVE	O
:	O
Polycythemia	O
vera	O
is	O
a	O
clonal	O
stem	O
cell	O
disorder	O
characterized	O
by	O
hyperproliferation	O
of	O
the	O
erythroid	B-cell_type
,	I-cell_type
myeloid	I-cell_type
,	I-cell_type
and	I-cell_type
megakaryocytic	I-cell_type
lineages	I-cell_type
.	O

While	O
it	O
has	O
been	O
shown	O
that	O
progenitor	B-cell_type
cells	I-cell_type
of	O
P.	O
vera	O
patients	O
are	O
hypersensitive	O
to	O
several	O
growth	O
factors	O
including	O
erythropoietin	B-protein
,	O
insulin-like	B-protein
growth	I-protein
factor-1	I-protein
,	O
thrombopoietin	B-protein
,	O
interleukin-3	B-protein
,	O
and	O
granulocyte/monocyte	B-protein
colony-stimulating	I-protein
factor	I-protein
,	O
the	O
molecular	O
pathogenesis	O
of	O
this	O
disease	O
remains	O
unknown	O
.	O

Growth	O
factor	O
hypersensitivity	O
could	O
be	O
mediated	O
by	O
changes	O
in	O
signal	O
transduction	O
pathways	O
.	O

We	O
therefore	O
investigated	O
a	O
common	O
downstream	O
effector	O
of	O
cytokines	B-protein
,	O
the	O
signal	B-protein
transducers	I-protein
and	I-protein
activators	I-protein
of	I-protein
transcription	I-protein
(	O
STATs	B-protein
)	O
.	O

A	O
constitutive	O
activation	O
of	O
STAT	B-protein
factors	I-protein
could	O
explain	O
the	O
increased	O
proliferation	O
of	O
P.	B-cell_type
vera	I-cell_type
cells	I-cell_type
even	O
in	O
the	O
absence	O
of	O
growth	O
factor	O
stimulation	O
.	O

METHODS	O
:	O
Peripheral	B-cell_type
granulocytes	I-cell_type
from	O
patients	O
with	O
P.	O
vera	O
and	O
from	O
healthy	O
volunteers	O
were	O
assayed	O
for	O
STAT1	B-protein
,	I-protein
3	I-protein
,	I-protein
and	I-protein
5	I-protein
DNA	O
binding	O
by	O
electrophoretic	O
mobility	O
shift	O
assay	O
.	O

RESULTS	O
:	O
Four	O
of	O
14	O
P.	O
vera	O
patients	O
analyzed	O
showed	O
constitutive	O
STAT3	O
DNA	O
binding	O
in	O
unstimulated	B-cell_type
peripheral	I-cell_type
granulocytes	I-cell_type
,	O
while	O
none	O
of	O
the	O
17	O
healthy	O
volunteers	O
tested	O
did	O
.	O

None	O
of	O
the	O
subjects	O
showed	O
constitutive	O
STAT1	B-protein
or	O
STAT5	B-protein
activity	O
.	O

Western	O
blotting	O
demonstrated	O
that	O
,	O
in	O
the	O
three	O
patients	O
,	O
STAT3	B-protein
is	O
constitutively	O
phosphorylated	O
on	O
Tyr	O
705	O
,	O
whereas	O
it	O
is	O
unphosphorylated	O
in	O
the	O
other	O
patients	O
and	O
in	O
controls	O
.	O

Interestingly	O
,	O
constitutive	O
STAT3	B-protein
activity	O
did	O
not	O
correlate	O
with	O
the	O
duration	O
of	O
disease	O
or	O
the	O
treatment	O
regimen	O
.	O

It	O
was	O
observed	O
in	O
a	O
recently	O
diagnosed	O
patient	O
and	O
in	O
two	O
patients	O
treated	O
only	O
with	O
phlebotomy	O
.	O

CONCLUSION	O
:	O
Our	O
data	O
suggest	O
that	O
constitutive	O
phosphorylation	O
and	O
activation	O
of	O
STAT3	B-protein
is	O
not	O
a	O
secondary	O
event	O
induced	O
by	O
mutagenizing	O
agents	O
or	O
by	O
prolonged	O
hyperproliferation	O
of	O
hematopoietic	B-cell_type
cells	I-cell_type
,	O
but	O
rather	O
represents	O
a	O
primary	O
molecular	O
aberration	O
.	O

Constitutively	B-protein
active	I-protein
STAT3	I-protein
may	O
contribute	O
to	O
the	O
growth	O
factor	O
hypersensitivity	O
of	O
P.	B-cell_type
vera	I-cell_type
cells	I-cell_type
.	O

-DOCSTART-	O

Identification	O
of	O
phosphorylation	B-protein
sites	I-protein
for	O
Bruton	B-protein
's	I-protein
tyrosine	I-protein
kinase	I-protein
within	O
the	O
transcriptional	B-protein
regulator	I-protein
BAP/TFII-I	I-protein
.	O

Bruton	B-protein
's	I-protein
tyrosine	I-protein
kinase	I-protein
(	O
Btk	B-protein
)	O
,	O
a	O
member	O
of	O
the	O
Tec	B-protein
family	I-protein
of	I-protein
cytosolic	I-protein
kinases	I-protein
,	O
is	O
essential	O
for	O
B	B-cell_type
cell	I-cell_type
development	O
and	O
function	O
.	O

BAP/TFII-I	B-protein
,	O
a	O
protein	O
implicated	O
in	O
transcriptional	O
regulation	O
,	O
is	O
associated	O
with	O
Btk	B-protein
in	O
B	B-cell_type
cells	I-cell_type
and	O
is	O
transiently	O
phosphorylated	O
on	O
tyrosine	O
following	O
B	B-cell_type
cell	I-cell_type
receptor	O
engagement	O
.	O

BAP/TFII-I	B-protein
is	O
a	O
substrate	O
for	O
Btk	B-protein
in	O
vitro	O
and	O
is	O
hyperphosphorylated	O
on	O
tyrosine	O
upon	O
coexpression	O
with	O
Btk	B-protein
in	O
mammalian	B-cell_type
cells	I-cell_type
.	O

In	O
an	O
effort	O
to	O
understand	O
the	O
physiologic	O
consequences	O
of	O
BAP/TFII-I	B-protein
tyrosine	O
phosphorylation	O
following	O
B	O
cell	O
receptor	O
stimulation	O
,	O
site-directed	O
mutagenesis	O
and	O
phosphopeptide	O
mapping	O
were	O
used	O
to	O
locate	O
the	O
predominant	O
sites	O
of	O
BAP/TFII-I	B-protein
phosphorylation	O
by	O
Btk	B-protein
in	O
vitro	O
.	O

These	O
residues	O
,	O
Tyr248	O
,	O
Tyr357	O
,	O
and	O
Tyr462	O
,	O
were	O
also	O
found	O
to	O
be	O
the	O
major	O
sites	O
for	O
Btk	B-protein
-dependent	O
phosphorylation	O
of	O
BAP/TFII-I	B-protein
in	O
vivo	O
.	O

Residues	O
Tyr357	O
and	O
Tyr462	O
are	O
contained	O
within	O
the	O
loop	B-protein
regions	I-protein
of	O
adjacent	O
helix-loop-helix-like	B-protein
repeats	I-protein
within	O
BAP/TFII-I	B-protein
.	O

Mutation	O
of	O
either	O
Tyr248	O
,	O
Tyr357	O
,	O
or	O
Tyr462	O
to	O
phenylalanine	O
reduced	O
transcription	O
from	O
a	O
c-fos	B-DNA
promoter	I-DNA
relative	O
to	O
wild-type	B-DNA
BAP/TFII-I	I-DNA
in	O
transfected	O
COS-7	B-cell_line
cells	I-cell_line
,	O
consistent	O
with	O
the	O
interpretation	O
that	O
phosphorylation	O
at	O
these	O
sites	O
contributes	O
to	O
transcriptional	O
activation	O
.	O

Phosphorylation	O
of	O
BAP/TFII-I	B-protein
by	O
Btk	B-protein
may	O
link	O
engagement	O
of	O
receptors	O
such	O
as	O
surface	O
immunoglobulin	B-protein
to	O
modulation	O
of	O
gene	O
expression	O
.	O

-DOCSTART-	O

Expression	O
of	O
interferon	B-protein
consensus	I-protein
sequence	I-protein
binding	I-protein
protein	I-protein
induces	O
potent	O
immunity	O
against	O
BCR/ABL-induced	O
leukemia	O
.	O

Mice	O
deficient	O
in	O
the	O
interferon	B-protein
consensus	I-protein
sequence	I-protein
binding	I-protein
protein	I-protein
(	O
ICSBP	B-protein
)	O
develop	O
a	O
disease	O
resembling	O
chronic	O
myeloid	O
leukemia	O
(	O
CML	O
)	O
,	O
which	O
in	O
humans	O
is	O
caused	O
by	O
the	O
BCR/ABL	B-protein
oncoprotein	I-protein
.	O

Interferon-alpha	B-protein
(	O
IFN-alpha	B-protein
)	O
induces	O
ICSBP	B-protein
expression	O
and	O
is	O
an	O
effective	O
therapy	O
for	O
CML	O
.	O

This	O
study	O
examined	O
whether	O
enforced	O
expression	O
of	O
ICSBP	B-protein
might	O
antagonize	O
BCR/ABL-induced	O
leukemia	O
;	O
results	O
demonstrated	O
that	O
ICSBP-modified	B-cell_type
cells	I-cell_type
generated	O
a	O
protective	O
CD8	O
(	O
+	O
)	O
cytotoxic	O
T-cell	O
response	O
against	O
BCR/ABL-transformed	B-cell_type
BaF3	I-cell_type
cells	I-cell_type
in	O
a	O
murine	O
leukemia	O
model	O
.	O

ICSBP	B-protein
expression	O
represents	O
a	O
novel	O
means	O
of	O
stimulating	O
a	O
host	O
immune	O
response	O
to	O
BCR/ABL	B-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
leukemia	I-cell_type
cells	I-cell_type
and	O
a	O
potential	O
strategy	O
for	O
immunotherapy	O
of	O
CML	O
.	O

(	O
Blood.	O
2001	O
;	O
97	O
:	O
3491-3497	O
)	O

-DOCSTART-	O

A	O
transcriptional	O
block	O
in	O
the	O
IL-2	B-DNA
promoter	I-DNA
at	O
the	O
-150	B-DNA
AP-1	I-DNA
site	I-DNA
in	O
effector	B-cell_type
CD8+	I-cell_type
T	I-cell_type
cells	I-cell_type
.	O

Both	O
CD4+	B-cell_type
and	I-cell_type
CD8+	I-cell_type
T	I-cell_type
cells	I-cell_type
that	O
produce	O
IL-2	B-protein
in	O
response	O
to	O
Ag	O
recognition	O
have	O
been	O
isolated	O
.	O

However	O
,	O
most	O
effector	B-cell_type
CD8+	I-cell_type
T	I-cell_type
cells	I-cell_type
recovered	O
after	O
exposure	O
to	O
Ag	B-protein
do	O
not	O
produce	O
sufficient	O
IL-2	B-protein
to	O
sustain	O
growth	O
,	O
and	O
depend	O
on	O
CD4+	B-cell_type
T	I-cell_type
helper	I-cell_type
cells	I-cell_type
for	O
this	O
obligate	O
growth	O
factor	O
.	O

IL-2	O
expression	O
in	O
CD4+	B-cell_type
T	I-cell_type
cells	I-cell_type
is	O
primarily	O
controlled	O
at	O
the	O
level	O
of	O
transcription	O
,	O
but	O
mechanisms	O
restricting	O
IL-2	O
production	O
in	O
CD8+	B-cell_type
T	I-cell_type
cells	I-cell_type
have	O
not	O
been	O
elucidated	O
.	O

To	O
evaluate	O
transcriptional	O
regulation	O
of	O
the	O
IL-2	B-DNA
gene	I-DNA
in	O
CD8+	B-cell_type
T	I-cell_type
cells	I-cell_type
,	O
we	O
stably	O
transfected	O
reporter	B-DNA
genes	I-DNA
into	O
Ag	B-protein
-specific	O
CD8+	B-cell_line
T	I-cell_line
cell	I-cell_line
clones	I-cell_line
.	O

CD28+	B-cell_type
CD8	I-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
T	I-cell_type
cells	I-cell_type
unable	O
to	O
transcribe	O
the	O
IL-2	B-DNA
gene	I-DNA
in	O
response	O
to	O
antigenic	O
stimulation	O
had	O
a	O
block	O
in	O
transactivation	O
of	O
the	O
-150	B-DNA
CD28	I-DNA
response	I-DNA
element	I-DNA
(	I-DNA
CD28RE	I-DNA
)	I-DNA
/AP-1	I-DNA
site	I-DNA
of	O
the	O
IL-2	B-DNA
promoter	I-DNA
,	O
but	O
did	O
transactivate	O
the	O
composite	O
NFAT/AP-1	B-DNA
and	I-DNA
OCT/AP-1	I-DNA
sites	I-DNA
,	O
and	O
a	O
consensus	B-DNA
AP-1	I-DNA
motif	I-DNA
.	O

Mutation	O
of	O
the	O
nonconsensus	B-DNA
-150	I-DNA
AP-1	I-DNA
site	I-DNA
to	O
a	O
consensus	B-DNA
AP-1	I-DNA
site	I-DNA
,	O
or	O
insertion	O
of	O
a	O
CD28RE/AP-1	B-DNA
consensus	I-DNA
site	I-DNA
upstream	O
of	O
the	O
native	O
-150	B-DNA
CD28RE/AP-1	I-DNA
site	I-DNA
restored	O
transactivation	O
of	O
the	O
altered	O
promoter	B-DNA
.	O

These	O
results	O
suggest	O
that	O
the	O
defect	O
at	O
the	O
-150	B-DNA
site	I-DNA
may	O
reflect	O
the	O
absence	O
or	O
inactivity	O
of	O
a	O
required	O
factor	O
rather	O
than	O
repression	O
of	O
the	O
IL-2	B-DNA
promoter	I-DNA
.	O

-DOCSTART-	O

Stem	B-protein
cell	I-protein
factor	I-protein
and	O
interleukin-3	B-protein
induce	O
stepwise	O
generation	O
of	O
erythroid	B-cell_type
precursor	I-cell_type
cells	I-cell_type
from	O
a	O
basic	O
fibroblast	B-protein
growth	I-protein
factor	I-protein
-dependent	O
hematopoietic	B-cell_line
stem	I-cell_line
cell	I-cell_line
line	I-cell_line
,	I-cell_line
A-6	I-cell_line
.	O

A	O
m	O
ultipotent	B-cell_type
immature	I-cell_type
myeloid	I-cell_type
cell	I-cell_type
population	I-cell_type
was	O
produced	O
from	O
a	O
basic	B-protein
fibroblast	I-protein
growth	I-protein
factor	I-protein
(	O
bFGF	B-protein
)	O
-dependent	O
hematopoietic	B-cell_line
stem	I-cell_line
cell	I-cell_line
line	I-cell_line
,	I-cell_line
A-6	I-cell_line
,	O
when	O
cultured	O
with	O
stem	B-protein
cell	I-protein
factor	I-protein
(	O
SCF	B-protein
)	O
replacing	O
bFGF	B-protein
.	O

Those	O
cells	O
were	O
positive	O
for	O
stem	B-protein
cell	I-protein
markers	I-protein
,	O
c-kit	B-protein
and	O
CD34	B-protein
,	O
and	O
a	O
myeloid	B-protein
cell	I-protein
marker	I-protein
,	I-protein
F4/80	I-protein
.	O

Some	O
cell	O
fractions	O
were	O
also	O
positive	O
for	O
Mac-1	B-protein
,	I-protein
a	I-protein
macrophage	I-protein
marker	I-protein
or	O
Gr-1	B-protein
,	I-protein
a	I-protein
granulocytic	I-protein
maker	I-protein
,	O
but	O
negative	O
for	O
an	O
erythroid	B-protein
marker	I-protein
TER119	I-protein
.	O

They	O
also	O
showed	O
the	O
expression	O
of	O
mRNA	O
for	O
the	O
myeloid-specific	B-protein
PU.1	I-protein
but	O
did	O
not	O
that	O
for	O
the	O
erythroid-specific	B-protein
GATA-1	I-protein
.	O

Among	O
various	O
cytokines	B-protein
,	O
interleukin-3	B-protein
(	O
IL-3	B-protein
)	O
induced	O
erythroid	B-cell_type
precursor	I-cell_type
cells	I-cell_type
that	O
expressed	O
the	O
erythroid-specific	B-protein
GATA-1	I-protein
and	O
beta-major	B-protein
globin	I-protein
.	O

The	O
quantitative	O
analysis	O
showed	O
that	O
erythroid	B-cell_type
precursor	I-cell_type
cells	I-cell_type
were	O
newly	O
produced	O
from	O
the	O
immature	B-cell_type
myeloid	I-cell_type
cells	I-cell_type
by	O
cultivation	O
with	O
IL-3	B-protein
.	O

SCF	B-protein
and	O
IL-3	B-protein
induced	O
stepwise	O
generation	O
of	O
erythroid	B-cell_type
precursor	I-cell_type
cells	I-cell_type
from	O
an	O
A-6	B-cell_line
hematopoietic	I-cell_line
stem	I-cell_line
cell	I-cell_line
line	I-cell_line
.	O

Copyright	O
2001	O
Academic	O
Press	O
.	O

-DOCSTART-	O

Distinct	O
BMI-1	B-protein
and	O
EZH2	B-protein
expression	O
patterns	O
in	O
thymocytes	B-cell_type
and	O
mature	B-cell_type
T	I-cell_type
cells	I-cell_type
suggest	O
a	O
role	O
for	O
Polycomb	B-DNA
genes	I-DNA
in	O
human	O
T	O
cell	O
differentiation	O
.	O

BMI-1	B-protein
and	O
EZH2	B-protein
Polycomb-group	B-protein
(	I-protein
PcG	I-protein
)	I-protein
proteins	I-protein
belong	O
to	O
two	O
distinct	O
protein	O
complexes	O
involved	O
in	O
the	O
regulation	O
of	O
hematopoiesis	O
.	O

Using	O
unique	O
PcG-specific	O
antisera	O
and	O
triple	O
immunofluorescence	O
,	O
we	O
found	O
that	O
mature	B-cell_type
resting	I-cell_type
peripheral	I-cell_type
T	I-cell_type
cells	I-cell_type
expressed	O
BMI-1	B-protein
,	O
whereas	O
dividing	B-cell_type
blasts	I-cell_type
were	O
EZH2	O
(	O
+	O
)	O
.	O

By	O
contrast	O
,	O
subcapsular	B-cell_type
immature	I-cell_type
double-negative	I-cell_type
(	I-cell_type
DN	I-cell_type
)	I-cell_type
(	I-cell_type
CD4	I-cell_type
(	I-cell_type
-	I-cell_type
)	I-cell_type
/CD8	I-cell_type
(	I-cell_type
-	I-cell_type
)	I-cell_type
)	I-cell_type
T	I-cell_type
cells	I-cell_type
in	O
the	O
thymus	O
coexpressed	O
BMI-1	B-protein
and	O
EZH2	B-protein
or	O
were	O
BMI-1	O
single	O
positive	O
.	O

Their	O
descendants	O
,	O
double-positive	B-cell_type
(	I-cell_type
DP	I-cell_type
;	I-cell_type
CD4	I-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
/CD8	I-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
)	I-cell_type
cortical	I-cell_type
thymocytes	I-cell_type
,	O
expressed	O
EZH2	B-protein
without	O
BMI-1	B-protein
.	O

Most	O
EZH2	B-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
DN	I-cell_type
and	I-cell_type
DP	I-cell_type
thymocytes	I-cell_type
were	O
dividing	O
,	O
while	O
DN	B-cell_type
BMI-1	I-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
/EZH2	I-cell_type
(	I-cell_type
-	I-cell_type
)	I-cell_type
thymocytes	I-cell_type
were	O
resting	O
and	O
proliferation	O
was	O
occasionally	O
noted	O
in	O
DN	B-cell_type
BMI-1	I-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
/EZH2	I-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
cells	I-cell_type
.	O

Maturation	O
of	O
DP	B-cell_type
cortical	I-cell_type
thymocytes	I-cell_type
to	O
single-positive	B-cell_type
(	I-cell_type
CD4	I-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
/CD8	I-cell_type
(	I-cell_type
-	I-cell_type
)	I-cell_type
or	O
CD8	B-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
/CD4	I-cell_type
(	I-cell_type
-	I-cell_type
)	I-cell_type
)	I-cell_type
medullar	I-cell_type
thymocytes	I-cell_type
correlated	O
with	O
decreased	O
detectability	O
of	O
EZH2	B-protein
and	O
continued	O
relative	O
absence	O
of	O
BMI-1	B-protein
.	O

Our	O
data	O
show	O
that	O
BMI-1	B-protein
and	O
EZH2	B-protein
expression	O
in	O
mature	B-cell_type
peripheral	I-cell_type
T	I-cell_type
cells	I-cell_type
is	O
mutually	O
exclusive	O
and	O
linked	O
to	O
proliferation	O
status	O
,	O
and	O
that	O
this	O
pattern	O
is	O
not	O
yet	O
established	O
in	O
thymocytes	B-cell_type
of	O
the	O
cortex	O
and	O
medulla	O
.	O

T	B-cell_type
cell	I-cell_type
stage-specific	O
PcG	O
expression	O
profiles	O
suggest	O
that	O
PcG	B-DNA
genes	I-DNA
contribute	O
to	O
regulation	O
of	O
T	O
cell	O
differentiation	O
.	O

They	O
probably	O
reflect	O
stabilization	O
of	O
cell	O
type-specific	O
gene	O
expression	O
and	O
irreversibility	O
of	O
lineage	O
choice	O
.	O

The	O
difference	O
in	O
PcG	O
expression	O
between	O
medullar	B-cell_type
thymocytes	I-cell_type
and	O
mature	B-cell_type
interfollicular	I-cell_type
T	I-cell_type
cells	I-cell_type
indicates	O
that	O
additional	O
maturation	O
processes	O
occur	O
after	O
thymocyte	O
transportation	O
from	O
the	O
thymus	O
.	O

-DOCSTART-	O

Stepwise	O
lineage	O
restriction	O
of	O
progenitors	B-cell_type
in	O
lympho-myelopoiesis	O
.	O

It	O
has	O
long	O
been	O
controversial	O
whether	O
hematopoiesis	O
progresses	O
through	O
ordered	O
stages	O
of	O
determination	O
as	O
in	O
embryonic	O
development	O
.	O

This	O
is	O
due	O
to	O
the	O
absence	O
of	O
a	O
methodology	O
capable	O
of	O
exactly	O
determining	O
the	O
developmental	O
potential	O
of	O
hematopoietic	B-cell_type
stem/progenitor	I-cell_type
cells	I-cell_type
.	O

The	O
multilineage	O
progenitor	O
(	O
MLP	O
)	O
assay	O
enabled	O
us	O
to	O
discriminate	O
among	O
seven	O
types	O
of	O
hematopoietic	O
progenitors	O
,	O
which	O
are	O
multipotent	O
progenitor	O
p-MTB	O
(	O
capable	O
of	O
generating	O
myeloid	B-cell_type
,	I-cell_type
T	I-cell_type
and	I-cell_type
B	I-cell_type
cells	I-cell_type
)	O
,	O
bipotent	O
progenitors	O
p-MT	O
,	O
p-MB	O
and	O
p-TB	O
,	O
and	O
unipotent	O
progenitors	O
p-M	O
,	O
p-T	O
and	O
p-B	O
.	O

Among	O
these	O
seven	O
types	O
,	O
the	O
p-TB	O
type	O
progenitor	O
was	O
found	O
to	O
be	O
absent	O
.	O

These	O
findings	O
indicate	O
that	O
the	O
process	O
of	O
lineage	O
commitment	O
proceeds	O
through	O
an	O
ordered	O
but	O
not	O
random	O
process	O
.	O

By	O
extending	O
the	O
area	O
of	O
investigation	O
to	O
include	O
the	O
erythroid	B-cell_type
lineage	I-cell_type
,	O
more	O
convincing	O
evidence	O
for	O
the	O
ordered	O
process	O
was	O
obtained	O
.	O

Detailed	O
and	O
exact	O
illustration	O
of	O
the	O
process	O
of	O
hematopoiesis	O
will	O
provide	O
an	O
opportunity	O
to	O
revive	O
hematopoiesis	O
as	O
one	O
of	O
the	O
most	O
fascinating	O
targets	O
of	O
research	O
in	O
developmental	O
biology	O

-DOCSTART-	O

Epstein-Barr	O
Virus	O
and	O
its	O
glycoprotein-350	B-protein
upregulate	O
IL-6	B-protein
in	O
human	B-cell_type
B-lymphocytes	I-cell_type
via	O
CD21	B-protein
,	O
involving	O
activation	O
of	O
NF-kappaB	B-protein
and	O
different	O
signaling	O
pathways	O
.	O

Epstein-Barr	O
virus	O
(	O
EBV	O
)	O
is	O
a	O
ubiquitous	O
and	O
highly	O
immunotropic	O
gamma	O
herpesvirus	O
that	O
infects	O
more	O
than	O
90	O
%	O
of	O
humans	O
worldwide	O
.	O

Its	O
pathogenicity	O
leads	O
to	O
a	O
number	O
of	O
diseases	O
including	O
tumors	O
that	O
result	O
from	O
EBV	O
's	O
ability	O
to	O
readily	O
transform	O
B-lymphocytes	B-cell_type
and	O
,	O
to	O
a	O
lesser	O
extent	O
,	O
epithelial	B-cell_type
cells	I-cell_type
.	O

EBV	O
utilizes	O
CD21/CR2	B-protein
as	O
its	O
receptor	O
on	O
B	B-cell_type
cells	I-cell_type
to	O
initiate	O
the	O
infection	O
process	O
.	O

EBV	O
binds	O
to	O
CR2	B-protein
through	O
its	O
major	B-protein
envelope	I-protein
glycoprotein-350	I-protein
(	O
gp350	B-protein
)	O
and	O
is	O
also	O
a	O
remarkable	O
immunomodulating	O
agent	O
.	O

We	O
had	O
previously	O
shown	O
that	O
EBV	O
is	O
capable	O
of	O
modulating	O
the	O
synthesis	O
of	O
a	O
number	O
of	O
cytokines	B-protein
.	O

We	O
now	O
show	O
that	O
while	O
both	O
purified	O
recombinant	B-protein
gp350	I-protein
(	O
rgp350	B-protein
)	O
and	O
EBV	O
upregulate	O
IL-6	B-RNA
mRNA	I-RNA
synthesis	O
in	O
B	B-cell_type
cells	I-cell_type
,	O
EBV-induced	O
IL-6	O
gene	O
activation	O
occurs	O
for	O
a	O
significantly	O
longer	O
period	O
of	O
time	O
(	O
i.e.	O
12	O
hours	O
for	O
EBV	O
as	O
compared	O
to	O
6	O
hours	O
for	O
rgp350	B-protein
)	O
.	O

Moreover	O
,	O
the	O
half-life	O
of	O
EBV-induced	O
IL-6	B-RNA
mRNA	I-RNA
was	O
also	O
significantly	O
longer	O
(	O
10	O
hours	O
)	O
than	O
that	O
of	O
mRNA	O
induced	O
by	O
rgp350	B-protein
(	O
about	O
6	O
hours	O
)	O
.	O

Both	O
EBV	O
and	O
gp350	B-protein
enhance	O
the	O
binding	O
of	O
the	O
NF-kappaB	B-protein
transcription	I-protein
factor	I-protein
,	O
as	O
determined	O
by	O
band-shift	O
and	O
augment	O
NF-kappaB	B-protein
-mediated	O
activation	O
of	O
a	O
CAT	B-DNA
reporter	I-DNA
plasmid	I-DNA
.	O

Furthermore	O
,	O
we	O
demonstrate	O
that	O
while	O
the	O
activation	O
of	O
IL-6	O
gene	O
expression	O
by	O
gp350	B-protein
is	O
mediated	O
primarily	O
by	O
the	O
protein	O
kinase	O
C	O
pathway	O
,	O
EBV	O
can	O
mediate	O
its	O
effects	O
through	O
multiple	O
signaling	O
pathways	O
.	O

To	O
our	O
knowledge	O
this	O
is	O
the	O
first	O
report	O
showing	O
that	O
the	O
binding	O
of	O
a	O
herpesvirus	B-protein
envelope	I-protein
glycoprotein	I-protein
to	O
CR2	B-protein
on	O
human	B-cell_type
B	I-cell_type
cells	I-cell_type
results	O
in	O
the	O
activation	O
of	O
the	O
NF-kappaB	B-protein
transcription	I-protein
factor	I-protein
leading	O
to	O
the	O
upregulation	O
of	O
IL-6	O
gene	O
expression	O
in	O
these	O
lymphocytes	B-cell_type
.	O

Copyright	O
2001	O
Academic	O
Press	O
.	O

-DOCSTART-	O

Gene-	O
and	O
tissue-specificity	O
of	O
mutation	O
in	O
Big	O
Blue	O
rats	O
treated	O
with	O
the	O
hepatocarcinogen	O
N-hydroxy-2-acetylaminofluorene	O
.	O

In	O
a	O
previous	O
study	O
,	O
we	O
found	O
that	O
treating	O
transgenic	O
Big	O
Blue	O
rats	O
with	O
the	O
hepatocarcinogen	O
N-hydroxy-2-acetylaminofluorene	O
(	O
N-OH-AAF	O
)	O
produced	O
the	O
same	O
major	O
DNA	O
adduct	O
in	O
the	O
target	O
liver	O
and	O
the	O
nontarget	B-cell_type
spleen	I-cell_type
lymphocytes	I-cell_type
and	O
bone	B-cell_type
marrow	I-cell_type
cells	I-cell_type
,	O
induced	O
lacI	B-cell_type
mutants	I-cell_type
in	O
the	O
liver	O
,	O
and	O
induced	O
much	O
lower	O
frequencies	O
of	O
l	O
acI	B-cell_type
and	I-cell_type
hprt	I-cell_type
mutants	I-cell_type
in	O
spleen	B-cell_type
lymphocytes	I-cell_type
.	O

In	O
the	O
present	O
study	O
,	O
sequence	O
analysis	O
was	O
conducted	O
on	O
lacI	B-DNA
DNA	I-DNA
and	O
hprt	B-DNA
cDNA	I-DNA
from	O
the	O
mutants	O
,	O
to	O
determine	O
the	O
mutational	O
specificity	O
of	O
N-OH-AAF	O
in	O
the	O
rat	O
.	O

All	O
the	O
mutation	O
spectra	O
from	O
N-OH-AAF-treated	O
rats	O
differed	O
significantly	O
from	O
corresponding	O
mutation	O
profiles	O
from	O
untreated	O
animals	O
(	O
P	O
=	O
0.02	O
to	O
P	O
<	O
0.0001	O
)	O
.	O

Although	O
there	O
were	O
similarities	O
among	O
the	O
mutational	O
patterns	O
derived	O
from	O
N-OH-AAF-treated	O
rats	O
(	O
e.g.	O
,	O
G	O
:	O
C	O
--	O
>	O
T	O
:	O
A	O
transversion	O
was	O
the	O
most	O
common	O
mutation	O
in	O
all	O
mutation	O
sets	O
)	O
,	O
there	O
were	O
significant	O
differences	O
in	O
the	O
patterns	O
of	O
basepair	O
substitution	O
and	O
frameshift	O
mutation	O
between	O
the	O
liver	O
and	O
spleen	B-cell_type
lymphocyte	I-cell_type
lacI	I-cell_type
mutants	I-cell_type
(	O
P	O
=	O
0.02	O
)	O
and	O
between	O
the	O
spleen	B-cell_type
lymphocyte	I-cell_type
lacI	B-cell_type
and	I-cell_type
hprt	I-cell_type
mutants	I-cell_type
(	O
P	O
=	O
0.04	O
)	O
.	O

Also	O
,	O
multiplex	O
PCR	O
analysis	O
of	O
genomic	B-DNA
DNA	I-DNA
from	O
the	O
hprt	B-cell_type
mutants	I-cell_type
indicated	O
that	O
12	O
%	O
of	O
mutants	O
from	O
treated	O
rats	O
had	O
major	O
deletions	O
in	O
the	O
hprt	B-DNA
gene	I-DNA
;	O
no	O
corresponding	O
incidence	O
of	O
large	O
deletions	O
was	O
evident	O
among	O
lacI	O
mutations	O
.	O

All	O
the	O
mutation	O
profiles	O
reflect	O
the	O
general	O
mutational	O
specificity	O
of	O
the	O
major	O
DNA	O
adduct	O
formed	O
by	O
N-OH-AAF	O
.	O

The	O
differences	O
between	O
N-OH-AAF	O
mutation	O
in	O
the	O
endogenous	B-DNA
gene	I-DNA
and	O
transgene	B-DNA
can	O
be	O
partially	O
explained	O
by	O
the	O
structures	O
of	O
the	O
two	O
genes	O
.	O

The	O
tissue-specificity	O
of	O
the	O
mutation	O
spectra	O
may	O
contribute	O
to	O
targeting	O
tumor	O
formation	O
to	O
the	O
liver	O
.	O

Environ.	O
Mol.	O
Mutagen.	O
37	O
:	O
203-214	O
,	O
2001	O
.	O

Published	O
2001	O
Wiley-Liss	O
,	O
Inc	O
.	O

-DOCSTART-	O

Caspase	B-protein
-dependent	O
cleavage	O
of	O
the	O
hematopoietic	B-protein
specific	I-protein
adaptor	I-protein
protein	I-protein
Gads	I-protein
alters	O
signalling	O
from	O
the	O
T	B-protein
cell	I-protein
receptor	I-protein
.	O

Gads	B-protein
is	O
a	O
SH2	B-protein
and	I-protein
SH3	I-protein
domain	I-protein
-containing	O
,	O
hematopoietic-specific	B-protein
adaptor	I-protein
protein	I-protein
that	O
functions	O
in	O
signalling	O
from	O
the	O
T	B-protein
cell	I-protein
receptor	I-protein
.	O

Gads	B-protein
acts	O
by	O
linking	O
SLP-76	B-protein
,	O
bound	O
by	O
the	O
carboxy-terminal	B-protein
Gads	I-protein
SH3	I-protein
domain	I-protein
,	O
to	O
tyrosine	B-protein
phosphorylated	I-protein
LAT	I-protein
which	O
contains	O
binding	O
sites	O
for	O
the	O
Gads	B-protein
SH2	I-protein
domain	I-protein
.	O

Gads	B-protein
is	O
distinguished	O
from	O
Grb2	B-protein
and	O
the	O
closely	O
related	O
Grap	B-protein
protein	I-protein
by	O
the	O
presence	O
of	O
a	O
120	B-protein
amino	I-protein
acid	I-protein
unique	I-protein
region	I-protein
between	O
the	O
SH2	B-protein
domain	I-protein
and	O
the	O
carboxy	B-protein
terminal	I-protein
SH3	I-protein
domain	I-protein
.	O

Here	O
we	O
demonstrate	O
that	O
the	O
unique	O
region	O
of	O
Gads	B-protein
contains	O
a	O
capase	B-protein
cleavage	I-protein
site	I-protein
.	O

Induction	O
of	O
apoptosis	O
in	O
lymphocytes	B-cell_type
results	O
in	O
detectable	O
Gads	B-protein
cleavage	O
by	O
60	O
min	O
.	O

Gads	B-protein
cleavage	O
is	O
blocked	O
in	O
vivo	O
by	O
treating	O
cells	O
with	O
a	O
caspase	O
3	O
inhibitor	O
.	O

A	O
putative	B-protein
caspase	I-protein
3	I-protein
cleavage	I-protein
site	I-protein
was	O
identified	O
within	O
the	O
unique	O
region	O
and	O
mutation	O
of	O
this	O
site	O
prevented	O
Gads	B-protein
cleavage	O
in	O
vitro	O
,	O
and	O
in	O
vivo	O
.	O

The	O
Gads	B-protein
cleavage	O
products	O
retained	O
the	O
predicted	O
binding	O
specificity	O
for	O
SLP-76	B-protein
and	O
LAT	B-protein
.	O

Expression	O
of	O
the	O
Gads	B-protein
cleavage	O
products	O
in	O
Jurkat	B-cell_line
T	I-cell_line
cells	I-cell_line
inhibited	O
NFAT	O
activation	O
following	O
TCR	O
cross	O
linking	O
.	O

These	O
findings	O
indicate	O
that	O
cleavage	O
of	O
Gads	B-protein
in	O
vivo	O
could	O
function	O
to	O
alter	O
signalling	O
downstream	O
of	O
the	O
T	B-protein
cell	I-protein
receptor	I-protein
by	O
disrupting	O
cross	O
talk	O
between	O
SLP-76	B-protein
and	O
LAT	B-protein
.	O

-DOCSTART-	O

Targeting	O
of	O
p300	B-protein
to	O
the	O
interleukin-2	B-DNA
promoter	I-DNA
via	O
CREB-Rel	O
cross-talk	O
during	O
mitogen	B-protein
and	O
oncogenic	O
molecular	O
signaling	O
in	O
activated	B-cell_type
T-cells	I-cell_type
.	O

In	O
this	O
report	O
,	O
we	O
explore	O
the	O
mechanisms	O
of	O
targeting	O
of	O
p300	B-protein
to	O
the	O
interleukin-2	B-DNA
(	I-DNA
IL-2	I-DNA
)	I-DNA
promoter	I-DNA
in	O
response	O
to	O
mitogenic	O
and	O
oncogenic	O
molecular	O
signals	O
.	O

Recruitment	O
of	O
p300	B-protein
by	O
cAMP-responsive	B-protein
element-binding	I-protein
protein-Rel	I-protein
cross-talk	O
at	O
the	O
composite	B-protein
CD28	I-protein
response	I-protein
element	I-protein
(	O
CD28RE	B-protein
)	O
-	O
TRE	B-DNA
element	I-DNA
of	O
the	O
IL-2	B-DNA
promoter	I-DNA
is	O
essential	O
for	O
promoter	O
inducibility	O
during	O
T-cell	O
activation	O
,	O
and	O
CD28RE-TRE	B-DNA
is	O
the	O
exclusive	O
target	O
of	O
the	O
human	B-cell_type
T-cell	I-cell_type
lymphotropic	O
virus	O
type	O
I	O
oncoprotein	B-protein
Tax	I-protein
.	O

The	O
intrinsic	O
histone	O
acetyltransferase	O
activity	O
of	O
p300	B-protein
is	O
dispensable	O
for	O
activation	O
of	O
the	O
IL-2	B-DNA
promoter	I-DNA
,	O
and	O
the	O
N-terminal	B-protein
743	I-protein
residues	I-protein
contain	O
the	O
minimal	O
structural	O
requirements	O
for	O
synergistic	O
transactivation	O
of	O
the	O
CD28RE-TRE	B-DNA
,	O
the	O
IL-2	B-DNA
promoter	I-DNA
,	O
and	O
endogenous	O
IL-2	B-DNA
gene	I-DNA
expression	O
.	O

Mutational	O
analysis	O
of	O
p300	B-protein
reveals	O
differential	O
structural	O
requirements	O
for	O
the	O
N-terminal	O
p300	O
module	O
by	O
individual	O
cis-elements	B-DNA
within	O
the	O
IL-2	B-DNA
promoter	I-DNA
.	O

These	O
findings	O
provide	O
evidence	O
that	O
p300	B-protein
assembles	O
at	O
the	O
IL-2	B-DNA
promoter	I-DNA
to	O
form	O
an	O
enhanceosome-like	O
signal	O
transduction	O
target	O
that	O
is	O
centrally	O
integrated	O
at	O
the	O
CD28RE-TRE	B-DNA
element	I-DNA
of	O
the	O
IL-2	B-DNA
promoter	I-DNA
through	O
specific	O
protein	O
module-targeted	O
associations	O
in	O
activated	B-cell_type
T-cells	I-cell_type
.	O

-DOCSTART-	O

Regulation	O
of	O
cytokine	O
production	O
in	O
T-cell	B-cell_type
responses	O
to	O
inhalant	O
allergen	O
:	O
GATA-3	B-protein
expression	O
distinguishes	O
between	O
Th1-	O
and	O
Th2-polarized	O
immunity	O
.	O

BACKGROUND	O
:	O
The	O
precise	O
nature	O
of	O
allergen-specific	O
cytokine	O
responses	O
in	O
atopics	O
versus	O
non-atopics	O
,	O
in	O
particular	O
the	O
'Th1	O
polarity	O
'	O
of	O
responses	O
in	O
non-atopics	O
,	O
remains	O
controversial	O
.	O

This	O
is	O
due	O
in	O
part	O
to	O
the	O
relative	O
insensitivity	O
of	O
cytokine	O
detection	O
systems	O
,	O
and	O
associated	O
variations	O
in	O
kinetics	O
of	O
cytokine	O
production	O
and	O
catabolism	O
in	O
in	O
vitro	O
culture	O
systems	O
.	O

As	O
an	O
alternative	O
to	O
cytokine	O
measurement	O
,	O
this	O
study	O
focuses	O
on	O
expression	O
of	O
the	O
transcription	B-protein
factor	I-protein
GATA-3	I-protein
for	O
analysis	O
of	O
allergen-specific	O
Th	O
cell	O
responses	O
.	O

METHODS	O
:	O
Cord	B-cell_type
blood	I-cell_type
mononuclear	I-cell_type
cells	I-cell_type
were	O
Th1-	O
or	O
Th2-polarized	O
by	O
culture	O
in	O
IL-12-	O
or	O
IL-4-employing	O
established	O
methods	O
;	O
PBMC	B-cell_type
from	O
house	O
dust	O
mite	O
(	O
HDM	O
)	O
-sensitive	O
atopics	O
and	O
controls	O
were	O
stimulated	O
overnight	O
with	O
HDM	O
;	O
cytokine	O
production	O
was	O
measured	O
by	O
ELISA	O
and	O
GATA-3	B-RNA
mRNA	I-RNA
expression	O
by	O
PCR	O
.	O

RESULTS	O
:	O
Cytokine	B-protein
-driven	O
Th2	O
polarization	O
of	O
naive	B-cell_type
T	I-cell_type
cells	I-cell_type
is	O
associated	O
with	O
marked	O
upregulation	O
of	O
GATA-3	B-protein
expression	O
,	O
whereas	O
a	O
reciprocal	O
expression	O
pattern	O
accompanies	O
differentiation	O
towards	O
the	O
Th1	O
cytokine	O
phenotype	O
.	O

In	O
T	B-cell_type
cells	I-cell_type
from	O
HDM	O
skin	O
prick	O
test-positive	O
(	O
HDM-SPT+/HDM-IgE+	O
)	O
volunteers	O
,	O
overnight	O
stimulation	O
results	O
in	O
marked	O
upregulation	O
of	O
GATA-3	B-protein
expression	O
,	O
compared	O
to	O
an	O
equally	O
marked	O
downregulation	O
of	O
expression	O
in	O
T	B-cell_type
cells	I-cell_type
from	O
SPT-/IgE-	O
subjects	O
.	O

In	O
subjects	O
who	O
are	O
HDM-SPT+	O
but	O
IgE-	O
,	O
GATA-3	B-protein
expression	O
levels	O
remained	O
relatively	O
stable	O
during	O
culture	O
with	O
HDM	O
.	O

CONCLUSIONS	O
:	O
Upregulation	O
of	O
GATA-3	B-protein
expression	O
in	O
PBMC	B-cell_type
is	O
a	O
hallmark	O
of	O
the	O
early	O
phase	O
of	O
Th2	O
recall	O
responses	O
to	O
specific	O
allergen	O
in	O
atopics	O
.	O

The	O
reciprocal	O
expression	O
pattern	O
observed	O
in	O
HDM-specific	O
recall	O
responses	O
of	O
non-atopics	O
provides	O
independent	O
confirmation	O
of	O
the	O
presence	O
of	O
underlying	O
Th1-like	O
immunity	O
in	O
these	O
subjects	O
.	O

The	O
parallel	O
findings	O
in	O
neonatal	B-cell_type
T	I-cell_type
cells	I-cell_type
suggest	O
that	O
the	O
same	O
approach	O
may	O
be	O
utilized	O
for	O
monitoring	O
the	O
progress	O
of	O
allergen-specific	O
Th1/Th2	O
memory	O
development	O
during	O
early	O
childhood	O
,	O
and	O
hence	O
in	O
assessment	O
of	O
risk	O
for	O
future	O
allergic	O
disease	O
.	O

Copyright	O
2001	O
S.	O
Karger	O
AG	O
,	O
Basel	O

-DOCSTART-	O

The	O
heat	O
shock	O
response	O
reduces	O
myelin	B-protein
oligodendrocyte	I-protein
glycoprotein	I-protein
-induced	O
experimental	O
autoimmune	O
encephalomyelitis	O
in	O
mice	O
.	O

The	O
stress	O
response	O
(	O
SR	O
)	O
can	O
block	O
inflammatory	O
gene	O
expression	O
by	O
preventing	O
activation	O
of	O
transcription	B-protein
factor	I-protein
nuclear	B-protein
factor-kappa	I-protein
B	I-protein
(	O
NF-kappaB	B-protein
)	O
.	O

As	O
inflammatory	O
gene	O
expression	O
contributes	O
to	O
the	O
pathogenesis	O
of	O
demyelinating	O
diseases	O
,	O
we	O
tested	O
the	O
effects	O
of	O
the	O
SR	O
on	O
the	O
progression	O
of	O
the	O
demyelinating	O
disease	O
experimental	O
autoimmune	O
encephalomyelitis	O
(	O
EAE	O
)	O
.	O

EAE	O
was	O
actively	O
induced	O
in	O
C57BL/6	O
mice	O
using	O
an	O
encephalitogenic	O
myelin	B-protein
oligodendrocyte	I-protein
glycoprotein	I-protein
(	O
MOG	B-protein
(	I-protein
35-55	I-protein
)	I-protein
)	O
peptide	O
.	O

Whole	O
body	O
hyperthermia	O
was	O
used	O
to	O
induce	O
a	O
heat	O
shock	O
response	O
(	O
HSR	O
)	O
in	O
immunized	O
mice	O
2	O
days	O
after	O
the	O
booster	O
MOG	O
(	O
35-55	O
)	O
peptide	O
injection	O
.	O

The	O
HSR	O
reduced	O
the	O
incidence	O
of	O
EAE	O
by	O
70	O
%	O
,	O
delayed	O
disease	O
onset	O
by	O
6	O
days	O
,	O
and	O
attenuated	O
disease	O
severity	O
.	O

The	O
HSR	O
attenuated	O
leukocyte	B-cell_type
infiltration	O
into	O
CNS	O
assessed	O
by	O
quantitation	O
of	O
perivascular	O
infiltrates	O
,	O
and	O
by	O
reduced	O
staining	O
for	O
CD4	B-protein
and	O
CD25	B-protein
immunopositive	O
T-cells	B-cell_type
.	O

T-cell	O
activation	O
,	O
assessed	O
by	O
the	O
production	O
of	O
interferon	B-protein
gamma	I-protein
(	O
IFNgamma	B-protein
)	O
in	O
response	O
to	O
MOG	B-protein
(	I-protein
35-55	I-protein
)	I-protein
,	O
was	O
also	O
decreased	O
by	O
the	O
HSR	O
.	O

The	O
HSR	O
reduced	O
inflammatory	O
gene	O
expression	O
in	O
the	O
brain	O
that	O
normally	O
occurs	O
during	O
EAE	O
,	O
including	O
the	O
early	O
increase	O
in	O
RANTES	B-protein
(	O
regulated	B-protein
on	I-protein
activation	I-protein
of	I-protein
normal	I-protein
T-cell	I-protein
expressed	I-protein
and	I-protein
secreted	I-protein
)	O
expression	O
,	O
and	O
the	O
later	O
expression	O
of	O
the	O
inducible	O
form	O
of	O
nitric	B-protein
oxide	I-protein
synthase	I-protein
.	O

The	O
early	O
activation	O
of	O
transcription	B-protein
factor	I-protein
NF-kappaB	B-protein
was	O
also	O
blocked	O
by	O
the	O
HSR	O
.	O

The	O
finding	O
that	O
the	O
SR	O
reduces	O
inflammation	O
in	O
the	O
brain	O
and	O
the	O
clinical	O
severity	O
of	O
EAE	O
opens	O
a	O
novel	O
therapeutic	O
approach	O
for	O
prevention	O
of	O
autoimmune	O
diseases	O
.	O

-DOCSTART-	O

In	O
vivo	O
detection	O
of	O
intracellular	O
signaling	O
pathways	O
in	O
developing	O
thymocytes	B-cell_type
.	O

Information	O
regarding	O
the	O
intracellular	O
signaling	O
processes	O
that	O
occur	O
during	O
the	O
development	O
of	O
T	B-cell_type
cells	I-cell_type
has	O
largely	O
been	O
obtained	O
with	O
the	O
use	O
of	O
transgenic	O
mouse	O
models	O
,	O
which	O
although	O
providing	O
invaluable	O
information	O
are	O
time	O
consuming	O
and	O
costly	O
.	O

To	O
this	O
end	O
,	O
we	O
have	O
developed	O
a	O
novel	O
system	O
that	O
facilitates	O
the	O
in	O
vivo	O
analysis	O
of	O
signal	O
transduction	O
pathways	O
during	O
T-lymphocyte	B-cell_type
development	O
.	O

This	O
approach	O
uses	O
reporter-plasmids	B-DNA
for	O
the	O
detection	O
of	O
intracellular	O
signals	O
mediated	O
by	O
the	O
mitogen-activated	B-protein
protein	I-protein
kinase	I-protein
or	O
cyclic	B-protein
AMP-dependent	I-protein
protein	I-protein
kinase	I-protein
.	O

Reporter-plasmids	B-DNA
are	O
transfected	O
into	O
thymocytes	B-cell_type
in	O
fetal	O
thymic	O
organ	O
culture	O
by	O
accelerated	O
DNA/particle	O
bombardment	O
(	O
gene	O
gun	O
)	O
,	O
and	O
the	O
activation	O
of	O
a	O
signaling	O
pathway	O
is	O
determined	O
in	O
the	O
form	O
of	O
a	O
standard	O
luciferase	O
assay	O
.	O

Importantly	O
,	O
this	O
powerful	O
technique	O
preserves	O
the	O
structural	O
integrity	O
of	O
the	O
thymus	O
,	O
and	O
will	O
provide	O
an	O
invaluable	O
tool	O
to	O
study	O
how	O
thymocytes	B-cell_type
respond	O
to	O
normal	O
environmental	O
stimuli	O
encountered	O
during	O
differentiation	O
within	O
the	O
thymic	O
milieu	O
.	O

Thus	O
,	O
this	O
method	O
allows	O
for	O
the	O
monitoring	O
of	O
signals	O
that	O
occur	O
in	O
a	O
biological	O
time	O
frame	O
,	O
such	O
as	O
during	O
differentiation	O
,	O
and	O
within	O
the	O
natural	O
environment	O
of	O
differentiating	B-cell_type
cells	I-cell_type
.	O

-DOCSTART-	O

Core-binding	B-protein
factor	I-protein
beta	I-protein
(	O
CBFbeta	B-protein
)	O
,	O
but	O
not	O
CBFbeta-smooth	B-protein
muscle	I-protein
myosin	I-protein
heavy	I-protein
chain	I-protein
,	O
rescues	O
definitive	O
hematopoiesis	O
in	O
CBFbeta	B-protein
-deficient	O
embryonic	B-cell_type
stem	I-cell_type
cells	I-cell_type
.	O

Core-binding	B-protein
factor	I-protein
beta	I-protein
(	O
CBFbeta	B-protein
)	O
is	O
the	O
non-DNA-binding	B-protein
subunit	I-protein
of	O
the	O
heterodimeric	B-protein
CBFs	I-protein
.	O

Genes	B-DNA
encoding	I-DNA
CBFbeta	I-DNA
(	O
CBFB	B-DNA
)	O
,	O
and	O
one	O
of	O
the	O
DNA-binding	B-protein
CBFalpha	I-protein
subunits	I-protein
,	I-protein
Runx1	I-protein
(	O
also	O
known	O
as	O
CBFalpha2	B-protein
,	O
AML1	B-protein
,	O
and	O
PEBP2alphaB	B-protein
)	O
,	O
are	O
required	O
for	O
normal	O
hematopoiesis	O
and	O
are	O
also	O
frequent	O
targets	O
of	O
chromosomal	O
translocations	O
in	O
acute	O
leukemias	O
in	O
humans	O
.	O

Homozygous	O
disruption	O
of	O
either	O
the	O
Runx1	B-DNA
or	I-DNA
Cbfb	I-DNA
gene	I-DNA
in	O
mice	O
results	O
in	O
embryonic	O
lethality	O
at	O
midgestation	O
due	O
to	O
hemorrhaging	O
in	O
the	O
central	O
nervous	O
system	O
,	O
and	O
severely	O
impairs	O
fetal	O
liver	O
hematopoiesis	O
.	O

Results	O
of	O
this	O
study	O
show	O
that	O
Cbfb-deficient	O
mouse	O
embryonic	B-cell_type
stem	I-cell_type
(	I-cell_type
ES	I-cell_type
)	I-cell_type
cells	I-cell_type
can	O
differentiate	O
into	O
primitive	B-cell_type
erythroid	I-cell_type
colonies	I-cell_type
in	O
vitro	O
,	O
but	O
are	O
impaired	O
in	O
their	O
ability	O
to	O
produce	O
definitive	O
erythroid	B-cell_type
and	I-cell_type
myeloid	I-cell_type
colonies	I-cell_type
,	O
mimicking	O
the	O
in	O
vivo	O
defect	O
.	O

Definitive	O
hematopoiesis	O
is	O
restored	O
by	O
ectopic	O
expression	O
of	O
full-length	B-DNA
Cbfb	I-DNA
transgenes	I-DNA
,	O
as	O
well	O
as	O
by	O
a	O
transgene	O
encoding	O
only	O
the	O
heterodimerization	B-protein
domain	I-protein
of	O
CBFbeta	B-protein
.	O

In	O
contrast	O
,	O
the	O
CBFbeta	B-protein
-	O
smooth	B-protein
muscle	I-protein
myosin	I-protein
heavy	I-protein
chain	I-protein
(	I-protein
SMMHC	I-protein
)	I-protein
fusion	I-protein
protein	I-protein
generated	O
by	O
the	O
inv	B-protein
(	I-protein
16	I-protein
)	I-protein
associated	O
with	O
acute	B-cell_line
myeloid	I-cell_line
leukemias	I-cell_line
(	O
M4Eo	B-cell_line
)	O
can	O
not	O
rescue	O
definitive	O
hematopoiesis	O
by	O
Cbfb-deficient	B-cell_type
ES	I-cell_type
cells	I-cell_type
.	O

Sequences	O
responsible	O
for	O
the	O
inability	O
of	O
CBFbeta	B-protein
-SMMHC	O
to	O
rescue	O
definitive	O
hematopoiesis	O
reside	O
in	O
the	O
SMMHC	B-protein
portion	I-protein
of	O
the	O
fusion	B-protein
protein	I-protein
.	O

Results	O
also	O
show	O
that	O
the	O
CBFbeta-SMMHC	B-protein
fusion	I-protein
protein	I-protein
transdominantly	O
inhibits	O
definitive	O
hematopoiesis	O
,	O
but	O
not	O
to	O
the	O
same	O
extent	O
as	O
homozygous	O
loss	O
of	O
Runx1	B-protein
or	O
Cbfb	B-protein
.	O

CBFbeta-SMMHC	B-protein
preferentially	O
inhibits	O
the	O
differentiation	O
of	O
myeloid	B-cell_type
lineage	I-cell_type
cells	I-cell_type
,	O
while	O
increasing	O
the	O
number	O
of	O
blastlike	B-cell_type
cells	I-cell_type
in	O
culture	O
.	O

-DOCSTART-	O

The	O
latency	O
pattern	O
of	O
Epstein-Barr	O
virus	O
infection	O
and	O
viral	O
IL-10	B-protein
expression	O
in	O
cutaneous	B-cell_type
natural	I-cell_type
killer/T-cell	I-cell_type
lymphomas	I-cell_type
.	O

The	O
nasal	B-cell_type
type	I-cell_type
,	I-cell_type
extranodal	I-cell_type
natural	I-cell_type
killer	I-cell_type
or	I-cell_type
T	I-cell_type
(	I-cell_type
NK/T	I-cell_type
)	I-cell_type
-cell	I-cell_type
lymphoma	I-cell_type
is	O
usually	O
associated	O
with	O
latent	O
Epstein-Barr	O
virus	O
(	O
EBV	O
)	O
infection	O
.	O

In	O
order	O
to	O
elucidate	O
the	O
EBV	O
gene	O
expression	O
patterns	O
in	O
vivo	O
,	O
we	O
examined	O
eight	O
patients	O
with	O
cutaneous	O
EBV-related	O
NK/T-cell	O
lymphomas	O
,	O
including	O
six	O
patients	O
with	O
a	O
NK-cell	O
phenotype	O
and	O
two	O
patients	O
with	O
a	O
T-cell	O
phenotype	O
.	O

The	O
implication	O
of	O
EBV	O
in	O
the	O
skin	O
lesions	O
was	O
determined	O
by	O
the	O
presence	O
of	O
EBV-DNA	B-DNA
,	O
EBV-encoded	B-RNA
nuclear	I-RNA
RNA	I-RNA
(	O
EBER	B-RNA
)	O
and	O
a	O
clonality	O
of	O
EBV-DNA	B-DNA
fragments	I-DNA
containing	O
the	O
terminal	B-DNA
repeats	I-DNA
.	O

Transcripts	O
of	O
EBV-encoded	B-DNA
genes	I-DNA
were	O
screened	O
by	O
reverse	O
transcription-	O
polymerase	O
chain	O
reaction	O
(	O
RT-PCR	O
)	O
,	O
and	O
confirmed	O
by	O
Southern	O
blot	O
hybridization	O
.	O

The	O
expression	O
of	O
EBV-related	B-protein
antigens	I-protein
was	O
examined	O
by	O
immunostaining	O
using	O
paraffin-embedded	O
tissue	O
sections	O
and	O
cell	O
pellets	O
of	O
EBV-positive	B-cell_line
cell	I-cell_line
lines	I-cell_line
.	O

Our	O
study	O
demonstrated	O
that	O
all	O
samples	O
from	O
the	O
patients	O
contained	O
EBV	B-RNA
nuclear	I-RNA
antigen	I-RNA
(	I-RNA
EBNA	I-RNA
)	I-RNA
-1	I-RNA
mRNA	I-RNA
which	O
was	O
transcribed	O
using	O
the	O
Q	B-DNA
promoter	I-DNA
,	O
whereas	O
both	O
the	O
Q	B-DNA
promoter	I-DNA
and	O
another	O
upstream	B-DNA
promoter	I-DNA
(	O
Cp/Wp	B-DNA
)	O
were	O
used	O
in	O
EBV-positive	B-cell_line
cell	I-cell_line
lines	I-cell_line
,	O
B95.8	B-cell_line
,	O
Raji	B-cell_line
and	O
Jiyoye	B-cell_line
.	O

Latent	B-RNA
membrane	I-RNA
protein-1	I-RNA
(	I-RNA
LMP-1	I-RNA
)	I-RNA
mRNA	I-RNA
was	O
detected	O
in	O
seven	O
of	O
eight	O
patients	O
and	O
all	O
cell	B-cell_line
lines	I-cell_line
,	O
whereas	O
EBNA-2	B-RNA
transcripts	I-RNA
were	O
found	O
only	O
in	O
the	O
cell	B-cell_line
lines	I-cell_line
.	O

Immunostaining	O
showed	O
no	O
LMP-1	B-protein
,	I-protein
EBNA-2	I-protein
or	I-protein
ZEBRA	I-protein
antigens	I-protein
in	O
the	O
paraffin-embedded	O
tissue	O
sections	O
,	O
although	O
they	O
were	O
positive	O
in	O
the	O
cell	B-cell_line
line	I-cell_line
cells	I-cell_line
.	O

Latent	B-RNA
BHRF1	I-RNA
transcripts	I-RNA
encoding	O
bcl-2	B-RNA
homologue	I-RNA
and	O
BCRF1	B-RNA
transcripts	I-RNA
encoding	O
viral	B-protein
interleukin	I-protein
(	I-protein
vIL	I-protein
)	I-protein
-10	I-protein
were	O
detected	O
in	O
one	O
and	O
two	O
of	O
eight	O
patients	O
,	O
respectively	O
.	O

A	O
patient	O
with	O
NK-cell	O
lymphoma	O
expressing	O
both	O
transcripts	O
died	O
of	O
rapid	O
progression	O
of	O
the	O
illness	O
.	O

Our	O
results	O
indicate	O
that	O
the	O
restricted	O
expression	O
of	O
the	O
latency-associated	B-DNA
EBV	I-DNA
genes	I-DNA
and	O
the	O
production	O
of	O
vIL-10	B-RNA
and	I-RNA
bcl-2	I-RNA
homologue	I-RNA
may	O
favour	O
tumour	O
growth	O
,	O
evading	O
the	O
host	O
immune	O
surveillance	O
.	O

Copyright	O
2001	O
Cancer	O
Research	O
Campaign	O
.	O

-DOCSTART-	O

Oxidized	O
alkyl	O
phospholipids	O
are	O
specific	O
,	O
high	O
affinity	O
peroxisome	O
proliferator-activated	O
receptor	O
gamma	O
ligands	O
and	O
agonists	O
.	O

Synthetic	O
high	O
affinity	O
peroxisome	O
proliferator-activated	O
receptor	O
(	O
PPAR	O
)	O
agonists	O
are	O
known	O
,	O
but	O
biologic	O
ligands	O
are	O
of	O
low	O
affinity	O
.	O

Oxidized	B-protein
low	I-protein
density	I-protein
lipoprotein	I-protein
(	O
oxLDL	B-protein
)	O
is	O
inflammatory	O
and	O
signals	O
through	O
PPARs	B-protein
.	O

We	O
showed	O
,	O
by	O
phospholipase	O
A	O
(	O
1	O
)	O
digestion	O
,	O
that	O
PPARgamma	O
agonists	O
in	O
oxLDL	B-protein
arise	O
from	O
the	O
small	O
pool	O
of	O
alkyl	O
phosphatidylcholines	O
in	O
LDL	B-protein
.	O

We	O
identified	O
an	O
abundant	O
oxidatively	O
fragmented	O
alkyl	O
phospholipid	O
in	O
oxLDL	B-protein
,	O
hexadecyl	O
azelaoyl	O
phosphatidylcholine	O
(	O
azPC	O
)	O
,	O
as	O
a	O
high	O
affinity	O
ligand	O
and	O
agonist	O
for	O
PPARgamma	B-protein
.	O

[	O
(	O
3	O
)	O
H	O
]	O
azPC	O
bound	O
recombinant	B-protein
PPARgamma	I-protein
with	O
an	O
affinity	O
(	O
K	O
(	O
d	O
)	O
(	O
(	O
app	O
)	O
)	O
approximately	O
40	O
nm	O
)	O
that	O
was	O
equivalent	O
to	O
rosiglitazone	O
(	O
BRL49653	O
)	O
,	O
and	O
competition	O
with	O
rosiglitazone	O
showed	O
that	O
binding	O
occurred	O
in	O
the	O
ligand-binding	B-protein
pocket	I-protein
.	O

azPC	O
induced	O
PPRE	O
reporter	O
gene	O
expression	O
,	O
as	O
did	O
rosiglitazone	O
,	O
with	O
a	O
half-maximal	O
effect	O
at	O
100	O
nm	O
.	O

Overexpression	O
of	O
PPARalpha	B-protein
or	O
PPARgamma	B-protein
revealed	O
that	O
azPC	O
was	O
a	O
specific	O
PPARgamma	O
agonist	O
.	O

The	O
scavenger	B-protein
receptor	I-protein
CD36	I-protein
is	O
encoded	O
by	O
a	O
PPRE-responsive	B-DNA
gene	I-DNA
,	O
and	O
azPC	O
enhanced	O
expression	O
of	O
CD36	B-protein
in	O
primary	B-cell_type
human	I-cell_type
monocytes	I-cell_type
.	O

We	O
found	O
that	O
anti-CD36	B-protein
inhibited	O
azPC	O
uptake	O
,	O
and	O
it	O
inhibited	O
PPRE	O
reporter	O
induction	O
.	O

Results	O
with	O
a	O
small	O
molecule	O
phospholipid	B-protein
flippase	I-protein
mimetic	I-protein
suggest	O
azPC	O
acts	O
intracellularly	O
and	O
that	O
cellular	O
azPC	O
accumulation	O
was	O
efficient	O
.	O

Thus	O
,	O
certain	O
alkyl	O
phospholipid	O
oxidation	O
products	O
in	O
oxLDL	B-protein
are	O
specific	O
,	O
high	O
affinity	O
extracellular	O
ligands	O
and	O
agonists	O
for	O
PPARgamma	B-protein
that	O
induce	O
PPAR-responsive	B-DNA
genes	I-DNA

-DOCSTART-	O

Regulation	O
of	O
the	O
human	B-DNA
MAT2B	I-DNA
gene	I-DNA
encoding	O
the	O
regulatory	B-protein
beta	I-protein
subunit	I-protein
of	O
methionine	B-protein
adenosyltransferase	I-protein
,	O
MAT	B-protein
II	I-protein
.	O

Methionine	B-protein
adenosyltransferase	I-protein
(	O
MAT	B-protein
)	O
catalyzes	O
the	O
biosynthesis	O
of	O
S-adenosylmethionine	B-protein
(	O
AdoMet	B-protein
)	O
,	O
a	O
key	O
molecule	O
in	O
transmethylation	O
reactions	O
and	O
polyamine	O
biosynthesis	O
.	O

The	O
MAT	B-protein
II	I-protein
isozyme	I-protein
consists	O
of	O
a	O
catalytic	B-protein
alpha2	I-protein
and	I-protein
a	I-protein
regulatory	I-protein
beta	I-protein
subunit	I-protein
.	O

Down-regulation	O
of	O
the	O
MAT	B-protein
II	I-protein
beta	I-protein
subunit	I-protein
expression	O
causes	O
a	O
6-10-fold	O
increase	O
in	O
intracellular	O
AdoMet	B-protein
levels	O
.	O

To	O
understand	O
the	O
mechanism	O
by	O
which	O
the	O
beta	B-protein
subunit	I-protein
expression	O
is	O
regulated	O
,	O
we	O
cloned	O
the	O
MAT2B	B-DNA
gene	I-DNA
,	O
determined	O
its	O
organization	O
,	O
characterized	O
its	O
5'-flanking	B-DNA
sequences	I-DNA
,	O
and	O
elucidated	O
the	O
in	O
vitro	O
and	O
in	O
vivo	O
regulation	O
of	O
its	O
promoter	B-DNA
.	O

Transcription	O
of	O
the	O
MAT2B	B-DNA
gene	I-DNA
initiates	O
at	O
position	O
-203	O
relative	O
to	O
the	O
translation	B-DNA
start	I-DNA
site	I-DNA
.	O

Promoter	B-DNA
deletion	I-DNA
analysis	I-DNA
defined	O
a	O
minimal	B-DNA
promoter	I-DNA
between	O
positions	B-DNA
+52	I-DNA
and	I-DNA
+93	I-DNA
base	O
pairs	O
,	O
a	O
GC-rich	B-DNA
region	I-DNA
.	O

Inclusion	O
of	O
the	O
sequences	O
between	O
-4	O
and	O
+52	O
enhanced	O
promoter	O
activity	O
;	O
this	O
was	O
primarily	O
because	O
of	O
an	O
Sp1	B-DNA
recognition	I-DNA
site	I-DNA
at	O
+9/+15	O
.	O

The	O
inclusion	O
of	O
sequences	O
up	O
to	O
position	O
-115	O
provided	O
full	O
activity	O
;	O
this	O
was	O
attributed	O
to	O
a	O
TATA	B-DNA
at	O
-32	O
.	O

The	O
Sp1	B-DNA
site	I-DNA
at	O
position	O
+9	O
was	O
key	O
for	O
the	O
formation	O
of	O
protein.DNA	O
complexes	O
.	O

Mutation	O
of	O
both	O
the	O
Sp1	B-DNA
site	I-DNA
at	O
+9	O
and	O
the	O
TATA	B-DNA
at	O
-32	O
reduced	O
promoter	O
activity	O
to	O
its	O
minimal	O
level	O
.	O

Supershift	O
assays	O
showed	O
no	O
effect	O
of	O
the	O
anti-Sp1	B-protein
antibody	I-protein
on	O
complex	O
formation	O
,	O
whereas	O
the	O
anti-Sp3	B-protein
antibody	I-protein
had	O
a	O
strong	O
effect	O
on	O
protein.DNA	O
complex	O
formation	O
,	O
suggesting	O
that	O
Sp3	B-protein
is	O
one	O
of	O
the	O
main	O
factors	O
binding	O
to	O
this	O
Sp1	B-DNA
site	I-DNA
.	O

Chromatin	O
immunoprecipitation	O
assays	O
supported	O
the	O
involvement	O
of	O
both	O
Sp1	B-protein
and	O
Sp3	B-protein
in	O
complexes	O
formed	O
on	O
the	O
MAT2B	B-DNA
promoter	I-DNA
.	O

The	O
data	O
show	O
that	O
the	O
5'-untranslated	B-DNA
sequences	I-DNA
play	O
an	O
important	O
role	O
in	O
regulating	O
the	O
MAT2B	B-DNA
gene	I-DNA
and	O
identifies	O
the	O
Sp1	B-DNA
site	I-DNA
at	O
+9	O
as	O
a	O
potential	O
target	O
for	O
modulating	O
MAT2B	O
expression	O
,	O
a	O
process	O
that	O
can	O
have	O
a	O
major	O
effect	O
on	O
intracellular	O
AdoMet	B-protein
levels	O
.	O

-DOCSTART-	O

Molecular	O
pathogenesis	O
of	O
influenza	O
A	O
virus	O
infection	O
and	O
virus-induced	O
regulation	O
of	O
cytokine	B-DNA
gene	I-DNA
expression	O
.	O

Despite	O
vaccines	O
and	O
antiviral	O
substances	O
influenza	O
still	O
causes	O
significant	O
morbidity	O
and	O
mortality	O
world	O
wide	O
.	O

Better	O
understanding	O
of	O
the	O
molecular	O
mechanisms	O
of	O
influenza	O
virus	O
replication	O
,	O
pathogenesis	O
and	O
host	O
immune	O
responses	O
is	O
required	O
for	O
the	O
development	O
of	O
more	O
efficient	O
means	O
of	O
prevention	O
and	O
treatment	O
of	O
influenza	O
.	O

Influenza	O
A	O
virus	O
,	O
which	O
replicates	O
in	O
epithelial	B-cell_type
cells	I-cell_type
and	O
leukocytes	B-cell_type
,	O
regulates	O
host	O
cell	O
transcriptional	O
and	O
translational	O
systems	O
and	O
activates	O
,	O
as	O
well	O
as	O
downregulates	O
apoptotic	O
pathways	O
.	O

Influenza	O
A	O
virus	O
infection	O
results	O
in	O
the	O
production	O
of	O
chemotactic	O
(	O
RANTES	B-protein
,	O
MIP-1	B-protein
alpha	I-protein
,	O
MCP-1	B-protein
,	O
MCP-3	B-protein
,	O
and	O
IP-10	B-protein
)	O
,	O
pro-inflammatory	O
(	O
IL-1	B-protein
beta	I-protein
,	O
IL-6	B-protein
,	O
IL-18	B-protein
,	O
and	O
TNF-alpha	B-protein
)	O
,	O
and	O
antiviral	B-protein
(	I-protein
IFN-alpha/beta	I-protein
)	I-protein
cytokines	I-protein
.	O

Cytokine	O
gene	O
expression	O
is	O
associated	O
with	O
the	O
activation	O
of	O
NF-kappa	B-protein
B	I-protein
,	O
AP-1	B-protein
,	O
STAT	B-protein
and	O
IRF	B-protein
signal	I-protein
transducing	I-protein
molecules	I-protein
in	O
influenza	O
A	O
virus-infected	O
cells	O
.	O

In	O
addition	O
of	O
upregulating	O
cytokine	B-DNA
gene	I-DNA
expression	O
,	O
influenza	O
A	O
virus	O
infection	O
activates	O
caspase-1	B-protein
enzyme	I-protein
,	O
which	O
is	O
involved	O
in	O
the	O
proteolytic	O
processing	O
of	O
proIL-1	B-protein
beta	I-protein
and	O
proIL-18	B-protein
into	O
their	O
biologically	O
active	O
forms	O
.	O

Influenza	O
A	O
virus-induced	O
IFN-alpha/beta	B-protein
is	O
essential	O
in	O
host	O
's	O
antiviral	O
defence	O
by	O
activating	O
the	O
expression	O
of	O
antiviral	B-DNA
Mx	I-DNA
,	O
PKR	B-DNA
and	I-DNA
oligoadenylate	I-DNA
synthetase	I-DNA
genes	I-DNA
.	O

IFN-alpha/beta	B-protein
also	O
prolongs	O
T	O
cell	O
survival	O
,	O
upregulates	O
IL-12	O
and	O
IL-18	O
receptor	O
gene	O
expression	O
and	O
together	O
with	O
IL-18	B-protein
stimulates	O
NK	B-cell_type
and	I-cell_type
T	I-cell_type
cell	I-cell_type
IFN-gamma	O
production	O
and	O
the	O
development	O
of	O
Th1-type	O
immune	O
response	O
.	O

-DOCSTART-	O

Comparison	O
of	O
hprt	O
and	O
lacI	O
mutant	O
frequency	O
with	O
DNA	O
adduct	O
formation	O
in	O
N-hydroxy-2-acetylaminofluorene-treated	O
Big	O
Blue	O
rats	O
.	O

N-Hydroxy-2-acetylaminofluorene	O
(	O
N-OH-AAF	O
)	O
is	O
the	O
proximate	O
carcinogenic	O
metabolite	O
of	O
the	O
powerful	O
rat	O
liver	O
carcinogen	O
2-acetylaminofluorene	O
.	O

In	O
this	O
study	O
,	O
transgenic	O
Big	O
Blue	O
(	O
R	O
)	O
rats	O
were	O
used	O
to	O
examine	O
the	O
relationship	O
between	O
in	O
vivo	O
mutagenicity	O
and	O
DNA	O
adduct	O
formation	O
by	O
N-OH-AAF	O
in	O
the	O
target	O
liver	O
compared	O
with	O
that	O
in	O
nontarget	O
tissues	O
.	O

Male	O
rats	O
were	O
given	O
one	O
,	O
two	O
,	O
or	O
four	O
doses	O
of	O
25	O
mg	O
N-OH-AAF/kg	O
body	O
weight	O
by	O
i.p.	O
injection	O
at	O
4-day	O
intervals	O
,	O
and	O
groups	O
of	O
treated	O
and	O
control	O
rats	O
were	O
euthanized	O
up	O
to	O
10	O
weeks	O
after	O
beginning	O
the	O
dosing	O
.	O

Mutant	O
frequencies	O
were	O
measured	O
in	O
the	O
spleen	B-DNA
lymphocyte	I-DNA
hprt	I-DNA
gene	I-DNA
,	O
and	O
lacI	O
mutant	O
frequencies	O
were	O
determined	O
in	O
the	O
liver	O
and	O
spleen	O
lymphocytes	B-cell_type
.	O

At	O
6	O
weeks	O
after	O
beginning	O
the	O
dosing	O
,	O
the	O
hprt	O
mutant	O
frequency	O
in	O
spleen	B-cell_type
lymphocytes	I-cell_type
from	O
the	O
four-dose	O
group	O
was	O
16.5	O
x	O
10	O
(	O
-6	O
)	O
compared	O
with	O
3.2	O
x	O
10	O
(	O
-6	O
)	O
in	O
control	O
animals	O
.	O

Also	O
at	O
6	O
weeks	O
,	O
rats	O
given	O
one	O
,	O
two	O
,	O
or	O
four	O
doses	O
of	O
N-OH-AAF	O
had	O
lacI	O
mutant	O
frequencies	O
in	O
the	O
liver	O
of	O
97.6	O
,	O
155.6	O
,	O
and	O
406.8	O
x	O
10	O
(	O
-6	O
)	O
,	O
respectively	O
,	O
compared	O
with	O
a	O
control	O
frequency	O
of	O
25.7	O
x	O
10	O
(	O
-6	O
)	O
;	O
rats	O
given	O
four	O
doses	O
had	O
lacI	O
mutant	O
frequencies	O
in	O
spleen	B-cell_type
lymphocytes	I-cell_type
of	O
55.8	O
x	O
10	O
(	O
-6	O
)	O
compared	O
with	O
a	O
control	O
frequency	O
of	O
20.4	O
x	O
10	O
(	O
-6	O
)	O
.	O

Additional	O
rats	O
were	O
evaluated	O
for	O
DNA	O
adduct	O
formation	O
in	O
the	O
liver	O
,	O
spleen	B-cell_type
lymphocytes	I-cell_type
,	O
and	O
bone	O
marrow	O
by	O
(	O
32	O
)	O
P-postlabeling	O
.	O

Adduct	O
analysis	O
was	O
conducted	O
1	O
day	O
after	O
one	O
,	O
two	O
,	O
and	O
four	O
treatments	O
with	O
N-OH-AAF	O
,	O
5	O
days	O
after	O
one	O
treatment	O
,	O
and	O
9	O
days	O
after	O
two	O
treatments	O
.	O

N-	O
(	O
Deoxyguanosin-8-yl	O
)	O
-2-aminofluorene	O
was	O
the	O
major	O
DNA	O
adduct	O
identified	O
in	O
all	O
the	O
tissues	O
examined	O
.	O

Adduct	O
concentrations	O
increased	O
with	O
total	O
dose	O
to	O
maximum	O
values	O
in	O
samples	O
taken	O
1	O
day	O
after	O
two	O
doses	O
,	O
and	O
remained	O
essentially	O
the	O
same	O
after	O
four	O
doses	O
.	O

In	O
samples	O
taken	O
after	O
four	O
doses	O
,	O
adduct	O
levels	O
were	O
103	O
,	O
28	O
,	O
and	O
7	O
fmol/microg	O
of	O
DNA	O
in	O
liver	O
,	O
spleen	B-cell_type
lymphocytes	I-cell_type
,	O
and	O
bone	O
marrow	O
,	O
respectively	O
.	O

The	O
results	O
indicate	O
that	O
the	O
extent	O
of	O
both	O
DNA	O
adduct	O
formation	O
and	O
mutant	O
induction	O
correlates	O
with	O
the	O
organ	O
specificity	O
for	O
N-OH-AAF	O
carcinogenesis	O
in	O
the	O
rat	O
.	O

Environ.	O
Mol.	O
Mutagen.	O
37	O
:	O
195-202	O
,	O
2001	O
.	O

Published	O
2001	O
Wiley-Liss	O
,	O
Inc	O
.	O

-DOCSTART-	O

Expression	O
of	O
Mad1	B-protein
in	O
T	B-cell_type
cells	I-cell_type
leads	O
to	O
reduced	O
thymic	O
cellularity	O
and	O
impaired	O
mitogen-induced	O
proliferation	O
.	O

To	O
investigate	O
Mad1	B-protein
function	O
in	O
vivo	O
,	O
transgenic	O
mice	O
were	O
generated	O
that	O
express	O
a	O
Mad1	B-DNA
transgene	I-DNA
in	O
T	B-cell_type
lineage	I-cell_type
cells	I-cell_type
under	O
the	O
control	O
of	O
the	O
proximal	B-DNA
lck	I-DNA
promoter	I-DNA
.	O

Thymus	O
size	O
in	O
lck-Mad1	O
transgenic	O
mice	O
is	O
drastically	O
reduced	O
although	O
representation	O
of	O
the	O
various	O
thymocyte	B-cell_type
sub	I-cell_type
populations	I-cell_type
appears	O
normal	O
.	O

To	O
investigate	O
more	O
closely	O
any	O
effects	O
of	O
Mad1	O
expression	O
on	O
thymocytes	B-cell_type
,	O
we	O
examined	O
thymic	O
selection	O
using	O
MHC	O
class	O
I-restricted	O
H-Y-TCR	O
transgenic	O
mice	O
.	O

Mad1	O
expression	O
in	O
vivo	O
reduces	O
the	O
efficiency	O
of	O
positive	O
selection	O
.	O

Furthermore	O
,	O
thymocytes	B-cell_type
and	O
splenic	B-cell_type
T	I-cell_type
cells	I-cell_type
from	O
lck-Mad1	O
transgenic	O
mice	O
display	O
a	O
profound	O
proliferative	O
defect	O
in	O
response	O
to	O
activation	O
with	O
either	O
PMA/Ionomycin	O
or	O
immobilized	B-protein
anti-CD3/CD28	I-protein
antibody	I-protein
.	O

This	O
proliferative	O
defect	O
is	O
not	O
reversed	O
by	O
addition	O
of	O
exogenous	B-protein
IL-2	I-protein
and	O
is	O
p53-independent	O
.	O

The	O
growth	O
inhibition	O
caused	O
by	O
Mad1	B-protein
is	O
overcome	O
by	O
expression	O
of	O
active	B-protein
c-Myc	I-protein
.	O

-DOCSTART-	O

Differential	O
requirement	O
for	O
the	O
transcription	B-protein
factor	I-protein
PU.1	I-protein
in	O
the	O
generation	O
of	O
natural	B-cell_type
killer	I-cell_type
cells	I-cell_type
versus	O
B	B-cell_type
and	I-cell_type
T	I-cell_type
cells	I-cell_type
.	O

PU.1	B-protein
is	O
a	O
member	O
of	O
the	O
Ets	B-protein
family	I-protein
of	I-protein
transcription	I-protein
factors	I-protein
required	O
for	O
the	O
development	O
of	O
various	O
lymphoid	B-cell_line
and	I-cell_line
myeloid	I-cell_line
cell	I-cell_line
lineages	I-cell_line
,	O
but	O
its	O
role	O
in	O
natural	B-cell_type
killer	I-cell_type
(	I-cell_type
NK	I-cell_type
)	I-cell_type
cell	I-cell_type
development	O
is	O
not	O
known	O
.	O

The	O
study	O
shows	O
that	O
PU.1	B-protein
is	O
expressed	O
in	O
NK	B-cell_type
cells	I-cell_type
and	O
that	O
,	O
on	O
cell	O
transfer	O
into	O
alymphoid	O
Rag2/gammac	O
(	O
-/-	O
)	O
mice	O
,	O
hematopoietic	B-cell_type
progenitors	I-cell_type
of	O
PU.1	B-cell_type
(	I-cell_type
-/-	I-cell_type
)	I-cell_type
fetal	I-cell_type
liver	I-cell_type
cells	I-cell_type
could	O
generate	O
functional	O
NK	B-cell_type
cells	I-cell_type
but	O
not	O
B	B-cell_type
or	I-cell_type
T	I-cell_type
cells	I-cell_type
.	O

Nevertheless	O
,	O
the	O
numbers	O
of	O
bone	B-cell_type
marrow	I-cell_type
NK	I-cell_type
cell	I-cell_type
precursors	I-cell_type
and	O
splenic	B-cell_type
mature	I-cell_type
NK	I-cell_type
cells	I-cell_type
were	O
reduced	O
compared	O
to	O
controls	O
.	O

Moreover	O
,	O
PU.1	B-cell_type
(	I-cell_type
-/-	I-cell_type
)	I-cell_type
NK	I-cell_type
cells	I-cell_type
displayed	O
reduced	O
expression	O
of	O
the	O
receptors	O
for	O
stem	B-protein
cell	I-protein
factor	I-protein
and	O
interleukin	B-protein
(	I-protein
IL	I-protein
)	I-protein
-7	I-protein
,	O
suggesting	O
a	O
nonredundant	O
role	O
for	O
PU.1	B-protein
in	O
regulating	O
the	O
expression	O
of	O
these	O
cytokine	B-DNA
receptor	I-DNA
genes	I-DNA
during	O
NK	O
cell	O
development	O
.	O

PU.1	B-cell_type
(	I-cell_type
-/-	I-cell_type
)	I-cell_type
NK	I-cell_type
cells	I-cell_type
also	O
showed	O
defective	O
expression	O
of	O
inhibitory	O
and	O
activating	O
members	O
of	O
the	O
Ly49	B-protein
family	I-protein
and	O
failed	O
to	O
proliferate	O
in	O
response	O
to	O
IL-2	B-protein
and	O
IL-12	B-protein
.	O

Thus	O
,	O
despite	O
the	O
less	O
stringent	O
requirement	O
for	O
PU.1	B-protein
in	O
NK	O
cell	O
development	O
compared	O
to	O
B	B-cell_type
and	I-cell_type
T	I-cell_type
cells	I-cell_type
,	O
PU.1	B-protein
regulates	O
NK	O
cell	O
differentiation	O
and	O
homeostasis	O
.	O

-DOCSTART-	O

Benzene-extracted	O
components	O
are	O
important	O
for	O
the	O
major	O
activity	O
of	O
diesel	O
exhaust	O
particles	O
:	O
effect	O
on	O
interleukin-8	B-DNA
gene	I-DNA
expression	O
in	O
human	B-cell_type
bronchial	I-cell_type
epithelial	I-cell_type
cells	I-cell_type
.	O

Epidemiologic	O
and	O
experimental	O
studies	O
suggest	O
that	O
diesel	O
exhaust	O
particles	O
(	O
DEPs	O
)	O
may	O
be	O
related	O
to	O
increasing	O
respiratory	O
mortality	O
and	O
morbidity	O
.	O

We	O
have	O
shown	O
that	O
DEPs	O
augmented	O
the	O
production	O
of	O
inflammatory	B-protein
cytokines	I-protein
by	O
human	B-cell_type
airway	I-cell_type
epithelial	I-cell_type
cells	I-cell_type
in	O
vitro	O
.	O

To	O
better	O
understand	O
the	O
mechanisms	O
of	O
their	O
proinflammatory	O
activities	O
,	O
we	O
studied	O
the	O
effects	O
of	O
several	O
components	O
extracted	O
from	O
DEPs	O
on	O
interleukin	O
(	O
IL	O
)	O
-8	O
expression	O
in	O
human	B-cell_line
bronchial	I-cell_line
epithelial	I-cell_line
cell	I-cell_line
line	I-cell_line
BEAS-2B	I-cell_line
and	O
normal	B-cell_type
human	I-cell_type
airway	I-cell_type
epithelial	I-cell_type
cells	I-cell_type
obtained	O
from	O
very	O
peripheral	O
airways	O
by	O
an	O
ultrathin	O
bronchoscope	O
.	O

We	O
used	O
several	O
agents	O
active	O
on	O
signal	O
transduction	O
pathways	O
in	O
cytokine	O
expression	O
,	O
such	O
as	O
the	O
protein	O
kinase	O
C	O
inhibitor	O
staurosporin	O
,	O
antioxidant	O
agents	O
including	O
N-acetyl	O
cysteine	O
(	O
NAC	O
)	O
and	O
pyrrolidine	O
dithiocarbamate	O
(	O
PDTC	O
)	O
,	O
and	O
p38	B-protein
mitogen-activated	I-protein
protein	I-protein
kinase	I-protein
(	O
MAPK	B-protein
)	O
inhibitor	O
SB203580	O
.	O

Benzene-extracted	O
components	O
showed	O
effects	O
mimicking	O
DEPs	O
on	O
IL-8	O
gene	O
expression	O
,	O
release	O
of	O
several	O
cytokines	B-protein
(	O
IL-8	B-protein
;	O
granulocyte	B-protein
macrophage	I-protein
colony-stimulating	I-protein
factor	I-protein
;	O
and	O
regulated	O
on	O
activation	O
,	O
normal	B-cell_type
T	I-cell_type
cells	I-cell_type
expressed	O
and	O
secreted	O
)	O
and	O
nuclear	O
factor	O
(	O
NF	O
)	O
-kappa	O
B	O
activation	O
.	O

We	O
also	O
found	O
that	O
NAC	O
,	O
PDTC	O
,	O
and	O
SB203580	O
suppressed	O
the	O
activities	O
of	O
DEPs	O
and	O
their	O
benzene	O
extracts	O
,	O
suggesting	O
the	O
roles	O
of	O
oxidants-mediated	O
NF-kappa	O
B	O
activation	O
and	O
p38MAPK	B-protein
pathways	O
.	O

Finally	O
,	O
benzo	O
[	O
a	O
]	O
pyrene	O
,	O
one	O
of	O
the	O
important	O
compounds	O
included	O
in	O
the	O
benzene	O
component	O
,	O
replicated	O
the	O
activities	O
shown	O
by	O
DEPs	O
.	O

-DOCSTART-	O

The	O
nuclear	B-protein
receptor	I-protein
PPAR	I-protein
gamma	I-protein
is	O
expressed	O
by	O
mouse	B-cell_type
T	I-cell_type
lymphocytes	I-cell_type
and	O
PPAR	O
gamma	O
agonists	O
induce	O
apoptosis	O
.	O

Peroxisome	B-protein
proliferator-activated	I-protein
receptor	I-protein
(	I-protein
PPAR	I-protein
)	I-protein
-gamma	I-protein
is	O
a	O
nuclear	B-protein
hormone	I-protein
receptor	I-protein
that	O
serves	O
as	O
a	O
trans	O
factor	O
to	O
regulate	O
lipid	O
metabolism	O
.	O

Intense	O
interest	O
is	O
focused	O
on	O
PPAR-gamma	B-protein
and	O
its	O
ligands	O
owing	O
to	O
its	O
putative	O
role	O
in	O
adipocyte	O
differentiation	O
.	O

Little	O
is	O
known	O
,	O
however	O
,	O
about	O
the	O
functions	O
of	O
PPAR-gamma	B-protein
in	O
the	O
immune	O
system	O
,	O
especially	O
in	O
T	B-cell_type
lymphocytes	I-cell_type
.	O

We	O
demonstrate	O
that	O
both	O
naive	B-cell_type
and	I-cell_type
activated	I-cell_type
ovalbumin-specific	I-cell_type
T	I-cell_type
cells	I-cell_type
from	O
DO11.10-transgenic	O
mice	O
express	O
PPAR-gamma	B-RNA
mRNA	O
and	O
protein	O
.	O

In	O
order	O
to	O
determine	O
the	O
function	O
of	O
PPAR-gamma	B-protein
,	O
T	B-cell_type
cells	I-cell_type
were	O
stimulated	O
with	O
phorbol	O
12-myristate	O
13-acetate	O
and	O
ionomycin	O
or	O
antigen	O
and	O
antigen-presenting	B-cell_type
cells	I-cell_type
.	O

Simultaneous	O
exposure	O
to	O
PPAR-gamma	O
ligands	O
(	O
e.g.	O
15-deoxy-Delta	O
(	O
12	O
,	O
14	O
)	O
-prostaglandin	O
J	O
(	O
2	O
)	O
,	O
troglitazone	O
)	O
showed	O
drastic	O
inhibition	O
of	O
proliferation	O
and	O
significant	O
decreases	O
in	O
cell	O
viability	O
.	O

The	O
decrease	O
in	O
cell	O
viability	O
was	O
due	O
to	O
apoptosis	O
of	O
the	O
T	B-cell_type
lymphocytes	I-cell_type
,	O
and	O
occurred	O
only	O
when	O
cells	O
were	O
treated	O
with	O
PPAR-gamma	B-protein
,	O
and	O
not	O
PPAR-alpha	O
agonists	O
,	O
revealing	O
specificity	O
of	O
this	O
response	O
for	O
PPAR-gamma	B-protein
.	O

These	O
observations	O
suggest	O
that	O
PPAR-gamma	O
agonists	O
play	O
an	O
important	O
role	O
in	O
regulating	O
T	O
cell-mediated	O
immune	O
responses	O
by	O
inducing	O
apoptosis	O
.	O

T	O
cell	O
death	O
via	O
PPAR-gamma	O
ligation	O
may	O
act	O
as	O
a	O
potent	O
anti-inflammatory	O
signal	O
in	O
the	O
immune	O
system	O
,	O
and	O
ligands	O
could	O
possibly	O
be	O
used	O
to	O
control	O
disorders	O
in	O
which	O
excessive	O
inflammation	O
occurs	O
.	O

-DOCSTART-	O

Suppression	O
of	O
lung	O
inflammation	O
in	O
rats	O
by	O
prevention	O
of	O
NF-kappaB	O
activation	O
in	O
the	O
liver	O
.	O

Activation	O
of	O
NF-kappaB	B-protein
and	O
production	O
of	O
NF-kappaB	B-protein
-dependent	O
chemokines	B-protein
are	O
thought	O
to	O
be	O
involved	O
in	O
the	O
pathogenesis	O
of	O
neutrophilic	O
lung	O
inflammation	O
.	O

Calpain-1	O
inhibitor	O
(	O
CI-1	O
)	O
blocks	O
activation	O
of	O
NF-kappaB	B-protein
by	O
preventing	O
proteolysis	O
of	O
the	O
inhibitory	O
protein	O
IkappaB-alpha	B-protein
by	O
the	O
ubiquitin/proteasome	O
pathway	O
.	O

We	O
hypothesized	O
that	O
inhibition	O
of	O
proteasome	O
function	O
with	O
CI-1	O
would	O
block	O
NF-kappaB	O
activation	O
in	O
vivo	O
after	O
intraperitoneal	O
(	O
i.p.	O
)	O
treatment	O
with	O
bacterial	O
lipopolysaccharide	O
(	O
LPS	O
)	O
,	O
and	O
that	O
NF-kappaB	O
inhibition	O
would	O
be	O
associated	O
with	O
suppression	O
of	O
chemokine	O
gene	O
expression	O
and	O
attenuation	O
of	O
neutrophilic	O
alveolitis	O
.	O

We	O
treated	O
rats	O
with	O
a	O
single	O
i.p.	O
injection	O
of	O
CI-1	O
(	O
10	O
mg/kg	O
)	O
two	O
hours	O
prior	O
to	O
i.p.	O
LPS	O
(	O
7	O
mg/kg	O
)	O
.	O

Treatment	O
with	O
Cl-1	O
prevented	O
degradation	O
of	O
IkappaB-alpha	B-protein
and	O
activation	O
of	O
NF-kappaB	B-protein
in	O
the	O
liver	O
in	O
response	O
to	O
LPS	O
;	O
however	O
,	O
Cl-1	O
treatment	O
had	O
no	O
detected	O
effect	O
on	O
NF-kappaB	O
activation	O
in	O
lung	O
tissue	O
.	O

CI-1	O
treatment	O
prior	O
to	O
LPS	O
resulted	O
in	O
40	O
%	O
lower	O
MIP-2	O
concentration	O
in	O
lung	O
lavage	O
fluid	O
compared	O
to	O
rats	O
treated	O
with	O
vehicle	O
prior	O
to	O
LPS	O
(	O
502	O
+/-	O
112	O
pg/ml	O
vs.	O
859	O
+/-144	O
pg/ml	O
,	O
P	O
<	O
0.05	O
)	O
.	O

In	O
addition	O
,	O
CI-1	O
treatment	O
substantially	O
inhibited	O
LPS-induced	O
neutrophilic	O
alveolitis	O
(	O
2.7+	O
/-	O
1.2	O
x	O
10	O
(	O
5	O
)	O
vs.	O
43.7	O
+/-	O
12.2	O
x	O
10	O
(	O
5	O
)	O
lung	B-cell_type
lavage	I-cell_type
neutrophils	I-cell_type
,	O
P	O
<	O
0.01	O
)	O
.	O

These	O
data	O
indicate	O
that	O
NF-kappaB	O
inhibition	O
in	O
the	O
liver	O
can	O
alter	O
lung	O
inflammation	O
induced	O
by	O
systemic	O
LPS	O
treatment	O
and	O
suggest	O
that	O
a	O
liver-lung	O
interaction	O
contributes	O
to	O
the	O
inflammatory	O
response	O
of	O
the	O
lung	O
.	O

-DOCSTART-	O

Notch1	B-protein
regulates	O
maturation	O
of	O
CD4+	B-cell_type
and	I-cell_type
CD8+	I-cell_type
thymocytes	I-cell_type
by	O
modulating	O
TCR	O
signal	O
strength	O
.	O

Notch	O
signaling	O
regulates	O
cell	O
fate	O
decisions	O
in	O
multiple	O
lineages	O
.	O

We	O
demonstrate	O
in	O
this	O
report	O
that	O
retroviral	O
expression	O
of	O
activated	O
Notch1	B-protein
in	O
mouse	B-cell_type
thymocytes	I-cell_type
abrogates	O
differentiation	O
of	O
immature	B-cell_type
CD4+CD8+	I-cell_type
thymocytes	I-cell_type
into	O
both	O
CD4	B-cell_type
and	I-cell_type
CD8	I-cell_type
mature	I-cell_type
single-positive	I-cell_type
T	I-cell_type
cells	I-cell_type
.	O

The	O
ability	O
of	O
Notch1	B-protein
to	O
inhibit	O
T	O
cell	O
development	O
was	O
observed	O
in	O
vitro	O
and	O
in	O
vivo	O
with	O
both	O
normal	B-cell_type
and	I-cell_type
TCR	I-cell_type
transgenic	I-cell_type
thymocytes	I-cell_type
.	O

Notch1	B-protein
-mediated	O
developmental	O
arrest	O
was	O
dose	O
dependent	O
and	O
was	O
associated	O
with	O
impaired	O
thymocyte	O
responses	O
to	O
TCR	O
stimulation	O
.	O

Notch1	B-protein
also	O
inhibited	O
TCR-mediated	O
signaling	O
in	O
Jurkat	B-cell_line
T	I-cell_line
cells	I-cell_line
.	O

These	O
data	O
indicate	O
that	O
constitutively	O
active	O
Notch1	B-protein
abrogates	O
CD4+	O
and	O
CD8+	O
maturation	O
by	O
interfering	O
with	O
TCR	O
signal	O
strength	O
and	O
provide	O
an	O
explanation	O
for	O
the	O
physiological	O
regulation	O
of	O
Notch	O
expression	O
during	O
thymocyte	O
development	O
.	O

-DOCSTART-	O

Expression	O
of	O
SART3	B-protein
antigen	I-protein
and	O
induction	O
of	O
CTLs	B-cell_type
by	O
SART3-derived	O
peptides	O
in	O
breast	O
cancer	O
patients	O
.	O

We	O
recently	O
reported	O
the	O
SART3	B-protein
tumour-rejection	I-protein
antigen	I-protein
as	O
possessing	O
tumour	O
epitopes	O
capable	O
of	O
inducing	O
HLA-class	B-cell_type
I-restricted	I-cell_type
cytotoxic	I-cell_type
T	I-cell_type
lymphocytes	I-cell_type
(	O
CTLs	B-cell_type
)	O
.	O

This	O
study	O
investigated	O
expression	O
of	O
the	O
SART3	B-protein
antigen	I-protein
in	O
breast	O
cancer	O
to	O
explore	O
an	O
appropriate	O
molecule	O
for	O
use	O
in	O
specific	O
immunotherapy	O
of	O
breast	O
cancer	O
patients	O
.	O

The	O
SART3	B-protein
antigen	I-protein
was	O
detected	O
in	O
all	O
of	O
the	O
breast	B-cell_line
cancer	I-cell_line
cell	I-cell_line
lines	I-cell_line
tested	O
,	O
30	O
of	O
40	O
(	O
75	O
%	O
)	O
breast	O
cancer	O
tissue	O
samples	O
,	O
and	O
0	O
of	O
3	O
non-tumourous	O
breast	O
tissue	O
samples	O
.	O

SART3	O
derived	O
peptides	O
at	O
positions	B-DNA
109-118	I-DNA
and	I-DNA
315-323	I-DNA
induced	O
HLA-A24	O
restricted	O
CTLs	B-cell_type
that	O
reacted	O
to	O
breast	B-cell_type
cancer	I-cell_type
cells	I-cell_type
from	O
the	O
peripheral	B-cell_type
blood	I-cell_type
mononuclear	I-cell_type
cells	I-cell_type
(	O
PBMCs	B-cell_type
)	O
of	O
breast	O
cancer	O
patients	O
.	O

Therefore	O
,	O
the	O
SART3	B-protein
antigen	I-protein
and	O
its	O
peptides	O
could	O
be	O
an	O
appropriate	O
molecule	O
for	O
use	O
in	O
specific	O
immunotherapy	O
of	O
the	O
majority	O
of	O
HLA-A24-positive	O
breast	O
cancer	O
patients	O

-DOCSTART-	O

Autostimulation	O
of	O
the	O
Epstein-Barr	O
virus	O
BRLF1	B-DNA
promoter	I-DNA
is	O
mediated	O
through	O
consensus	O
Sp1	B-DNA
and	I-DNA
Sp3	I-DNA
binding	I-DNA
sites	I-DNA
.	O

As	O
an	O
essential	O
step	O
in	O
the	O
lytic	O
cascade	O
,	O
the	O
Rta	O
homologues	O
of	O
gammaherpesviruses	O
all	O
activate	O
their	O
own	O
expression	O
.	O

Consistent	O
with	O
this	O
biologic	O
function	O
,	O
the	O
Epstein-Barr	O
virus	O
(	O
EBV	O
)	O
Rta	B-protein
protein	I-protein
powerfully	O
stimulates	O
the	O
promoter	B-DNA
of	O
its	O
own	O
gene	O
,	O
Rp	B-DNA
,	O
in	O
EBV-positive	B-cell_type
B	I-cell_type
cells	I-cell_type
in	O
transient-transfection	O
reporter-based	O
assays	O
.	O

We	O
analyzed	O
the	O
activity	O
of	O
RpCAT	B-DNA
in	O
response	O
to	O
Rta	B-protein
by	O
deletional	O
and	O
site-directed	O
mutagenesis	O
.	O

Two	O
cognate	B-DNA
Sp1	I-DNA
binding	I-DNA
sites	I-DNA
located	O
at	O
-279	O
and	O
-45	O
relative	O
to	O
the	O
transcriptional	B-DNA
start	I-DNA
site	I-DNA
proved	O
crucial	O
for	O
Rta-mediated	O
activation	O
.	O

Previously	O
described	O
binding	O
sites	O
for	O
the	O
cellular	O
transcription	B-protein
factor	I-protein
Zif268	I-protein
and	O
the	O
viral	B-protein
transactivator	I-protein
ZEBRA	I-protein
were	O
found	O
to	O
be	O
dispensable	O
for	O
activation	O
of	O
RpCAT	B-DNA
by	O
Rta	B-protein
.	O

Gel	O
shift	O
analysis	O
,	O
using	O
extracts	O
of	O
B	B-cell_type
cells	I-cell_type
in	O
latency	O
or	O
induced	O
into	O
the	O
lytic	O
cycle	O
,	O
identified	O
Sp1	B-protein
and	O
Sp3	B-protein
as	O
the	O
predominant	O
cellular	B-protein
proteins	I-protein
bound	O
to	O
Rp	B-DNA
near	O
-45	O
.	O

During	O
the	O
lytic	O
cycle	O
,	O
ZEBRA	B-protein
bound	O
Rp	B-DNA
near	O
the	O
Sp1/Sp3	B-DNA
site	I-DNA
.	O

The	O
binding	O
of	O
Sp1	B-protein
and	O
Sp3	B-protein
to	O
Rp	B-DNA
correlated	O
with	O
the	O
reporter	O
activities	O
in	O
the	O
mutagenesis	O
study	O
,	O
establishing	O
a	O
direct	O
link	O
between	O
transcriptional	O
activation	O
of	O
Rp	B-DNA
by	O
Rta	B-protein
and	O
DNA	O
binding	O
by	O
Sp1	B-protein
and/or	O
Sp3	B-protein
.	O

The	O
relative	O
abundance	O
or	O
functional	O
state	O
of	O
the	O
cellular	O
Sp1	B-protein
and	O
Sp3	B-protein
transcription	B-protein
factors	I-protein
may	O
be	O
altered	O
in	O
response	O
to	O
stimuli	O
that	O
induce	O
the	O
BRLF1	B-DNA
promoter	I-DNA
and	O
thereby	O
contribute	O
to	O
the	O
activation	O
of	O
the	O
viral	O
lytic	O
cycle	O
.	O

-DOCSTART-	O

Gene	O
transfer	O
of	O
antisense	O
hypoxia	B-protein
inducible	I-protein
factor-1	I-protein
alpha	I-protein
enhances	O
the	O
therapeutic	O
efficacy	O
of	O
cancer	O
immunotherapy	O
.	O

Solid	O
tumors	O
meet	O
their	O
demands	O
for	O
nascent	O
blood	O
vessels	O
and	O
increased	O
glycolysis	O
,	O
to	O
combat	O
hypoxia	O
,	O
by	O
activating	O
multiple	O
genes	O
involved	O
in	O
angiogenesis	O
and	O
glucose	O
metabolism	O
.	O

Hypoxia	B-protein
inducible	I-protein
factor-1	I-protein
(	O
HIF-1	B-protein
)	O
is	O
a	O
constitutively	O
expressed	O
basic	O
helix-loop-helix	B-protein
transcription	I-protein
factor	I-protein
,	O
formed	O
by	O
the	O
assembly	O
of	O
HIF-1alpha	B-protein
and	O
HIF-1beta	B-protein
(	O
Arnt	B-protein
)	O
,	O
that	O
is	O
stablized	O
in	O
response	O
to	O
hypoxia	O
,	O
and	O
rapidly	O
degraded	O
under	O
normoxic	O
conditions	O
.	O

It	O
activates	O
the	O
transcription	O
of	O
genes	O
important	O
for	O
maintaining	O
oxygen	O
homeostasis	O
.	O

Here	O
,	O
we	O
demonstrate	O
that	O
engineered	O
down-regulation	O
of	O
HIF-1alpha	B-protein
by	O
intratumoral	O
gene	O
transfer	O
of	O
an	O
antisense	B-DNA
HIF-1alpha	I-DNA
plasmid	I-DNA
leads	O
to	O
the	O
down-regulation	O
of	O
VEGF	B-DNA
,	O
and	O
decreased	O
tumor	O
microvessel	O
density	O
.	O

Antisense	O
HIF-1alpha	O
monotherapy	O
resulted	O
in	O
the	O
complete	O
and	O
permanent	O
rejection	O
of	O
small	O
(	O
0.1	O
cm	O
in	O
diameter	O
)	O
EL-4	O
tumors	O
,	O
which	O
is	O
unusual	O
for	O
an	O
anti-angiogenic	O
agent	O
where	O
transient	O
suppression	O
of	O
tumor	O
growth	O
is	O
the	O
norm	O
.	O

It	O
induced	O
NK	B-cell_type
cell	I-cell_type
-dependent	O
rejection	O
of	O
tumors	O
,	O
but	O
failed	O
to	O
stimulate	O
systemic	O
T	B-cell_type
cell	I-cell_type
-mediated	O
anti-tumor	O
immunity	O
,	O
and	O
synergized	O
with	O
B7-1-mediated	O
immunotherapy	O
to	O
cause	O
the	O
NK	B-cell_type
cell	I-cell_type
and	O
CD8	B-cell_type
T	I-cell_type
cell	I-cell_type
-dependent	O
rejection	O
of	O
larger	O
EL-4	O
tumors	O
(	O
0.4	O
cm	O
in	O
diameter	O
)	O
that	O
were	O
refractory	O
to	O
monotherapies	O
.	O

Mice	O
cured	O
of	O
their	O
tumors	O
by	O
combination	O
therapy	O
resisted	O
a	O
rechallenge	O
with	O
parental	B-cell_type
tumor	I-cell_type
cells	I-cell_type
,	O
indicating	O
systemic	O
antitumor	O
immunity	O
had	O
been	O
achieved	O
.	O

In	O
summary	O
,	O
whilst	O
intensive	O
investigations	O
are	O
in	O
progress	O
to	O
target	O
the	O
many	O
HIF-1	B-protein
effectors	O
,	O
the	O
results	O
herein	O
indicate	O
that	O
blocking	O
hypoxia-inducible	O
pathways	O
and	O
enhancing	O
NK-mediated	O
antitumor	O
immunity	O
by	O
targeting	O
HIF-1	B-protein
itself	O
may	O
be	O
advantageous	O
,	O
especially	O
when	O
combined	O
with	O
cancer	O
immunotherapy	O
.	O

-DOCSTART-	O

Runx2	B-DNA
:	O
a	O
novel	O
oncogenic	O
effector	O
revealed	O
by	O
in	O
vivo	O
complementation	O
and	O
retroviral	O
tagging	O
.	O

The	O
Runx2	B-DNA
(	O
Cbfa1	B-DNA
,	O
Pebp2alphaA	B-DNA
,	O
Aml3	B-DNA
)	O
gene	O
was	O
previously	O
identified	O
as	O
a	O
frequent	O
target	O
for	O
transcriptional	O
activation	O
by	O
proviral	O
insertion	O
in	O
T-cell	O
lymphomas	O
of	O
CD2-MYC	O
transgenic	O
mice	O
.	O

We	O
have	O
recently	O
shown	O
that	O
over-expression	O
of	O
the	O
full-length	O
,	O
most	O
highly	O
expressed	O
Runx2	B-DNA
isoform	O
in	O
the	O
thymus	O
perturbs	O
T-cell	O
development	O
,	O
leads	O
to	O
development	O
of	O
spontaneous	O
lymphomas	O
at	O
low	O
frequency	O
and	O
is	O
strongly	O
synergistic	O
with	O
Myc	B-protein
.	O

To	O
gain	O
further	O
insight	O
into	O
the	O
relationship	O
of	O
Runx2	B-DNA
to	O
other	O
lymphomagenic	O
pathways	O
,	O
we	O
tested	O
the	O
effect	O
of	O
combining	O
the	O
CD2-Runx2	B-DNA
transgene	I-DNA
either	O
with	O
a	O
Pim1	B-DNA
transgene	I-DNA
(	O
E	B-DNA
(	I-DNA
mu	I-DNA
)	I-DNA
-Pim1	I-DNA
)	O
or	O
with	O
the	O
p53	B-DNA
null	O
genotype	O
,	O
as	O
each	O
of	O
these	O
displays	O
independent	O
synergy	O
with	O
Myc	B-protein
.	O

In	O
both	O
cases	O
we	O
observed	O
synergistic	O
tumour	O
development	O
.	O

However	O
,	O
Runx2	B-protein
appeared	O
to	O
have	O
a	O
dominant	O
effect	O
on	O
the	O
tumour	O
phenotype	O
in	O
each	O
case	O
,	O
with	O
most	O
tumours	O
conforming	O
to	O
the	O
CD3	O
(	O
+	O
)	O
,	O
CD8	O
(	O
+	O
)	O
,	O
CD4	O
(	O
+/-	O
)	O
phenotype	O
seen	O
in	O
CD2-Runx2	O
mice	O
.	O

Neonatal	O
infection	O
of	O
CD2-Runx2	O
mice	O
with	O
Moloney	O
murine	O
leukaemia	O
virus	O
(	O
Moloney	O
MLV	O
)	O
also	O
led	O
to	O
a	O
dramatic	O
acceleration	O
of	O
tumour	O
onset	O
.	O

Analysis	O
of	O
known	O
Moloney	B-DNA
MLV	I-DNA
target	I-DNA
genes	I-DNA
in	O
these	O
lymphomas	O
showed	O
a	O
high	O
frequency	O
of	O
rearrangement	O
at	O
c-Myc	B-DNA
or	O
N-Myc	B-DNA
(	O
82	O
%	O
)	O
,	O
and	O
a	O
significant	O
number	O
at	O
Pim1	B-DNA
or	O
Pim2	B-DNA
(	O
23	O
%	O
)	O
,	O
and	O
at	O
Pal1/Gfi1	B-DNA
(	O
18	O
%	O
)	O
.	O

These	O
results	O
indicate	O
that	O
Runx2	B-protein
makes	O
a	O
distinct	O
contribution	O
to	O
T-cell	O
lymphoma	O
development	O
which	O
does	O
not	O
coincide	O
with	O
any	O
of	O
the	O
oncogene	O
complementation	O
groups	O
previously	O
identified	O
by	O
retroviral	O
tagging	O
.	O

-DOCSTART-	O

The	O
involvement	O
of	O
TNF-alpha	B-protein
-related	O
apoptosis-inducing	O
ligand	O
in	O
the	O
enhanced	O
cytotoxicity	O
of	O
IFN-beta	B-protein
-stimulated	O
human	B-cell_type
dendritic	I-cell_type
cells	I-cell_type
to	O
tumor	B-cell_type
cells	I-cell_type
.	O

TNF-alpha	B-protein
-related	O
apoptosis-inducing	O
ligand	O
(	O
TRAIL	O
)	O
is	O
characterized	O
by	O
its	O
preferential	O
induction	O
of	O
apoptosis	O
of	O
tumor	B-cell_type
cells	I-cell_type
but	O
not	O
normal	B-cell_type
cells	I-cell_type
.	O

Dendritic	B-cell_type
cells	I-cell_type
(	O
DCs	B-cell_type
)	O
,	O
besides	O
their	O
role	O
as	O
APCs	O
,	O
now	O
have	O
been	O
demonstrated	O
to	O
exert	O
cytotoxicity	O
or	O
cytostasis	O
on	O
some	O
tumor	B-cell_type
cells	I-cell_type
.	O

Here	O
,	O
we	O
report	O
that	O
both	O
human	B-cell_type
CD34	I-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
stem	I-cell_type
cell	I-cell_type
-derived	O
DCs	B-cell_type
(	O
CD34DCs	B-cell_type
)	O
and	O
human	B-cell_type
CD14	I-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
monocyte	I-cell_type
-derived	O
DCs	B-cell_type
(	O
MoDCs	B-cell_type
)	O
express	O
TRAIL	O
and	O
exhibit	O
cytotoxicity	O
to	O
some	O
types	O
of	O
tumor	B-cell_type
cells	I-cell_type
partially	O
through	O
TRAIL	O
.	O

Moderate	O
expression	O
of	O
TRAIL	O
appeared	O
on	O
CD34DCs	B-cell_type
from	O
the	O
8th	O
day	O
of	O
culture	O
and	O
was	O
also	O
seen	O
on	O
freshly	B-cell_type
isolated	I-cell_type
monocytes	I-cell_type
.	O

The	O
level	O
of	O
TRAIL	O
expression	O
remained	O
constant	O
until	O
DC	O
maturation	O
.	O

TRAIL	O
expression	O
on	O
immature	B-cell_type
CD34DCs	I-cell_type
or	O
MoDCs	B-cell_type
was	O
greatly	O
up-regulated	O
after	O
IFN-beta	B-protein
stimulation	O
.	O

Moreover	O
,	O
IFN-beta	B-protein
could	O
strikingly	O
enhance	O
the	O
ability	O
of	O
CD34DCs	B-cell_type
or	O
MoDCs	B-cell_type
to	O
kill	O
TRAIL-sensitive	O
tumor	B-cell_type
cells	I-cell_type
,	O
but	O
LPS	O
did	O
not	O
have	O
such	O
an	O
effect	O
.	O

The	O
up-regulation	O
of	O
TRAIL	O
on	O
IFN-beta	B-protein
-stimulated	O
DCs	B-cell_type
partially	O
contributed	O
to	O
the	O
increased	O
cytotoxicity	O
of	O
DCS	B-cell_type
:	O
Pretreatment	O
of	O
TRAIL-sensitive	O
tumor	B-cell_type
cells	I-cell_type
with	O
caspase-3	O
inhibitor	O
could	O
significantly	O
increase	O
their	O
resistance	O
to	O
the	O
cytotoxicity	O
of	O
IFN-beta	B-protein
-stimulated	O
DCS	B-cell_type
:	O
In	O
contrast	O
,	O
NF-kappaB	O
inhibitor	O
could	O
significantly	O
increase	O
the	O
sensitivity	O
of	O
tumor	B-cell_type
cells	I-cell_type
to	O
the	O
killing	O
by	O
nonstimulated	O
or	O
LPS-stimulated	O
DCS	B-cell_type
:	O
Our	O
studies	O
demonstrate	O
that	O
IFN-beta	B-protein
-stimulated	O
DCs	B-cell_type
are	O
functionally	O
cytotoxic	O
.	O

Thus	O
,	O
an	O
innate	O
mechanism	O
of	O
DC-mediated	O
antitumor	O
immunity	O
might	O
exist	O
in	O
vivo	O
in	O
which	O
DCs	B-cell_type
act	O
as	O
effectors	O
to	O
directly	O
kill	O
tumor	B-cell_type
cells	I-cell_type
partially	O
via	O
TRAIL	O
.	O

Subsequently	O
,	O
DCs	B-cell_type
act	O
as	O
APCs	O
involved	O
in	O
the	O
uptake	O
,	O
processing	O
,	O
and	O
presentation	O
of	O
apoptotic	B-protein
tumor	I-protein
Ags	I-protein
to	O
cross-prime	O
CD8	B-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
CTL	I-cell_type
cells	I-cell_type
.	O

-DOCSTART-	O

Tumor	B-cell_type
cells	I-cell_type
regulate	O
the	O
lytic	O
activity	O
of	O
tumor-specific	B-cell_type
cytotoxic	I-cell_type
t	I-cell_type
lymphocytes	I-cell_type
by	O
modulating	O
the	O
inhibitory	O
natural	O
killer	O
receptor	O
function	O
.	O

Tumor-infiltrating	B-cell_type
p58+	I-cell_type
T	I-cell_type
cells	I-cell_type
from	O
a	O
renal	O
tumor	O
were	O
specifically	O
expanded	O
in	O
response	O
to	O
tumor	O
cell	O
stimulation	O
and	O
cloned	O
.	O

These	O
p58+	B-cell_type
T	I-cell_type
cells	I-cell_type
were	O
found	O
to	O
express	O
a	O
memory	B-cell_type
phenotype	I-cell_type
and	O
corresponded	O
to	O
clonal	O
TCRBV3	O
T-cell	O
expansion	O
.	O

Functionally	O
,	O
p58	B-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
CTLs	I-cell_type
displayed	O
a	O
low	O
lytic	O
activity	O
for	O
HLA-A2	B-cell_type
tumor	I-cell_type
and	I-cell_type
normal	I-cell_type
cells	I-cell_type
.	O

However	O
,	O
this	O
lytic	O
activity	O
was	O
significantly	O
increased	O
after	O
blockade	O
of	O
p58	B-protein
with	O
specific	O
monoclonal	B-protein
antibodies	I-protein
.	O

Interestingly	O
,	O
we	O
demonstrated	O
that	O
stimulation	O
by	O
tumor	B-cell_type
cells	I-cell_type
was	O
required	O
to	O
trigger	O
the	O
inhibitory	O
effect	O
of	O
p58	B-protein
on	O
the	O
lytic	O
activity	O
of	O
antigen-specific	B-cell_type
CTLs	I-cell_type
and	O
that	O
stimulation	O
of	O
the	O
inhibitory	O
function	O
of	O
p58	B-protein
by	O
tumor	B-cell_type
cells	I-cell_type
correlated	O
with	O
an	O
inhibition	O
of	O
nuclear	O
factor-kappaB	O
activation	O
in	O
p58+	B-cell_type
tumor-specific	I-cell_type
CTLS	I-cell_type
.	O

-DOCSTART-	O

T-cell	B-protein
factor-1	I-protein
expression	O
during	O
human	B-cell_type
natural	I-cell_type
killer	I-cell_type
cell	I-cell_type
development	O
and	O
in	O
circulating	O
CD56	B-protein
(	I-protein
+	I-protein
)	I-protein
bright	O
natural	B-cell_type
killer	I-cell_type
cells	I-cell_type
.	O

Transcription	B-protein
factors	I-protein
are	O
essential	O
to	O
govern	O
differentiation	O
along	O
the	O
lymphoid	B-cell_type
lineage	I-cell_type
from	O
uncommitted	B-cell_type
hematopoietic	I-cell_type
stem	I-cell_type
cells	I-cell_type
.	O

Although	O
many	O
of	O
these	O
transcription	B-protein
factors	I-protein
have	O
putative	O
roles	O
based	O
on	O
murine	O
knockout	O
experiments	O
,	O
their	O
function	O
in	O
human	O
lymphoid	O
development	O
is	O
less	O
known	O
and	O
was	O
studied	O
further	O
.	O

Transcription	O
factor	O
expression	O
in	O
fresh	O
and	O
cultured	O
adult	B-cell_type
human	I-cell_type
bone	I-cell_type
marrow	I-cell_type
and	I-cell_type
umbilical	I-cell_type
cord	I-cell_type
blood	I-cell_type
progenitors	I-cell_type
was	O
evaluated	O
.	O

We	O
found	O
that	O
fresh	B-cell_type
CD34	I-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
Lin	I-cell_type
(	I-cell_type
-	I-cell_type
)	I-cell_type
cells	I-cell_type
that	O
are	O
human	O
leukocyte	O
antigen	O
(	O
HLA	O
)	O
-DR	O
(	O
-	O
)	O
or	O
CD38	O
(	O
-	O
)	O
constitutively	O
express	O
GATA-3	B-protein
but	O
not	O
T-cell	B-protein
factor-1	I-protein
(	O
TCF-1	B-protein
)	O
or	O
Id-3	B-protein
.	O

Culture	O
with	O
the	O
murine	B-cell_line
fetal	I-cell_line
liver	I-cell_line
cell	I-cell_line
line	I-cell_line
AFT024	I-cell_line
and	O
defined	O
cytokines	B-protein
was	O
capable	O
of	O
inducing	O
TCF-1	B-RNA
mRNA	I-RNA
.	O

However	O
,	O
no	O
T-cell	B-DNA
receptor	I-DNA
gene	I-DNA
rearrangement	O
was	O
identified	O
in	O
cultured	O
progeny	O
.	O

Id-3	B-protein
,	O
a	O
basic	B-protein
helix	I-protein
loop	I-protein
helix	I-protein
factor	I-protein
with	O
dominant	O
negative	O
function	O
for	O
T-cell	O
differentiation	O
transcription	B-protein
factors	I-protein
,	O
also	O
was	O
upregulated	O
and	O
may	O
explain	O
unsuccessful	O
T-cell	O
maturation	O
.	O

To	O
better	O
understand	O
the	O
developmental	O
link	O
between	O
natural	B-cell_type
killer	I-cell_type
(	I-cell_type
NK	I-cell_type
)	I-cell_type
cells	I-cell_type
derived	O
from	O
progenitors	B-cell_type
,	O
we	O
studied	O
NK	B-cell_type
cell	I-cell_type
subsets	I-cell_type
circulating	O
in	O
blood	O
.	O

CD56	B-cell_type
(	I-cell_type
+bright	I-cell_type
)	I-cell_type
,	O
but	O
not	O
CD56	B-cell_type
(	I-cell_type
+dim	I-cell_type
)	I-cell_type
,	O
NK	B-cell_type
cells	I-cell_type
constitutively	O
express	O
TCF-1	B-protein
by	O
reverse	O
transcriptase	O
polymerase	O
chain	O
reaction	O
and	O
Western	O
blot	O
analysis	O
.	O

The	O
TCF-1	B-protein
isoform	I-protein
found	O
in	O
CD56	B-cell_type
(	I-cell_type
+bright	I-cell_type
)	I-cell_type
cells	I-cell_type
,	O
which	O
express	O
lectin	B-protein
but	O
not	O
immunoglobulin	B-protein
class	I-protein
I	I-protein
recognizing	O
inhibitory	B-protein
receptors	I-protein
,	O
was	O
identical	O
to	O
that	O
induced	O
in	O
NK	O
cell	O
differentiation	O
culture	O
and	O
was	O
distinctly	O
different	O
from	O
isoforms	O
in	O
T	B-cell_type
cells	I-cell_type
.	O

These	O
results	O
suggest	O
that	O
TCF-1	B-protein
does	O
not	O
target	O
human	B-DNA
killer	I-DNA
immunoglobulin	I-DNA
receptor	I-DNA
genes	I-DNA
,	O
TCF-1	B-protein
is	O
uniquely	O
expressed	O
in	O
circulating	O
CD56	B-cell_type
(	I-cell_type
+bright	I-cell_type
)	I-cell_type
NK	I-cell_type
cells	I-cell_type
,	O
and	O
specific	O
TCF-1	B-protein
isoforms	I-protein
may	O
play	O
an	O
important	O
role	O
in	O
regulating	O
NK	O
differentiation	O
from	O
a	O
common	O
NK/T-cell	B-cell_type
progenitor	I-cell_type
.	O

-DOCSTART-	O

Transcriptional	O
activation	O
by	O
a	O
matrix	B-protein
associating	I-protein
region-binding	I-protein
protein	I-protein
.	O

contextual	O
requirements	O
for	O
the	O
function	O
of	O
bright	B-protein
.	O

Bright	B-protein
(	O
B	B-protein
cell	I-protein
regulator	I-protein
of	I-protein
IgH	I-protein
transcription	I-protein
)	O
is	O
a	O
B	B-cell_type
cell	I-cell_type
-specific	O
,	O
matrix	B-protein
associating	I-protein
region-binding	I-protein
protein	I-protein
that	O
transactivates	O
gene	O
expression	O
from	O
the	O
IgH	B-DNA
intronic	I-DNA
enhancer	I-DNA
(	O
E	B-DNA
mu	I-DNA
)	O
.	O

We	O
show	O
here	O
that	O
Bright	B-protein
has	O
multiple	O
contextual	O
requirements	O
to	O
function	O
as	O
a	O
transcriptional	B-protein
activator	I-protein
.	O

Bright	B-protein
can	O
not	O
transactivate	O
via	O
out	O
of	O
context	O
,	O
concatenated	B-protein
binding	I-protein
sites	I-protein
.	O

Transactivation	O
is	O
maximal	O
on	O
integrated	O
substrates	O
.	O

Two	O
of	O
the	O
three	O
previously	O
identified	O
binding	B-DNA
sites	I-DNA
in	O
E	B-DNA
mu	I-DNA
are	O
required	O
for	O
full	O
Bright	B-protein
transactivation	O
.	O

The	O
Bright	B-protein
DNA	I-protein
binding	I-protein
domain	I-protein
defined	O
a	O
new	O
family	O
,	O
which	O
includes	O
SWI1	B-protein
,	O
a	O
component	O
of	O
the	O
SWI.SNF	B-protein
complex	I-protein
shown	O
to	O
have	O
high	O
mobility	O
group-like	O
DNA	O
binding	O
characteristics	O
.	O

Similar	O
to	O
one	O
group	O
of	O
high	B-protein
mobility	I-protein
group	I-protein
box	I-protein
proteins	I-protein
,	O
Bright	B-protein
distorts	O
E	B-DNA
mu	I-DNA
binding	I-DNA
site	I-DNA
-containing	O
DNA	O
on	O
binding	O
,	O
supporting	O
the	O
concept	O
that	O
it	O
mediates	O
E	B-DNA
mu	I-DNA
remodeling	O
.	O

Transfection	O
studies	O
further	O
implicate	O
Bright	B-protein
in	O
facilitating	O
spatially	O
separated	O
promoter-enhancer	O
interactions	O
in	O
both	O
transient	O
and	O
stable	O
assays	O
.	O

Finally	O
,	O
we	O
show	O
that	O
overexpression	O
of	O
Bright	B-protein
leads	O
to	O
enhanced	O
DNase	O
I	O
sensitivity	O
of	O
the	O
endogenous	O
E	B-DNA
mu	I-DNA
matrix	O
associating	O
regions	O
.	O

These	O
data	O
further	O
suggest	O
that	O
Bright	B-protein
may	O
contribute	O
to	O
increased	O
gene	O
expression	O
by	O
remodeling	O
the	O
immunoglobulin	B-DNA
locus	I-DNA
during	O
B	B-cell_type
cell	I-cell_type
development	O
.	O

-DOCSTART-	O

Analysis	O
of	O
BCL-6	O
mutations	O
in	O
classic	O
Hodgkin	O
disease	O
of	O
the	O
B-	B-cell_type
and	I-cell_type
T-cell	I-cell_type
type	I-cell_type
.	O

BCL-6	B-DNA
is	O
essential	O
for	O
germinal	O
center	O
formation	O
and	O
thus	O
for	O
affinity	O
maturation	O
of	O
immunoglobulin	B-DNA
(	I-DNA
Ig	I-DNA
)	I-DNA
genes	I-DNA
by	O
somatic	O
mutations	O
.	O

The	O
5'-noncoding	B-DNA
region	I-DNA
of	O
the	O
BCL-6	B-DNA
gene	I-DNA
is	O
even	O
a	O
target	O
for	O
the	O
mutation	O
machinery	O
.	O

Translocations	O
of	O
the	O
BCL-6	B-DNA
gene	I-DNA
to	O
heterologous	B-DNA
promoters	I-DNA
and	O
mutations	O
of	O
its	O
5'-noncoding	B-DNA
regulatory	I-DNA
region	I-DNA
were	O
reported	O
to	O
be	O
potential	O
mechanisms	O
for	O
deregulating	O
BCL-6	B-DNA
expression	O
and	O
for	O
playing	O
a	O
role	O
in	O
the	O
genesis	O
of	O
non-Hodgkin	O
lymphoma	O
.	O

In	O
line	O
with	O
this	O
hypothesis	O
is	O
the	O
observation	O
that	O
B-cell	O
lymphoma	O
with	O
somatic	O
mutations	O
,	O
such	O
as	O
diffuse	O
large	O
B-cell	O
lymphoma	O
and	O
follicular	O
lymphoma	O
,	O
also	O
carry	O
BCL-6	O
mutations	O
,	O
some	O
of	O
which	O
are	O
recurrently	O
detectable	O
.	O

Classic	O
Hodgkin	O
disease	O
(	O
cHD	O
)	O
is	O
also	O
derived	O
from	O
B	B-cell_type
cells	I-cell_type
with	O
high	O
loads	O
of	O
somatic	O
mutations	O
and	O
thus	O
a	O
further	O
candidate	O
for	O
BCL-6	O
mutations	O
.	O

To	O
determine	O
the	O
presence	O
and	O
potential	O
role	O
of	O
BCL-6	O
mutations	O
in	O
cHD	O
,	O
the	O
5'-noncoding	B-DNA
BCL-6	I-DNA
proportion	O
of	O
single	B-cell_type
Hodgkin	I-cell_type
and	I-cell_type
Reed-Sternberg	I-cell_type
(	I-cell_type
HRS	I-cell_type
)	I-cell_type
cells	I-cell_type
from	O
6	O
cases	O
of	O
cHD	O
and	O
6	O
cases	O
of	O
HD-derived	B-cell_line
cell	I-cell_line
lines	I-cell_line
was	O
analyzed	O
.	O

All	O
B-cell-derived	O
HD	O
cases	O
and	O
cell	O
lines	O
harbored	O
BCL-6	O
mutations	O
.	O

In	O
contrast	O
,	O
both	O
T-cell-derived	O
HD	O
cases	O
and	O
cell	O
lines	O
were	O
devoid	O
of	O
BCL-6	O
mutations	O
.	O

With	O
only	O
one	O
exception	O
,	O
there	O
were	O
no	O
lymphoma-specific	O
recurrent	O
BCL-6	O
mutations	O
detected	O
,	O
and	O
BCL-6	B-protein
protein	I-protein
was	O
absent	O
from	O
the	O
HRS	B-cell_type
cells	I-cell_type
of	O
most	O
cases	O
.	O

In	O
conclusion	O
,	O
(	O
1	O
)	O
somatic	O
BCL-6	O
mutations	O
are	O
restricted	O
to	O
cHD	O
cases	O
of	O
B-cell	B-cell_type
origin	O
,	O
and	O
(	O
2	O
)	O
the	O
BCL-6	O
mutations	O
represent	O
mostly	O
irrelevant	O
somatic	O
base	O
substitutions	O
without	O
consequences	O
for	O
BCL-6	B-protein
protein	I-protein
expression	O
and	O
the	O
pathogenesis	O
of	O
cHD	O
.	O

-DOCSTART-	O

Gadd45gamma	B-DNA
is	O
dispensable	O
for	O
normal	O
mouse	O
development	O
and	O
T-cell	O
proliferation	O
.	O

Gadd45gamma	B-DNA
,	O
a	O
family	O
member	O
of	O
the	O
growth	B-DNA
arrest	I-DNA
and	I-DNA
DNA	I-DNA
damage-inducible	I-DNA
gene	I-DNA
family	I-DNA
45	I-DNA
(	O
Gadd45	B-DNA
)	O
,	O
is	O
strongly	O
induced	O
by	O
interleukin-2	B-protein
(	O
IL-2	B-protein
)	O
in	O
peripheral	B-cell_type
T	I-cell_type
cells	I-cell_type
.	O

While	O
in	O
most	O
tissues	O
all	O
Gadd45	B-DNA
family	I-DNA
members	I-DNA
are	O
expressed	O
,	O
Gadd45gamma	B-DNA
is	O
the	O
only	O
member	O
that	O
is	O
induced	O
by	O
IL-2	B-protein
.	O

Here	O
we	O
show	O
that	O
the	O
IL-2	B-protein
-induced	O
expression	O
of	O
Gadd45gamma	B-DNA
is	O
dependent	O
on	O
a	O
signaling	O
pathway	O
mediated	O
by	O
the	O
tyrosine	B-protein
kinase	I-protein
Jak3	I-protein
and	O
the	O
transcription	B-protein
factors	I-protein
Stat5a	B-protein
and	O
Stat5b	B-protein
(	O
signal	B-protein
transducer	I-protein
and	I-protein
activator	I-protein
of	I-protein
transcription	I-protein
)	O
.	O

Previous	O
studies	O
with	O
ectopically	O
overexpressed	O
Gadd45gamma	B-DNA
in	O
various	O
cell	O
lines	O
implicated	O
its	O
function	O
in	O
negative	O
growth	O
control	O
.	O

To	O
analyze	O
the	O
physiological	O
role	O
of	O
Gadd45gamma	B-DNA
we	O
used	O
homologous	O
recombination	O
to	O
generate	O
mice	O
lacking	O
Gadd45gamma	B-DNA
.	O

Gadd45gamma	B-DNA
-deficient	O
mice	O
develop	O
normally	O
,	O
are	O
indistinguishable	O
from	O
their	O
littermates	O
,	O
and	O
are	O
fertile	O
.	O

Furthermore	O
,	O
hematopoiesis	O
in	O
mice	O
lacking	O
Gadd45gamma	B-DNA
is	O
not	O
impaired	O
and	O
Gadd45gamma	B-DNA
-deficient	O
T	B-cell_type
lymphocytes	I-cell_type
show	O
normal	O
responses	O
to	O
IL-2	B-protein
.	O

These	O
data	O
demonstrate	O
that	O
Gadd45gamma	B-DNA
is	O
not	O
essential	O
for	O
normal	O
mouse	O
development	O
and	O
hematopoiesis	O
,	O
possibly	O
due	O
to	O
functional	O
redundancy	O
among	O
the	O
Gadd45	B-DNA
family	I-DNA
members	I-DNA
.	O

Gadd45gamma	B-DNA
is	O
also	O
dispensable	O
for	O
IL-2	B-protein
-induced	O
T-cell	O
proliferation	O
.	O

-DOCSTART-	O

Decreased	O
expression	O
of	O
c-myc	B-DNA
family	I-DNA
genes	I-DNA
in	O
thymuses	O
from	O
myasthenia	O
gravis	O
patients	O
.	O

The	O
thymus	O
is	O
a	O
critical	O
organ	O
for	O
the	O
elimination	O
of	O
autoreactive	B-cell_type
T	I-cell_type
cells	I-cell_type
by	O
apoptosis	O
.	O

We	O
studied	O
the	O
expression	O
of	O
apoptosis-associated	O
genes	O
,	O
bcl-xL	B-DNA
,	O
bad	B-DNA
,	O
caspase-3	B-DNA
,	O
and	O
c-myc	B-DNA
family	I-DNA
genes	I-DNA
in	O
myasthenia	O
gravis	O
(	O
MG	O
)	O
thymuses	O
.	O

We	O
observed	O
that	O
the	O
mRNA	O
levels	O
of	O
myc	B-DNA
family	I-DNA
genes	I-DNA
,	O
c-myc	B-DNA
and	O
max	B-DNA
,	O
were	O
markedly	O
reduced	O
in	O
MG	O
thymuses	O
.	O

These	O
results	O
indicate	O
that	O
c-myc	B-DNA
-mediated	O
signaling	O
is	O
abnormal	O
in	O
MG	O
thymuses	O
.	O

The	O
levels	O
of	O
molecules	O
whose	O
expressions	O
are	O
associated	O
with	O
myc	B-DNA
,	O
such	O
as	O
STAM	B-protein
,	O
prothymosin-alpha	B-protein
,	O
and	O
NFkappaB	B-protein

-DOCSTART-	O

Immunohistochemical	O
detection	O
of	O
interferon-gamma-producing	B-cell_type
cells	I-cell_type
in	O
dermatophytosis	O
.	O

Skin	O
lesions	O
of	O
dermatophytosis	O
are	O
thought	O
to	O
be	O
a	O
result	O
of	O
a	O
T	B-cell_type
cell	I-cell_type
-dependent	O
inflammatory	O
response	O
that	O
is	O
mediated	O
by	O
various	O
cytokines	B-protein
.	O

We	O
examined	O
whether	O
IFN-gamma-positive	B-cell_type
cells	I-cell_type
(	O
as	O
expression	O
of	O
Th1	B-cell_type
response	O
)	O
were	O
present	O
in	O
the	O
skin	O
lesions	O
of	O
dermatophytosis	O
in	O
situ	O
by	O
immunohistochemical	O
techniques	O
.	O

Mixtures	O
of	O
CD4-positive	B-cell_type
T	I-cell_type
cells	I-cell_type
and	O
CD8-positive	B-cell_type
T	I-cell_type
cells	I-cell_type
were	O
found	O
to	O
be	O
present	O
in	O
the	O
dermal	O
infiltrates	O
of	O
the	O
lesions	O
.	O

Considerable	O
numbers	O
of	O
CD1a-positive	B-cell_type
cells	I-cell_type
were	O
detected	O
in	O
the	O
upper	O
dermis	O
and	O
epidermis	O
.	O

A	O
marked	O
accumulation	O
of	O
CD68-positive	B-cell_type
cells	I-cell_type
was	O
found	O
in	O
the	O
upper	O
dermis	O
.	O

IFN-gamma-positive	B-cell_type
cells	I-cell_type
were	O
present	O
in	O
the	O
upper	O
dermis	O
of	O
the	O
lesions	O
.	O

The	O
pattern	O
of	O
IFN-gamma	B-protein
staining	O
appeared	O
to	O
be	O
intracellular	O
in	O
mononuclear	B-cell_type
lymphoid	I-cell_type
cells	I-cell_type
.	O

The	O
staining	O
was	O
considered	O
to	O
be	O
highly	O
specific	O
because	O
it	O
could	O
be	O
completely	O
blocked	O
by	O
preabsorption	O
with	O
recombinant	B-protein
IFN-gamma	I-protein
.	O

Our	O
data	O
support	O
the	O
hypothesis	O
that	O
the	O
skin	O
lesions	O
of	O
dermatophytosis	O
may	O
be	O
associated	O
with	O
a	O
Th1	B-cell_type
response	O
.	O

Th1	B-cell_type
response	O
,	O
which	O
is	O
characterized	O
by	O
IFN-gamma	B-protein
release	O
,	O
is	O
thought	O
to	O
be	O
involved	O
in	O
the	O
host	O
defense	O
against	O
dermatophytes	O
and	O
to	O
reflect	O
cutaneous	O
reaction	O
in	O
dermatophytosis	O
.	O

-DOCSTART-	O

Regulation	O
of	O
the	O
helix-loop-helix	B-protein
proteins	I-protein
,	O
E2A	B-protein
and	O
Id3	B-protein
,	O
by	O
the	O
Ras	B-protein
-	O
ERK	B-protein
MAPK	B-protein
cascade	O
.	O

Activation	O
of	O
mitogen-activated	B-protein
protein	I-protein
kinase	I-protein
(	O
MAPK	B-protein
)	O
pathways	O
leads	O
to	O
cellular	O
differentiation	O
and/or	O
proliferation	O
in	O
a	O
wide	O
variety	O
of	O
cell	O
types	O
,	O
including	O
developing	B-cell_type
thymocytes	I-cell_type
.	O

The	O
basic	B-protein
helix-loop-helix	I-protein
(	I-protein
bHLH	I-protein
)	I-protein
proteins	I-protein
E12	B-protein
and	O
E47	B-protein
and	O
an	O
inhibitor	B-protein
HLH	I-protein
protein	I-protein
,	O
Id3	B-protein
,	O
play	O
key	O
roles	O
in	O
thymocyte	O
differentiation	O
.	O

We	O
show	O
here	O
that	O
E2A	B-protein
DNA	O
binding	O
is	O
lowered	O
in	O
primary	B-cell_type
immature	I-cell_type
thymocytes	I-cell_type
consequent	O
to	O
T	O
cell	O
receptor	O
(	O
TCR	O
)	O
-mediated	O
ligation	O
.	O

Whereas	O
expression	O
of	O
E2A	O
mRNA	O
and	O
protein	O
are	O
unaltered	O
,	O
Id3	B-RNA
transcripts	I-RNA
are	O
rapidly	O
induced	O
upon	O
signaling	O
from	O
the	O
TCR	B-protein
.	O

Activation	O
of	O
Id3	O
transcription	O
is	O
regulated	O
in	O
a	O
dose-dependent	O
manner	O
by	O
the	O
extracellular	O
signal-regulated	O
kinase	O
(	O
ERK	B-protein
)	O
MAPK	B-protein
module	O
.	O

These	O
observations	O
directly	O
connect	O
the	O
ERK	B-protein
MAPK	B-protein
cascade	O
and	O
HLH	B-protein
proteins	I-protein
in	O
a	O
linear	O
pathway	O
.	O

-DOCSTART-	O

Retinoic	O
acid	O
up-regulates	O
myeloid	O
ICAM-3	O
expression	O
and	O
function	O
in	O
a	O
cell-specific	O
fashion	O
--	O
evidence	O
for	O
retinoid	O
signaling	O
pathways	O
in	O
the	O
mast	B-cell_type
cell	I-cell_type
lineage	I-cell_type
.	O

Investigation	O
of	O
mast	O
cell	O
responsiveness	O
toward	O
retinoic	O
acid	O
(	O
RA	O
)	O
revealed	O
selective	O
promotion	O
of	O
ICAM-3	B-protein
expression	O
in	O
the	O
human	B-cell_line
mast	I-cell_line
cell	I-cell_line
line	I-cell_line
HMC-1	I-cell_line
.	O

This	O
process	O
was	O
dose-	O
and	O
time-dependent	O
and	O
detectable	O
by	O
flow	O
cytometry	O
,	O
Western	O
blot	O
analysis	O
,	O
ELISA	O
,	O
and	O
Northern	O
blot	O
analysis	O
.	O

ICAM-3	B-protein
modulation	O
was	O
found	O
to	O
be	O
cell-type	O
dependent	O
,	O
detectable	O
also	O
for	O
HL-60	B-cell_line
cells	I-cell_line
and	O
monocytes	B-cell_type
but	O
not	O
U-937	B-cell_line
and	O
only	O
weakly	O
for	O
KU812	B-cell_line
cells	I-cell_line
.	O

Terminally	B-cell_type
differentiated	I-cell_type
skin	I-cell_type
mast	I-cell_type
cells	I-cell_type
also	O
failed	O
to	O
up-modulate	O
their	O
ICAM-3	B-protein
,	O
suggesting	O
the	O
requirement	O
for	O
some	O
degree	O
of	O
immaturity	O
for	O
the	O
process	O
.	O

RA-mediated	O
effects	O
on	O
ICAM-1	B-protein
expression	O
,	O
studied	O
in	O
parallel	O
,	O
were	O
clearly	O
distinct	O
from	O
those	O
on	O
ICAM-3	B-protein
.	O

Investigation	O
of	O
retinoid	B-protein
receptor	I-protein
expression	O
,	O
known	O
to	O
mediate	O
intracellular	O
RA	O
signaling	O
,	O
revealed	O
presence	O
of	O
RAR	B-RNA
alpha	I-RNA
,	I-RNA
RAR	I-RNA
gamma	I-RNA
,	I-RNA
RXR	I-RNA
beta	I-RNA
,	I-RNA
and	I-RNA
RXR	I-RNA
gamma	I-RNA
transcripts	I-RNA
in	O
all	O
cell	O
lines	O
studied	O
,	O
and	O
HMC-1	B-cell_line
cells	I-cell_line
were	O
the	O
only	O
line	O
lacking	O
RXR	B-protein
alpha	I-protein
.	O

RAR	B-protein
beta	I-protein
,	O
not	O
expressed	O
at	O
baseline	O
,	O
was	O
induced	O
by	O
RA	O
in	O
a	O
fashion	O
obviously	O
correlating	O
with	O
ICAM-3	B-protein
up-regulation	O
.	O

Increased	O
ICAM-3	B-protein
expression	O
was	O
of	O
functional	O
significance	O
,	O
such	O
that	O
processes	O
stimulated	O
or	O
co-stimulated	O
via	O
ICAM-3	B-protein
(	O
homotypic	O
aggregation	O
,	O
IL-8	O
secretion	O
)	O
were	O
clearly	O
enhanced	O
upon	O
RA	O
pretreatment	O
,	O
suggesting	O
that	O
RA	O
may	O
contribute	O
via	O
hitherto	O
unrecognized	O
pathways	O
to	O
immune	O
function	O
and	O
host	O
defense	O
.	O

-DOCSTART-	O

CD28	B-protein
and	O
T	B-cell_type
cell	I-cell_type
co-stimulation	O
.	O

Over	O
the	O
last	O
decade	O
the	O
concept	O
of	O
T	B-cell_type
cell	I-cell_type
co-stimulation	O
has	O
emerged	O
to	O
take	O
a	O
central	O
role	O
in	O
the	O
process	O
of	O
T	O
cell	O
activation	O
.	O

However	O
,	O
the	O
exact	O
definition	O
of	O
co-stimulation	O
is	O
still	O
unclear	O
.	O

In	O
this	O
review	O
,	O
we	O
re-examine	O
the	O
concept	O
of	O
co-stimulation	O
.	O

We	O
suggest	O
that	O
while	O
co-stimulation	O
is	O
important	O
,	O
there	O
is	O
little	O
evidence	O
to	O
link	O
co-stimulation	O
with	O
T	B-cell_type
cell	I-cell_type
anergy	O
.	O

We	O
then	O
suggest	O
a	O
framework	O
for	O
studying	O
co-stimulation	O
.	O

Focusing	O
on	O
recent	O
advances	O
in	O
our	O
understanding	O
of	O
CD28	B-protein
,	O
we	O
discuss	O
four	O
areas	O
of	O
T	B-cell_type
cell	I-cell_type
activation	O
where	O
co-stimulation	O
may	O
play	O
a	O
role	O
.	O

-DOCSTART-	O

Regulation	O
of	O
activator	B-protein
protein-1	I-protein
and	O
NF-kappa	B-protein
B	I-protein
in	O
CD8+	B-cell_type
T	I-cell_type
cells	I-cell_type
exposed	O
to	O
peripheral	B-protein
self-antigens	I-protein
.	O

The	O
transcriptional	O
events	O
that	O
control	O
T	B-cell_type
cell	I-cell_type
tolerance	O
to	O
peripheral	B-protein
self	I-protein
Ags	I-protein
are	O
still	O
unknown	O
.	O

In	O
this	O
study	O
,	O
we	O
analyzed	O
the	O
regulation	O
of	O
AP-1	B-protein
-	O
and	O
NF-kappa	B-protein
B	I-protein
-mediated	O
transcription	O
during	O
in	O
vivo	O
induction	O
of	O
tolerance	O
to	O
a	O
self	B-protein
Ag	I-protein
expressed	O
exclusively	O
on	O
hepatocytes	B-cell_type
.	O

Naive	B-cell_type
CD8	I-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
Desire	I-cell_type
(	I-cell_type
Des	I-cell_type
)	I-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
T	I-cell_type
cells	I-cell_type
isolated	O
from	O
the	O
Des	O
TCR-transgenic	O
mice	O
that	O
are	O
specific	O
for	O
the	O
H-2K	B-protein
(	I-protein
b	I-protein
)	I-protein
class	I-protein
I	I-protein
Ag	I-protein
were	O
transferred	O
into	O
Alb-K	O
(	O
b	O
)	O
-transgenic	O
mice	O
that	O
express	O
the	O
H-2K	B-protein
(	I-protein
b	I-protein
)	I-protein
Ag	I-protein
on	O
hepatocytes	B-cell_type
only	O
.	O

Tolerance	O
develops	O
in	O
these	O
mice	O
.	O

We	O
found	O
that	O
the	O
self-reactive	B-cell_type
CD8	I-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
Des	I-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
T	I-cell_type
cells	I-cell_type
were	O
transiently	O
activated	O
,	O
then	O
became	O
unresponsive	O
and	O
were	O
further	O
deleted	O
.	O

In	O
contrast	O
to	O
CD8	B-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
Des	I-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
T	I-cell_type
cells	I-cell_type
activated	O
in	O
vivo	O
with	O
APCs	O
,	O
which	O
express	O
high	O
AP-1	B-protein
and	O
high	O
NF-kappa	B-protein
B	I-protein
transcriptional	O
activity	O
,	O
the	O
unresponsive	O
CD8	B-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
Des	I-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
T	I-cell_type
cells	I-cell_type
expressed	O
no	O
AP-1	B-protein
and	O
only	O
weak	O
NF-kappa	B-protein
B	I-protein
transcriptional	O
activity	O
.	O

The	O
differences	O
in	O
NF-kappa	B-protein
B	I-protein
transcriptional	O
activity	O
correlated	O
with	O
the	O
generation	O
of	O
distinct	O
NF-kappa	B-protein
B	I-protein
complexes	I-protein
.	O

Indeed	O
,	O
in	O
vivo	O
primed	B-cell_type
T	I-cell_type
cells	I-cell_type
predominantly	O
express	O
p50/p50	B-protein
and	I-protein
p65/p50	I-protein
dimers	I-protein
,	O
whereas	O
these	O
p50-containing	B-protein
complexes	I-protein
are	O
barely	O
detectable	O
in	O
tolerant	B-cell_type
T	I-cell_type
cells	I-cell_type
that	O
express	O
p65-	B-protein
and	I-protein
c-Rel-containing	I-protein
complexes	I-protein
.	O

These	O
observations	O
suggest	O
that	O
fine	O
regulation	O
of	O
NF-kappa	B-protein
B	I-protein
complex	I-protein
formation	O
may	O
determine	O
T	B-cell_type
cell	I-cell_type
fate	O
.	O

-DOCSTART-	O

Transcriptional	O
regulation	O
in	O
lymphocytes	B-cell_type
.	O

Lymphocytes	B-cell_type
have	O
been	O
used	O
to	O
investigate	O
many	O
cellular	O
processes	O
,	O
including	O
lineage	O
commitment	O
,	O
differentiation	O
,	O
proliferation	O
and	O
apoptosis	O
.	O

The	O
transcription	B-protein
factors	I-protein
that	O
mediate	O
these	O
processes	O
are	O
often	O
expressed	O
broadly	O
in	O
many	O
cell	O
types	O
.	O

The	O
emerging	O
theme	O
is	O
one	O
of	O
cell-type-specific	O
regulation	O
,	O
affecting	O
not	O
only	O
the	O
functional	O
activation	O
of	O
transcription	B-protein
factors	I-protein
but	O
also	O
their	O
access	O
to	O
appropriate	O
regions	O
of	O
DNA	O
.	O

-DOCSTART-	O

Existence	O
of	O
retinoic	B-protein
acid-receptor	I-protein
-independent	O
retinoid	B-protein
X-receptor	I-protein
-dependent	O
pathway	O
in	O
myeloid	B-cell_type
cell	I-cell_type
function	O
.	O

We	O
previously	O
reported	O
that	O
ER-27191	O
(	O
4-	O
[	O
4	O
,	O
5	O
,	O
7	O
,	O
8	O
,	O
9	O
,	O
10-hexahydro-7	O
,	O
7	O
,	O
10	O
,	O
10-tetramethyl-1-	O
(	O
3-pyridylmethyl	O
)	O
anthra	O
[	O
1	O
,	O
2-b	O
]	O
pyrrol-3-yl	O
]	O
benzoic	O
acid	O
)	O
is	O
a	O
potent	O
antagonist	O
of	O
retinoic	B-protein
acid	I-protein
receptor	I-protein
(	O
RAR	B-protein
)	O
,	O
and	O
ER-35795	O
(	O
(	O
2E	O
,	O
4E	O
,	O
6E	O
)	O
-7-	O
[	O
1-	O
(	O
1-methylethyl	O
)	O
-8-chloro-1	O
,	O
2	O
,	O
3	O
,	O
4-tetrahydroquinolin-6-yl	O
]	O
-6-fluoro-3-methyl-2	O
,	O
4	O
,	O
6-nonatrienoic	O
acid	O
)	O
is	O
a	O
novel	O
retinoid	B-protein
X	I-protein
receptor	I-protein
(	O
RXR	B-protein
)	O
-specific	O
agonist	O
.	O

By	O
using	O
these	O
compounds	O
,	O
we	O
investigated	O
whether	O
distinct	O
RAR	B-protein
-dependent	O
and	O
RXR	B-protein
-dependent	O
pathways	O
operate	O
to	O
mediate	O
the	O
diverse	O
activities	O
of	O
retinoids	O
,	O
particularly	O
,	O
the	O
effects	O
of	O
the	O
RXR	B-protein
pathway	O
on	O
cellular	O
function	O
.	O

ER-27191	O
completely	O
antagonized	O
HL60	B-cell_line
cell	I-cell_line
differentiation	O
induced	O
by	O
all-trans-retinoic	O
acid	O
(	O
atRA	O
)	O
.	O

However	O
,	O
the	O
differentiation	O
induced	O
by	O
the	O
ER-35795	O
was	O
not	O
antagonized	O
at	O
all	O
by	O
the	O
RAR	B-protein
antagonist	O
,	O
but	O
was	O
inhibited	O
by	O
an	O
RXR	B-protein
homodimer	I-protein
antagonist	O
(	O
LGD100754	O
,	O
(	O
2E	O
,	O
4E	O
,	O
6Z	O
)	O
-7-	O
(	O
3-n-propoxy-5	O
,	O
6	O
,	O
7	O
,	O
8-tetrahydro-5	O
,	O
5	O
,	O
8	O
,	O
8-tetramethylnaphthalen-2-yl	O
)	O
-3-methylocta-2	O
,	O
4	O
,	O
6-trienoic	O
acid	O
)	O
.	O

Its	O
agonistic	O
action	O
on	O
RXR/RAR	B-protein
heterodimer	I-protein
,	O
on	O
the	O
other	O
hand	O
,	O
was	O
neutralized	O
by	O
the	O
RAR	B-protein
antagonist	O
.	O

During	O
HL60	B-cell_line
cell	I-cell_line
differentiation	O
,	O
atRA	O
induced	O
RARbeta	B-RNA
mRNA	I-RNA
,	O
while	O
the	O
RXR	B-protein
had	O
no	O
effect	O
.	O

Interestingly	O
,	O
a	O
functional	O
RXR	B-protein
-pathway	O
was	O
also	O
seen	O
in	O
lipopolysaccharide-induced	O
inhibition	O
of	O
mouse	B-cell_type
splenocyte	I-cell_type
proliferation	O
.	O

These	O
results	O
strongly	O
suggest	O
the	O
existence	O
of	O
a	O
pharmacological	O
RXR	B-protein
-dependent	O
pathway	O
that	O
is	O
activated	O
by	O
a	O
ligand	O
that	O
can	O
bind	O
to	O
RXR	B-protein
.	O

-DOCSTART-	O

Targeting	O
Src	B-protein
homology	I-protein
2	I-protein
domain-containing	I-protein
tyrosine	I-protein
phosphatase	I-protein
(	O
SHP-1	B-protein
)	O
into	O
lipid	O
rafts	O
inhibits	O
CD3	B-protein
-induced	O
T	B-cell_type
cell	I-cell_type
activation	O
.	O

To	O
study	O
the	O
mechanism	O
by	O
which	O
protein	B-protein
tyrosine	I-protein
phosphatases	I-protein
(	O
PTPs	B-protein
)	O
regulate	O
CD3	B-protein
-induced	O
tyrosine	O
phosphorylation	O
,	O
we	O
investigated	O
the	O
distribution	O
of	O
PTPs	B-protein
in	O
subdomains	O
of	O
plasma	O
membrane	O
.	O

We	O
report	O
here	O
that	O
the	O
bulk	O
PTP	B-protein
activity	O
associated	O
with	O
T	B-cell_type
cell	I-cell_type
membrane	O
is	O
present	O
outside	O
the	O
lipid	O
rafts	O
,	O
as	O
determined	O
by	O
sucrose	O
density	O
gradient	O
sedimentation	O
.	O

In	O
Jurkat	B-cell_line
T	I-cell_line
cells	I-cell_line
,	O
approximately	O
5	O
--	O
10	O
%	O
of	O
Src	B-protein
homology	I-protein
2	I-protein
domain-containing	I-protein
tyrosine	I-protein
phosphatase	I-protein
(	O
SHP-1	B-protein
)	O
is	O
constitutively	O
associated	O
with	O
plasma	O
membrane	O
,	O
and	O
nearly	O
50	O
%	O
of	O
SHP-2	B-protein
is	O
translocated	O
to	O
plasma	O
membrane	O
after	O
vanadate	O
treatment	O
.	O

Similar	O
to	O
transmembrane	B-protein
PTP	I-protein
,	O
CD45	B-protein
,	O
the	O
membrane-associated	O
populations	O
of	O
SHP-1	B-protein
and	O
SHP-2	B-protein
are	O
essentially	O
excluded	O
from	O
lipid	O
rafts	O
,	O
where	O
other	O
signaling	B-protein
molecules	I-protein
such	O
as	O
Lck	B-protein
,	I-protein
linker	I-protein
for	I-protein
activation	I-protein
of	I-protein
T	I-protein
cells	I-protein
,	O
and	O
CD3	B-protein
zeta	I-protein
are	O
enriched	O
.	O

We	O
further	O
demonstrated	O
that	O
CD3	B-protein
-induced	O
tyrosine	O
phosphorylation	O
of	O
these	O
substrates	O
is	O
largely	O
restricted	O
to	O
lipid	O
rafts	O
,	O
unless	O
PTPs	B-protein
are	O
inhibited	O
.	O

It	O
suggests	O
that	O
a	O
restricted	O
partition	O
of	O
PTPs	B-protein
among	O
membrane	O
subdomains	O
may	O
regulate	O
protein	O
tyrosine	O
phosphorylation	O
in	O
T	B-cell_type
cell	I-cell_type
membrane	O
.	O

To	O
test	O
this	O
hypothesis	O
,	O
we	O
targeted	O
SHP-1	B-protein
into	O
lipid	O
rafts	O
by	O
using	O
the	O
N-terminal	B-protein
region	I-protein
of	I-protein
Lck	I-protein
(	O
residues	B-protein
1	I-protein
--	I-protein
14	I-protein
)	O
.	O

The	O
results	O
indicate	O
that	O
the	O
expression	O
of	O
Lck/SHP-1	B-protein
chimera	I-protein
inside	O
lipid	O
rafts	O
profoundly	O
inhibits	O
CD3	B-protein
-induced	O
tyrosine	O
phosphorylation	O
of	O
CD3	B-protein
zeta/epsilon	I-protein
,	O
IL-2	O
generation	O
,	O
and	O
nuclear	O
mobilization	O
of	O
NF-AT	B-protein
.	O

Collectively	O
,	O
these	O
results	O
suggest	O
that	O
the	O
exclusion	O
of	O
PTPs	B-protein
from	O
lipid	O
rafts	O
may	O
be	O
a	O
mechanism	O
that	O
potentiates	O
TCR	B-protein
/	O
CD3	B-protein
activation	O

-DOCSTART-	O

An	O
instructive	O
component	O
in	O
T	B-cell_type
helper	I-cell_type
cell	I-cell_type
type	I-cell_type
2	I-cell_type
(	O
Th2	B-cell_type
)	O
development	O
mediated	O
by	O
GATA-3	B-protein
.	O

Although	O
interleukin	B-protein
(	I-protein
IL	I-protein
)	I-protein
-12	I-protein
and	O
IL-4	B-protein
polarize	O
naive	B-cell_type
CD4	I-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
T	I-cell_type
cells	I-cell_type
toward	O
T	O
helper	O
cell	O
type	O
1	O
(	O
Th1	O
)	O
or	O
Th2	O
phenotypes	O
,	O
it	O
is	O
not	O
known	O
whether	O
cytokines	B-protein
instruct	O
the	O
developmental	O
fate	O
in	O
uncommitted	B-cell_type
progenitors	I-cell_type
or	O
select	O
for	O
outgrowth	O
of	O
cells	O
that	O
have	O
stochastically	O
committed	O
to	O
a	O
particular	O
fate	O
.	O

To	O
distinguish	O
these	O
instructive	O
and	O
selective	O
models	O
,	O
we	O
used	O
surface	O
affinity	O
matrix	O
technology	O
to	O
isolate	O
committed	B-cell_type
progenitors	I-cell_type
based	O
on	O
cytokine	O
secretion	O
phenotype	O
and	O
developed	O
retroviral-based	O
tagging	O
approaches	O
to	O
directly	O
monitor	O
individual	O
progenitor	O
fate	O
decisions	O
at	O
the	O
clonal	O
and	O
population	O
levels	O
.	O

We	O
observe	O
IL-4	B-protein
-dependent	O
redirection	O
of	O
phenotype	O
in	O
cells	O
that	O
have	O
already	O
committed	O
to	O
a	O
non-IL-4-producing	O
fate	O
,	O
inconsistent	O
with	O
predictions	O
of	O
the	O
selective	O
model	O
.	O

Further	O
,	O
retroviral	O
tagging	O
of	O
naive	B-cell_type
progenitors	I-cell_type
with	O
the	O
Th2-specific	B-protein
transcription	I-protein
factor	I-protein
GATA-3	I-protein
provided	O
direct	O
evidence	O
for	O
instructive	O
differentiation	O
,	O
and	O
no	O
evidence	O
for	O
the	O
selective	O
outgrowth	O
of	O
cells	O
committed	O
to	O
either	O
the	O
Th1	O
or	O
Th2	O
fate	O
.	O

These	O
data	O
would	O
seem	O
to	O
exclude	O
selection	O
as	O
an	O
exclusive	O
mechanism	O
in	O
Th1	B-cell_type
/	O
Th2	B-cell_type
differentiation	O
,	O
and	O
support	O
an	O
instructive	O
model	O
of	O
cytokine-driven	O
transcriptional	O
programming	O
of	O
cell	O
fate	O
decisions	O
.	O

-DOCSTART-	O

In	O
vitro-activated	O
human	B-cell_type
lupus	I-cell_type
T	I-cell_type
cells	I-cell_type
express	O
normal	O
estrogen	B-protein
receptor	I-protein
proteins	I-protein
which	O
bind	O
to	O
the	O
estrogen	B-protein
response	I-protein
element	I-protein
.	O

We	O
have	O
shown	O
that	O
estrogen	B-protein
receptor	I-protein
(	O
ERalpha	B-protein
,	O
ERbeta	B-protein
)	O
transcripts	O
are	O
expressed	O
in	O
SLE	B-cell_type
and	I-cell_type
normal	I-cell_type
T	I-cell_type
cells	I-cell_type
.	O

In	O
this	O
study	O
,	O
T	O
cell	O
nuclear	O
extracts	O
from	O
female	O
lupus	O
patients	O
and	O
normal	O
donors	O
were	O
tested	O
for	O
biologically	O
active	O
ER	B-protein
proteins	I-protein
capable	O
of	O
binding	O
to	O
the	O
human	B-DNA
estrogen	I-DNA
response	I-DNA
element	I-DNA
(	O
hERE	B-DNA
)	O
by	O
electrophoretic	O
mobility	O
shift	O
assays	O
.	O

When	O
peripheral	B-cell_type
blood	I-cell_type
T	I-cell_type
cells	I-cell_type
were	O
stimulated	O
with	O
17beta-estradiol	O
(	O
E2	O
)	O
,	O
PMA	O
and	O
ionomycin	O
,	O
two	O
major	O
retarded	O
bands	O
in	O
T	O
cell	O
nuclear	O
extracts	O
exhibited	O
a	O
migration	O
pattern	O
similar	O
to	O
slow	O
migrating	O
protein-ERE	B-protein
complexes	I-protein
in	O
human	O
breast	O
cancer	O
cell	O
extracts	O
.	O

T	B-cell_type
cells	I-cell_type
cultured	O
only	O
with	O
E2	O
did	O
not	O
have	O
these	O
complexes	O
.	O

The	O
formation	O
of	O
the	O
complexes	O
was	O
inhibited	O
by	O
competition	O
with	O
the	O
hERE	O
cold	O
oligonucleotide	O
and	O
partially	O
with	O
anti-ERalpha	B-protein
antibodies	I-protein
.	O

There	O
was	O
no	O
notable	O
difference	O
in	O
the	O
migration	O
pattern	O
of	O
ERE-binding	B-protein
proteins	I-protein
between	O
the	O
SLE	O
and	O
normal	O
T	O
cell	O
extracts	O
.	O

Together	O
,	O
these	O
results	O
suggest	O
that	O
activated	B-cell_type
human	I-cell_type
T	I-cell_type
cells	I-cell_type
,	O
whether	O
lupus-derived	O
or	O
normal-derived	O
,	O
contain	O
biologically	O
active	O
ERalpha	B-protein
proteins	I-protein
.	O

Other	O
factors	O
may	O
be	O
responsible	O
for	O
differential	O
sensitivity	O
of	O
lupus	B-cell_type
T	I-cell_type
cells	I-cell_type
to	O
estrogen	O
.	O

-DOCSTART-	O

Mechanism	O
of	O
the	O
inhibitory	O
effect	O
of	O
protease	O
inhibitor	O
on	O
tumor	B-protein
necrosis	I-protein
factor	I-protein
alpha	I-protein
production	O
of	O
monocytes	B-cell_type
.	O

If	O
the	O
inflammatory	O
response	O
becomes	O
excessive	O
or	O
uncontrolled	O
by	O
some	O
stimuli	O
,	O
inappropriate	O
inflammatory	O
responses	O
occur	O
.	O

Monocytes	B-cell_type
are	O
extremely	O
important	O
cells	O
for	O
regulating	O
the	O
cytokine	O
network	O
and	O
tumor	B-protein
necrosis	I-protein
factor	I-protein
alpha	I-protein
(	O
TNFalpha	B-protein
)	O
and	O
interleukin-	B-protein
(	I-protein
IL	I-protein
)	I-protein
10	I-protein
,	O
which	O
are	O
mainly	O
synthesized	O
by	O
monocytes	B-cell_type
,	O
are	O
representative	O
cytokines	B-protein
that	O
play	O
a	O
central	O
role	O
in	O
the	O
cytokine	O
network	O
.	O

Protease	O
inhibitors	O
such	O
as	O
gabexate	O
mesilate	O
(	O
GM	O
)	O
and	O
ulinastatin	O
(	O
UTI	O
)	O
have	O
been	O
shown	O
to	O
have	O
various	O
beneficial	O
effects	O
by	O
inhibiting	O
the	O
activation	O
of	O
leukocytes	B-cell_type
,	O
but	O
the	O
mechanism	O
for	O
this	O
has	O
yet	O
to	O
be	O
fully	O
elucidated	O
.	O

In	O
this	O
study	O
we	O
investigated	O
the	O
mechanism	O
of	O
the	O
inhibitory	O
effect	O
of	O
protease	O
inhibitors	O
on	O
the	O
proinflammatory	O
cytokine	O
production	O
of	O
lipopolysaccharide-	O
(	O
LPS	O
)	O
stimulated	O
monocytes	B-cell_type
.	O

LPS-stimulated	O
monocytes	B-cell_type
were	O
treated	O
with	O
GM	O
or	O
UTI	O
.	O

The	O
value	O
of	O
TNFalpha	B-protein
and	O
IL-10	B-protein
in	O
the	O
culture	O
medium	O
of	O
monocytes	B-cell_type
was	O
measured	O
and	O
each	O
mRNA	O
expression	O
was	O
assayed	O
.	O

The	O
inhibitory	O
effect	O
of	O
protease	O
inhibitors	O
on	O
the	O
activity	O
of	O
intracellular	O
signal	O
transduction	O
pathways	O
such	O
as	O
protein	B-protein
kinase	I-protein
C	I-protein
(	O
PKC	B-protein
)	O
and	O
nuclear	B-protein
factor	I-protein
kappa	I-protein
B	I-protein
(	O
NFkappaB	B-protein
)	O
were	O
also	O
evaluated	O
.	O

GM	O
decreased	O
the	O
TNFalpha	B-protein
production	O
of	O
LPS-stimulated	O
monocytes	B-cell_type
as	O
shown	O
by	O
the	O
inhibition	O
of	O
mRNA	O
expression	O
and	O
increased	O
the	O
IL-10	B-protein
production	O
of	O
LPS-stimulated	O
monocytes	B-cell_type
.	O

GM	O
also	O
suppressed	O
the	O
NFkappaB	B-protein
activity	O
of	O
LPS-stimulated	O
monocytes	B-cell_type
.	O

UTI	O
decreased	O
the	O
TNFalpha	B-protein
production	O
of	O
LPS-stimulated	O
monocytes	B-cell_type
,	O
but	O
did	O
not	O
inhibit	O
the	O
TNFalpha	B-RNA
mRNA	I-RNA
expression	O
.	O

The	O
present	O
study	O
shows	O
that	O
the	O
inhibitory	O
effect	O
of	O
GM	O
on	O
the	O
TNFalpha	B-protein
production	O
of	O
activated	O
human	B-cell_type
monocytes	I-cell_type
is	O
mediated	O
by	O
the	O
suppression	O
of	O
NFkappaB	B-protein
activation	O
,	O
while	O
the	O
mechanism	O
of	O
UTI	O
inhibiting	O
TNFalpha	B-protein
production	O
of	O
human	B-cell_type
monocytes	I-cell_type
may	O
be	O
due	O
to	O
the	O
inhibition	O
of	O
either	O
the	O
translation	O
or	O
secretion	O
of	O
TNFalpha	B-protein
.	O

-DOCSTART-	O

Homocysteine	O
stimulates	O
the	O
expression	O
of	O
monocyte	B-protein
chemoattractant	I-protein
protein-1	I-protein
in	O
endothelial	B-cell_type
cells	I-cell_type
leading	O
to	O
enhanced	O
monocyte	O
chemotaxis	O
.	O

Hyperhomocysteinemia	O
has	O
been	O
identified	O
as	O
an	O
independent	O
risk	O
factor	O
for	O
atherosclerosis	O
.	O

The	O
infiltration	O
of	O
monocytes	B-cell_type
into	O
the	O
arterial	O
wall	O
is	O
one	O
of	O
the	O
key	O
events	O
during	O
atherogenesis	O
.	O

Monocyte	B-protein
chemoattractant	I-protein
protein-1	I-protein
(	O
MCP-1	B-protein
)	O
is	O
a	O
potent	O
chemokine	B-protein
that	O
stimulates	O
the	O
migration	O
of	O
monocytes	B-cell_type
into	O
the	O
intima	O
of	O
the	O
arterial	O
wall	O
.	O

The	O
mechanism	O
by	O
which	O
increased	O
monocyte	O
infiltration	O
occurs	O
in	O
atherosclerotic	O
lesions	O
in	O
patients	O
with	O
hyperhomocysteinemia	O
has	O
not	O
been	O
delineated	O
.	O

The	O
objective	O
of	O
the	O
present	O
study	O
was	O
to	O
investigate	O
the	O
effect	O
of	O
homocysteine	O
on	O
MCP-1	B-protein
production	O
in	O
endothelial	B-cell_type
cells	I-cell_type
.	O

Cells	O
were	O
incubated	O
with	O
homocysteine	O
.	O

The	O
secretion	O
of	O
MCP-1	B-protein
protein	I-protein
was	O
significantly	O
increased	O
(	O
195	O
%	O
as	O
compared	O
to	O
the	O
control	O
)	O
in	O
cells	O
treated	O
with	O
pathological	O
concentrations	O
of	O
homocysteine	O
.	O

Such	O
effect	O
was	O
accompanied	O
by	O
an	O
increased	O
expression	O
of	O
MCP-1	B-RNA
mRNA	I-RNA
(	O
176	O
%	O
as	O
compared	O
to	O
the	O
control	O
)	O
in	O
endothelial	B-cell_type
cells	I-cell_type
which	O
resulted	O
in	O
enhanced	O
monocyte	O
chemotaxis	O
.	O

The	O
p38	B-protein
MAP	I-protein
kinase	I-protein
as	O
well	O
as	O
other	O
members	O
of	O
the	O
p38	O
MAP	O
kinase	O
pathway	O
,	O
including	O
MKK3	B-protein
,	O
MKK6	B-protein
,	O
ATF-2	B-protein
and	O
Elk-1	B-protein
,	O
were	O
activated	O
in	O
homocysteine-treated	B-cell_type
cells	I-cell_type
.	O

Homocysteine-induced	O
MCP-1	B-protein
expression	O
and	O
subsequent	O
monocyte	O
chemotaxis	O
were	O
blocked	O
by	O
a	O
p38	B-protein
MAP	I-protein
kinase	I-protein
inhibitor	O
(	O
SB203580	O
)	O
suggesting	O
that	O
the	O
p38	B-protein
MAP	I-protein
kinase	I-protein
pathway	O
might	O
be	O
involved	O
in	O
homocysteine-induced	O
MCP-1	B-protein
expression	O
in	O
endothelial	B-cell_type
cells	I-cell_type
.	O

In	O
contrast	O
,	O
staurosporine	O
,	O
a	O
protein	B-protein
kinase	I-protein
C	I-protein
inhibitor	O
,	O
had	O
no	O
effect	O
on	O
homocysteine-induced	O
MCP-1	B-protein
expression	O
.	O

In	O
conclusion	O
,	O
our	O
results	O
indicate	O
that	O
homocysteine	O
stimulates	O
MCP-1	B-protein
expression	O
in	O
endothelial	B-cell_type
cells	I-cell_type
leading	O
to	O
enhanced	O
monocyte	O
chemotaxis	O
.	O

-DOCSTART-	O

Inducible	O
resistance	O
to	O
Fas-mediated	O
apoptosis	O
in	O
B	B-cell_type
cells	I-cell_type
.	O

Apoptosis	O
produced	O
in	O
B	B-cell_type
cells	I-cell_type
through	O
Fas	B-protein
(	O
APO-1	B-protein
,	O
CD95	B-protein
)	O
triggering	O
is	O
regulated	O
by	O
signals	O
derived	O
from	O
other	O
surface	O
receptors	O
:	O
CD40	O
engagement	O
produces	O
upregulation	O
of	O
Fas	B-protein
expression	O
and	O
marked	O
susceptibility	O
to	O
Fas	B-protein
-induced	O
cell	O
death	O
,	O
whereas	O
antigen	O
receptor	O
engagement	O
,	O
or	O
IL-4R	O
engagement	O
,	O
inhibits	O
Fas	B-protein
killing	O
and	O
in	O
so	O
doing	O
induces	O
a	O
state	O
of	O
Fas	B-protein
-resistance	O
,	O
even	O
in	O
otherwise	O
sensitive	O
,	O
CD40	B-protein
-stimulated	O
targets	O
.	O

Surface	B-protein
immunoglobulin	I-protein
and	O
IL-4R	B-protein
utilize	O
at	O
least	O
partially	O
distinct	O
pathways	O
to	O
produce	O
Fas	B-protein
-resistance	O
that	O
differentially	O
depend	O
on	O
PKC	B-protein
and	O
STAT6	B-protein
,	O
respectively	O
.	O

Further	O
,	O
surface	O
immunoglobulin	O
signaling	O
for	O
inducible	O
Fas	B-protein
-resistance	O
bypasses	O
Btk	B-protein
,	O
requires	O
NF-kappaB	B-protein
,	O
and	O
entails	O
new	O
macromolecular	O
synthesis	O
.	O

Terminal	O
effectors	O
of	O
B	B-cell_type
cell	I-cell_type
Fas	B-protein
-resistance	O
include	O
the	O
known	O
anti-apoptotic	B-protein
gene	I-protein
products	I-protein
,	O
Bcl-xL	B-protein
and	O
FLIP	B-protein
,	O
and	O
a	O
novel	O
anti-apoptotic	B-DNA
gene	I-DNA
that	O
encodes	O
FAIM	B-protein
(	O
Fas	B-protein
Apoptosis	I-protein
Inhibitory	I-protein
Molecule	I-protein
)	O
.	O

faim	B-protein
was	O
identified	O
by	O
differential	O
display	O
and	O
exists	O
in	O
two	O
alternatively	O
spliced	O
forms	O
;	O
faim-S	B-protein
is	O
broadly	O
expressed	O
,	O
but	O
faim-L	O
expression	O
is	O
tissue-specific	O
.	O

The	O
FAIM	B-DNA
sequence	I-DNA
is	O
highly	O
evolu-	O
tionarily	O
conserved	O
,	O
suggesting	O
an	O
important	O
role	O
for	O
this	O
molecule	O
throughout	O
phylogeny	O
.	O

Inducible	O
resistance	O
to	O
Fas	B-protein
killing	O
is	O
hypothesized	O
to	O
protect	O
foreign	O
antigen-specific	O
B	B-cell_type
cells	I-cell_type
during	O
potentially	O
hazardous	O
interactions	O
with	O
FasL-bearing	B-cell_type
T	I-cell_type
cells	I-cell_type
,	O
whereas	O
autoreactive	B-cell_type
B	I-cell_type
cells	I-cell_type
fail	O
to	O
become	O
Fas	B-protein
-resistant	O
and	O
are	O
deleted	O
via	O
Fas	B-protein
-dependent	O
cytotoxicity	O
.	O

Inadvertent	O
or	O
aberrant	O
acquisition	O
of	O
Fas	B-protein
-resistance	O
may	O
permit	O
autoreactive	B-cell_type
B	I-cell_type
cells	I-cell_type
to	O
escape	O
Fas	B-protein
deletion	O
,	O
and	O
malignant	B-cell_type
lymphocytes	I-cell_type
to	O
impede	O
anti-tumor	O
immunity	O
.	O

-DOCSTART-	O

Stromal	B-protein
cell-derived	I-protein
factor	I-protein
1	I-protein
alpha	I-protein
-induced	O
chemotaxis	O
in	O
T	B-cell_type
cells	I-cell_type
is	O
mediated	O
by	O
nitric	O
oxide	O
signaling	O
pathways	O
.	O

Stromal	B-protein
cell-derived	I-protein
factor	I-protein
1	I-protein
alpha	I-protein
(	O
SDF1	B-protein
alpha	I-protein
)	O
and	O
its	O
cognate	O
chemokine	B-protein
receptor	I-protein
CXCR4	I-protein
act	O
as	O
potent	O
chemoattractants	O
and	O
regulate	O
trafficking	O
and	O
homing	O
of	O
hematopoietic	B-cell_type
progenitor	I-cell_type
cells	I-cell_type
and	O
lymphocytes	B-cell_type
.	O

However	O
,	O
the	O
molecular	O
mechanisms	O
regulating	O
SDF1	B-protein
alpha	I-protein
-driven	O
cell	O
migration	O
are	O
not	O
well	O
defined	O
.	O

In	O
this	O
study	O
,	O
we	O
have	O
explored	O
the	O
roles	O
of	O
the	O
second	O
messenger	O
NO	O
and	O
the	O
transcription	B-protein
factor	I-protein
NF-kappa	I-protein
B	I-protein
in	O
SDF1	B-protein
alpha	I-protein
-induced	O
T	O
cell	O
migration	O
.	O

SDF1	B-protein
alpha	I-protein
treatment	O
of	O
Jurkat	B-cell_line
T	I-cell_line
cells	I-cell_line
increased	O
the	O
activity	O
of	O
NO	B-protein
synthase	I-protein
,	O
which	O
catalyzes	O
the	O
generation	O
of	O
NO	O
.	O

We	O
observed	O
that	O
pretreatment	O
of	O
Jurkat	B-cell_line
cells	I-cell_line
or	O
activated	O
PBLs	B-cell_type
with	O
several	O
NO	O
donors	O
significantly	O
enhanced	O
the	O
SDF1	B-protein
alpha	I-protein
-induced	O
migration	O
,	O
whereas	O
various	O
inhibitors	O
of	O
NO	B-protein
synthase	I-protein
markedly	O
abrogated	O
the	O
chemotactic	O
response	O
in	O
a	O
concentration-dependent	O
manner	O
.	O

Furthermore	O
,	O
we	O
observed	O
that	O
inhibitors	O
of	O
the	O
transcription	B-protein
factor	I-protein
NF-kappa	I-protein
B	I-protein
,	O
which	O
is	O
linked	O
to	O
NO	O
signaling	O
pathways	O
,	O
also	O
significantly	O
blocked	O
the	O
SDF1	B-protein
alpha	I-protein
-induced	O
chemotactic	O
response	O
.	O

However	O
,	O
these	O
compounds	O
did	O
not	O
have	O
a	O
significant	O
effect	O
on	O
SDF1	B-protein
alpha	I-protein
-induced	O
mitogen-activated	O
protein	O
kinase	O
activity	O
.	O

In	O
addition	O
,	O
the	O
MAP/Erk	O
kinase	O
kinase	O
inhibitor	O
PD98059	O
did	O
not	O
abrogate	O
SDF1	B-protein
alpha	I-protein
-induced	O
chemotaxis	O
.	O

AKT	B-protein
,	O
which	O
has	O
been	O
shown	O
to	O
mediate	O
NO	O
production	O
,	O
was	O
also	O
phosphorylated	O
upon	O
SDF1	B-protein
alpha	I-protein
stimulation	O
.	O

These	O
studies	O
suggest	O
that	O
NO-related	O
signaling	O
pathways	O
may	O
mediate	O
SDF1	B-protein
alpha	I-protein
-induced	O
chemotaxis	O
,	O
but	O
not	O
mitogen-activated	B-protein
protein	I-protein
kinase	I-protein
activation	O
.	O

-DOCSTART-	O

The	O
lack	O
of	O
NF-kappa	O
B	O
transactivation	O
and	O
PKC	O
epsilon	O
expression	O
in	O
CD4	B-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
CD8	I-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
thymocytes	I-cell_type
correlates	O
with	O
negative	O
selection	O
.	O

Deletion	O
of	O
autoreactive	B-cell_type
thymocytes	I-cell_type
at	O
the	O
DP	O
stage	O
is	O
the	O
basis	O
for	O
tolerance	O
to	O
thymus-expressed	B-protein
self	I-protein
antigens	I-protein
.	O

In	O
this	O
study	O
we	O
investigated	O
whether	O
distinct	O
signalling	O
pathways	O
are	O
induced	O
in	O
DP	B-cell_type
thymocytes	I-cell_type
as	O
compared	O
to	O
mature	B-cell_type
T	I-cell_type
cells	I-cell_type
upon	O
stimulation	O
with	O
antigen	O
.	O

Using	O
triple	O
transgenic	O
mice	O
expressing	O
a	O
TCR	B-DNA
transgene	I-DNA
,	O
dominant	B-protein
negative	I-protein
ras/Mek	I-protein
proteins	I-protein
and	O
a	O
reporter	B-DNA
gene	I-DNA
construct	I-DNA
with	O
AP-1	B-DNA
or	I-DNA
NF-kappa	I-DNA
B	I-DNA
binding	I-DNA
sites	I-DNA
,	O
we	O
showed	O
a	O
complete	O
lack	O
of	O
transcriptional	O
activity	O
of	O
NF-kappa	B-protein
B	I-protein
but	O
not	O
AP-1	B-protein
in	O
DP	B-cell_type
thymocytes	I-cell_type
,	O
whereas	O
both	O
were	O
transcriptionally	O
active	O
in	O
mature	B-cell_type
T	I-cell_type
cells	I-cell_type
after	O
antigenic	O
stimulation	O
.	O

Lack	O
of	O
NF-kappa	B-protein
B	I-protein
induction	O
correlated	O
with	O
increased	O
death	O
in	O
response	O
to	O
antigen	B-protein
.	O

AP-1	B-protein
induction	O
was	O
dependent	O
on	O
the	O
integrity	O
of	O
the	O
ras/Mek	O
pathway	O
indicating	O
that	O
this	O
pathway	O
was	O
activated	O
in	O
the	O
DP	B-cell_type
thymocytes	I-cell_type
.	O

In	O
contrast	O
,	O
we	O
found	O
a	O
complete	O
lack	O
of	O
constitutive	O
expression	O
of	O
the	O
epsilon	B-protein
isoform	I-protein
of	I-protein
Protein	I-protein
Kinase	I-protein
C	I-protein
(	O
PKC	B-protein
)	O
in	O
DP	B-cell_type
thymocytes	I-cell_type
,	O
although	O
it	O
was	O
present	O
in	O
mature	B-cell_type
thymocytes	I-cell_type
and	O
peripheral	B-cell_type
T	I-cell_type
cells	I-cell_type
.	O

Taken	O
together	O
the	O
results	O
suggest	O
that	O
the	O
lack	O
of	O
PKC	B-protein
epsilon	I-protein
in	O
DP	B-cell_type
thymocytes	I-cell_type
could	O
lead	O
to	O
the	O
absence	O
of	O
NF-kappa	B-protein
B	I-protein
activity	O
after	O
antigenic	O
stimulation	O
contributing	O
to	O
negative	O
selection	O
.	O

Cell	O
Death	O
and	O
Differentiation	O
(	O
2000	O
)	O
7	O
,	O
1253	O
-	O
1262	O
.	O

-DOCSTART-	O

CD2	O
stimulation	O
leads	O
to	O
the	O
delayed	O
and	O
prolonged	O
activation	O
of	O
STAT1	B-protein
in	O
T	B-cell_type
cells	I-cell_type
but	O
not	O
NK	B-cell_type
cells	I-cell_type
.	O

OBJECTIVE	O
:	O
T	B-cell_type
lymphocytes	I-cell_type
can	O
be	O
activated	O
by	O
soluble	O
factors	O
such	O
as	O
cytokines	B-protein
or	O
through	O
direct	O
cell-cell	O
interactions	O
.	O

Although	O
cytokine	B-protein
receptors	I-protein
are	O
known	O
to	O
signal	O
through	O
STAT	B-protein
family	I-protein
transcription	I-protein
factors	I-protein
,	O
the	O
mechanisms	O
by	O
which	O
other	O
cell-surface	B-protein
molecules	I-protein
,	O
such	O
as	O
CD2	B-protein
,	O
transduce	O
signals	O
is	O
unclear	O
.	O

The	O
goal	O
of	O
this	O
study	O
was	O
to	O
determine	O
whether	O
stimulation	O
of	O
T	B-cell_type
cells	I-cell_type
through	O
CD2	B-protein
recapitulates	O
aspects	O
of	O
cytokine	B-protein
-induced	O
T-cell	O
activation	O
by	O
use	O
of	O
STAT	B-protein
transcription	I-protein
factors	I-protein
.	O

MATERIALS	O
AND	O
METHODS	O
:	O
T	B-cell_type
cells	I-cell_type
were	O
treated	O
with	O
anti-CD2	B-protein
antibodies	I-protein
or	O
cells	O
bearing	O
the	O
natural	O
CD2	B-protein
ligand	I-protein
CD58	I-protein
,	O
after	O
which	O
signaling	O
through	O
STAT	B-protein
transcription	I-protein
factors	I-protein
was	O
assessed	O
.	O

RESULTS	O
:	O
Stimulation	O
of	O
CD2	B-protein
on	O
primary	B-cell_type
T	I-cell_type
lymphocytes	I-cell_type
leads	O
to	O
the	O
tyrosine	O
phosphorylation	O
,	O
nuclear	O
translocation	O
,	O
and	O
DNA	O
binding	O
of	O
STAT1	B-protein
.	O

In	O
contrast	O
to	O
stimulation	O
by	O
cytokines	B-protein
,	O
the	O
activation	O
of	O
STAT1	B-protein
in	O
response	O
to	O
CD2	O
ligation	O
is	O
delayed	O
and	O
does	O
not	O
involve	O
Jak	B-protein
kinases	I-protein
.	O

Furthermore	O
,	O
while	O
STAT	O
phosphorylation	O
induced	O
by	O
cytokines	B-protein
is	O
generally	O
transient	O
,	O
STAT1	O
phosphorylation	O
following	O
CD2	O
stimulation	O
persists	O
for	O
a	O
period	O
of	O
days	O
.	O

Transcription	O
of	O
key	O
target	O
genes	O
such	O
as	O
IRF1	B-DNA
and	O
c-fos	B-DNA
proceeds	O
with	O
delayed	O
kinetics	O
following	O
CD2	O
stimulation	O
,	O
suggesting	O
that	O
this	O
unique	O
pattern	O
of	O
STAT	O
activation	O
may	O
lead	O
to	O
a	O
distinct	O
cellular	O
response	O
following	O
CD2	O
ligation	O
.	O

This	O
pathway	O
appears	O
to	O
be	O
restricted	O
to	O
T	B-cell_type
cells	I-cell_type
,	O
as	O
stimulation	O
of	O
CD2	B-protein
on	O
NK	B-cell_type
cells	I-cell_type
does	O
not	O
lead	O
to	O
STAT1	O
activation	O
.	O

CONCLUSION	O
:	O
Stimulation	O
of	O
T	B-cell_type
cells	I-cell_type
through	O
cell-surface	B-protein
molecules	I-protein
such	O
as	O
CD2	B-protein
involves	O
activation	O
of	O
STAT	B-protein
transcription	I-protein
factors	I-protein
,	O
thus	O
recapitulating	O
elements	O
of	O
cytokine	O
signaling	O
.	O

-DOCSTART-	O

NFATc1	B-protein
and	O
NFATc2	B-protein
together	O
control	O
both	O
T	O
and	O
B	O
cell	O
activation	O
and	O
differentiation	O
.	O

NFAT	B-protein
transcription	I-protein
factors	I-protein
play	O
critical	O
roles	O
in	O
gene	O
transcription	O
during	O
immune	O
responses	O
.	O

To	O
investigate	O
further	O
the	O
two	O
most	O
prominent	O
NFAT	B-protein
family	I-protein
members	I-protein
,	O
NFATc1	B-protein
and	O
NFATc2	B-protein
,	O
we	O
generated	O
mice	O
bearing	O
lymphoid	O
systems	O
devoid	O
of	O
both	O
.	O

Doubly	O
deficient	O
T	B-cell_type
cells	I-cell_type
displayed	O
cell	O
surface	O
markers	O
of	O
activation	O
yet	O
were	O
significantly	O
deficient	O
in	O
the	O
development	O
of	O
multiple	O
effector	O
functions	O
,	O
including	O
Th	B-protein
cytokine	I-protein
production	O
,	O
surface	B-protein
effector	I-protein
molecule	I-protein
expression	O
,	O
and	O
cytolytic	O
activity	O
.	O

Nevertheless	O
,	O
doubly	B-cell_type
deficient	I-cell_type
B	I-cell_type
cells	I-cell_type
were	O
hyperactivated	O
,	O
as	O
evidenced	O
by	O
extremely	O
elevated	O
serum	O
IgG1	B-protein
and	O
IgE	B-protein
,	O
as	O
well	O
as	O
plasma	B-cell_type
cell	I-cell_type
expansion	O
and	O
infiltration	O
of	O
end	O
organs	O
.	O

Thus	O
,	O
in	O
T	B-cell_type
cells	I-cell_type
,	O
NFATc1	B-protein
and	O
NFATc2	B-protein
are	O
dispensable	O
for	O
inflammatory	O
reactivity	O
but	O
are	O
required	O
for	O
effector	O
differentiation	O
,	O
while	O
in	O
B	B-cell_type
cells	I-cell_type
,	O
NFATs	B-protein
regulate	O
both	O
normal	O
homeostasis	O
and	O
differentiation	O
.	O

-DOCSTART-	O

Epstein-barr	O
virus	O
immediate-early	B-protein
protein	I-protein
BZLF1	I-protein
is	O
SUMO-1	O
modified	O
and	O
disrupts	O
promyelocytic	O
leukemia	O
bodies	O
.	O

Although	O
the	O
immediate-early	B-protein
proteins	I-protein
of	O
both	O
herpes	O
simplex	O
virus	O
(	O
HSV	O
)	O
and	O
cytomegalovirus	O
(	O
CMV	O
)	O
are	O
known	O
to	O
modify	O
promyelocytic	O
leukemia	O
(	O
PML	O
)	O
(	O
ND10	O
)	O
bodies	O
in	O
the	O
nucleus	O
of	O
the	O
host	O
cell	O
,	O
it	O
has	O
been	O
unclear	O
whether	O
lytic	O
infection	O
with	O
gamma	O
herpesviruses	O
induces	O
a	O
similar	O
effect	O
.	O

The	O
PML	B-protein
protein	I-protein
is	O
induced	O
by	O
interferon	B-protein
,	O
involved	O
in	O
major	B-protein
histocompatibility	I-protein
complex	I-protein
class	I-protein
I	I-protein
presentation	O
,	O
and	O
necessary	O
for	O
certain	O
types	O
of	O
apoptosis	O
.	O

Therefore	O
,	O
it	O
is	O
likely	O
that	O
PML	O
bodies	O
function	O
in	O
an	O
antiviral	O
capacity	O
.	O

SUMO-1	O
modification	O
of	O
PML	B-protein
is	O
known	O
to	O
be	O
required	O
for	O
the	O
formation	O
of	O
PML	O
bodies	O
.	O

To	O
examine	O
whether	O
Epstein-Barr	O
virus	O
(	O
EBV	O
)	O
lytic	O
replication	O
interferes	O
with	O
PML	O
bodies	O
,	O
we	O
expressed	O
the	O
EBV	B-DNA
immediate-early	I-DNA
genes	I-DNA
BZLF1	B-DNA
(	I-DNA
Z	I-DNA
)	I-DNA
and	O
BRLF1	B-DNA
(	I-DNA
R	I-DNA
)	I-DNA
in	O
EBV-positive	B-cell_line
cell	I-cell_line
lines	I-cell_line
and	O
examined	O
PML	B-protein
localization	O
.	O

Both	O
Z	O
and	O
R	O
expression	O
resulted	O
in	O
PML	B-protein
dispersion	O
in	O
EBV-positive	B-cell_type
cells	I-cell_type
.	O

Z	O
but	O
not	O
R	O
expression	O
is	O
sufficient	O
to	O
disrupt	O
PML	O
bodies	O
in	O
EBV-negative	B-cell_line
cell	I-cell_line
lines	I-cell_line
.	O

We	O
show	O
that	O
dispersion	O
of	O
PML	O
bodies	O
by	O
Z	B-DNA
requires	O
a	O
portion	O
of	O
the	O
transcriptional	B-DNA
activation	I-DNA
domain	I-DNA
of	O
Z	B-DNA
but	O
not	O
the	O
DNA-binding	O
function	O
.	O

As	O
was	O
previously	O
reported	O
for	O
the	O
HSV-1	B-protein
ICP0	I-protein
and	I-protein
CMV	I-protein
IE1	I-protein
proteins	I-protein
,	O
Z	B-DNA
reduces	O
the	O
amount	O
of	O
SUMO-1-modified	B-protein
PML	I-protein
.	O

We	O
also	O
found	O
that	O
Z	B-DNA
itself	O
is	O
SUMO-1	O
modified	O
(	O
through	O
amino	O
acid	O
12	O
)	O
and	O
that	O
Z	B-DNA
competes	O
with	O
PML	B-protein
for	O
limiting	O
amounts	O
of	O
SUMO-1	B-protein
.	O

These	O
results	O
suggest	O
that	O
disruption	O
of	O
PML	O
bodies	O
is	O
important	O
for	O
efficient	O
lytic	O
replication	O
of	O
EBV	O
.	O

Furthermore	O
,	O
Z	B-DNA
may	O
potentially	O
alter	O
the	O
function	O
of	O
a	O
variety	O
of	O
cellular	O
proteins	O
by	O
inhibiting	O
SUMO-1	O
modification	O

-DOCSTART-	O

Suppression	O
of	O
nuclear	B-protein
factor-kappaB	I-protein
and	O
stimulation	O
of	O
inhibitor	B-protein
kappaB	I-protein
by	O
troglitazone	O
:	O
evidence	O
for	O
an	O
anti-inflammatory	O
effect	O
and	O
a	O
potential	O
antiatherosclerotic	O
effect	O
in	O
the	O
obese	O
.	O

To	O
elucidate	O
whether	O
troglitazone	O
exerts	O
an	O
antiinflammatory	O
effect	O
in	O
humans	O
,	O
in	O
vivo	O
,	O
we	O
investigated	O
the	O
suppression	O
of	O
nuclear	B-protein
factor	I-protein
kappaB	I-protein
(	O
NFkappaB	B-protein
)	O
in	O
mononuclear	B-cell_type
cells	I-cell_type
(	I-cell_type
MNC	I-cell_type
)	I-cell_type
by	O
this	O
drug	O
.	O

We	O
measured	O
intranuclear	B-protein
NFkappaB	I-protein
,	O
total	O
cellular	B-protein
NFkappaB	I-protein
,	O
inhibitor	B-protein
kappaB	I-protein
(	I-protein
IkappaB	I-protein
)	I-protein
alpha	I-protein
,	O
reactive	O
oxygen	O
species	O
(	O
ROS	O
)	O
generation	O
,	O
and	O
p47	B-protein
(	I-protein
phox	I-protein
)	I-protein
subunit	I-protein
(	O
a	O
key	O
component	O
protein	O
of	O
nicotinamide	B-protein
adenine	I-protein
dinucleotide	I-protein
phosphate	I-protein
oxidase	I-protein
)	O
in	O
MNC	B-cell_type
.	O

Plasma	B-protein
tumor	I-protein
necrosis	I-protein
factor	I-protein
(	I-protein
TNF	I-protein
)	I-protein
-alpha	I-protein
,	O
soluble	B-protein
intercellular	I-protein
adhesion	I-protein
molecule-1	I-protein
(	O
sICAM-1	B-protein
)	O
,	O
monocyte	B-protein
chemoattractant	I-protein
protein-1	I-protein
(	I-protein
MCP-1	I-protein
)	I-protein
,	O
plasminogen	B-protein
activator	I-protein
inhibitor	I-protein
type	I-protein
1	I-protein
(	O
PAI-1	B-protein
)	O
,	O
C-reactive	B-protein
protein	I-protein
(	I-protein
CRP	I-protein
)	I-protein
,	O
and	O
interleukin	B-protein
(	I-protein
IL	I-protein
)	I-protein
-10	I-protein
(	O
antiinflammatory	B-protein
cytokine	I-protein
)	O
concentrations	O
were	O
also	O
measured	O
as	O
mediators	O
of	O
inflammatory	O
activity	O
that	O
are	O
regulated	O
by	O
the	O
proinflammatory	B-protein
transcription	I-protein
factor	I-protein
NFkappaB	I-protein
.	O

Seven	O
nondiabetic	O
obese	O
patients	O
were	O
given	O
400	O
mg	O
troglitazone	O
daily	O
for	O
4	O
weeks	O
.	O

Blood	O
samples	O
were	O
collected	O
before	O
and	O
at	O
weekly	O
intervals	O
thereafter	O
.	O

MNC	B-cell_type
were	O
separated	O
;	O
and	O
the	O
levels	O
of	O
intranuclear	B-protein
NFkappaB	I-protein
,	O
total	O
cellular	B-protein
NFkappaB	I-protein
,	O
IkappaBalpha	B-protein
,	O
and	O
p47	B-protein
(	I-protein
phox	I-protein
)	I-protein
subunit	I-protein
and	O
ROS	O
generation	O
were	O
determined	O
.	O

Plasma	O
was	O
used	O
to	O
measure	O
insulin	O
glucose	O
,	O
TNFalpha	B-protein
,	O
sICAM	B-protein
,	O
MCP-1	B-protein
,	O
PAI-1	B-protein
,	O
CRP	B-protein
,	O
and	O
IL-10	B-protein
.	O

Plasma	O
insulin	O
concentrations	O
fell	O
significantly	O
at	O
week	O
1	O
,	O
from	O
31.2	O
+/-	O
29.1	O
to	O
14.2	O
+/-	O
11.4	O
mU/L	O
(	O
P	O
<	O
0.01	O
)	O
and	O
remained	O
low	O
throughout	O
4	O
weeks	O
.	O

Plasma	O
glucose	O
concentrations	O
did	O
not	O
alter	O
significantly	O
.	O

There	O
was	O
a	O
fall	O
in	O
intranuclear	B-protein
NFkappaB	I-protein
,	O
total	O
cellular	B-protein
NFkappaB	I-protein
,	O
and	O
p47	B-protein
(	I-protein
phox	I-protein
)	I-protein
subunit	I-protein
,	O
with	O
an	O
increase	O
in	O
cellular	O
IkappaBalpha	B-protein
at	O
week	O
2	O
,	O
which	O
persisted	O
until	O
week	O
4	O
.	O

There	O
was	O
a	O
parallel	O
fall	O
in	O
ROS	O
generation	O
by	O
MNC	B-cell_type
at	O
week	O
1	O
;	O
this	O
progressed	O
and	O
persisted	O
until	O
week	O
4	O
(	O
P	O
<	O
0.001	O
)	O
.	O

Plasma	O
TNF-alpha	B-protein
,	O
sICAM-1	B-protein
,	O
MCP-1	B-protein
,	O
and	O
PAI-1	B-protein
concentrations	O
fell	O
significantly	O
at	O
week	O
4	O
.	O

Plasma	O
IL-10	B-protein
concentration	O
increased	O
significantly	O
,	O
whereas	O
plasma	B-protein
CRP	I-protein
concentrations	O
decreased	O
.	O

We	O
conclude	O
that	O
troglitazone	O
has	O
an	O
antiinflammatory	O
action	O
that	O
may	O
contribute	O
to	O
its	O
putative	O
antiatherosclerotic	O
effects	O
.	O

-DOCSTART-	O

Tyrosine	O
phosphorylation-dependent	O
activation	O
of	O
NF-kappa	B-protein
B	I-protein
.	O

Requirement	O
for	O
p56	B-protein
LCK	I-protein
and	I-protein
ZAP-70	I-protein
protein	I-protein
tyrosine	I-protein
kinases	I-protein
.	O

Phosphorylation	O
of	O
the	O
N-terminal	B-protein
domain	I-protein
of	O
I	B-protein
kappa	I-protein
B	I-protein
inhibitory	I-protein
subunits	I-protein
induces	O
activation	O
of	O
the	O
transcription	B-protein
factor	I-protein
NF-kappa	I-protein
B	I-protein
.	O

Although	O
serine	O
phosphorylation	O
has	O
been	O
shown	O
to	O
induce	O
ubiquitination	O
and	O
subsequent	O
proteasome-mediated	O
degradation	O
of	O
I	B-protein
kappa	I-protein
B-alpha	I-protein
,	O
little	O
is	O
known	O
about	O
the	O
mechanisms	O
that	O
lead	O
to	O
release	O
of	O
active	O
NF-kappa	B-protein
B	I-protein
in	O
T	B-cell_type
cells	I-cell_type
as	O
a	O
consequence	O
of	O
tyrosine	O
phosphorylation	O
of	O
I	B-protein
kappa	I-protein
B-alpha	I-protein
[	O
Imbert	O
,	O
V.	O
,	O
Rupec	O
,	O
R.A.	O
,	O
Livolsi	O
,	O
A.	O
,	O
Pahl	O
,	O
H.L.	O
,	O
Traenckner	O
,	O
B.M.	O
,	O
Mueller-Dieckmann	O
,	O
C.	O
,	O
Farahifar	O
,	O
D.	O
,	O
Rossi	O
,	O
B.	O
,	O
Auberger	O
,	O
P.	O
,	O
Baeuerle	O
,	O
P.	O
&	O
Peyron	O
,	O
J.F.	O
(	O
1996	O
)	O
Cell	O
86	O
,	O
787	O
--	O
798	O
]	O
.	O

The	O
involvement	O
of	O
the	O
tyrosine	B-protein
kinases	I-protein
p56	B-protein
(	I-protein
lck	I-protein
)	I-protein
and	O
ZAP-70	B-protein
in	O
this	O
reaction	O
is	O
demonstrated	O
here	O
using	O
specific	O
pharmacological	O
inhibitors	O
and	O
Jurkat	B-cell_line
mutants	I-cell_line
unable	O
to	O
express	O
these	O
kinases	B-protein
.	O

Although	O
the	O
inhibitors	O
prevented	O
both	O
pervanadate-induced	O
phosphorylation	O
of	O
I	B-protein
kappa	I-protein
B-alpha	I-protein
on	O
Tyr42	O
and	O
NF-kappa	B-protein
B	I-protein
activation	O
,	O
we	O
observed	O
that	O
,	O
in	O
p56	B-cell_line
(	I-cell_line
lck	I-cell_line
)	I-cell_line
-deficient	I-cell_line
Jurkat	I-cell_line
mutants	I-cell_line
,	O
NF-kappa	B-protein
B	I-protein
could	O
still	O
associate	O
with	O
I	B-protein
kappa	I-protein
B-alpha	I-protein
despite	O
phosphorylation	O
on	O
Tyr42	O
.	O

Furthermore	O
,	O
the	O
SH2	B-protein
domain	I-protein
of	O
p56	B-protein
(	I-protein
lck	I-protein
)	I-protein
appeared	O
to	O
be	O
required	O
for	O
pervanadate-induced	O
NF-kappa	B-protein
B	I-protein
activation	O
but	O
not	O
for	O
Tyr42	O
phosphorylation	O
.	O

These	O
results	O
show	O
that	O
p56	B-protein
(	I-protein
lck	I-protein
)	I-protein
and	O
ZAP-70	B-protein
are	O
key	O
components	O
of	O
the	O
signaling	O
pathway	O
that	O
leads	O
to	O
phosphotyrosine-dependent	O
NF-kappa	B-protein
B	I-protein
activation	O
in	O
T	B-cell_type
cells	I-cell_type
and	O
confirm	O
that	O
tyrosine	B-protein
kinases	I-protein
must	O
control	O
at	O
least	O
two	O
different	O
steps	O
to	O
induce	O
activation	O
of	O
NF-kappa	B-protein
B	I-protein
.	O

Finally	O
,	O
we	O
show	O
that	O
H	O
(	O
2	O
)	O
O	O
(	O
2	O
)	O
,	O
which	O
stimulates	O
p56	B-protein
(	I-protein
lck	I-protein
)	I-protein
and	O
ZAP-70	B-protein
in	O
T	B-cell_type
cells	I-cell_type
,	O
is	O
an	O
activator	O
of	O
NF-kappa	B-protein
B	I-protein
through	O
tyrosine	O
phosphorylation	O
of	O
I	B-protein
kappa	I-protein
B-alpha	I-protein
.	O

-DOCSTART-	O

Specific	O
missense	O
mutations	O
in	O
NEMO	B-DNA
result	O
in	O
hyper-IgM	O
syndrome	O
with	O
hypohydrotic	O
ectodermal	O
dysplasia	O
.	O

The	O
gene	O
that	O
encodes	O
nuclear	B-DNA
factor	I-DNA
kappaB	I-DNA
(	I-DNA
NF-kappaB	I-DNA
)	I-DNA
essential	I-DNA
modulator	I-DNA
(	O
or	O
NEMO	B-DNA
,	O
also	O
known	O
as	O
IKKgamma	B-DNA
)	O
is	O
required	O
for	O
activation	O
of	O
the	O
transcription	B-protein
factor	I-protein
NF-kappaB	I-protein
.	O

We	O
describe	O
mutations	O
in	O
the	O
putative	B-protein
zinc-finger	I-protein
domain	I-protein
of	O
NEMO	B-protein
that	O
result	O
in	O
an	O
X-linked	O
primary	O
immunodeficiency	O
characterized	O
by	O
hyper-IgM	O
syndrome	O
and	O
hypohydrotic	O
ectodermal	O
dysplasia	O
(	O
XHM-ED	O
)	O
.	O

These	O
mutations	O
prevent	O
CD40	B-protein
ligand	I-protein
(	O
CD40L	B-protein
)	O
-mediated	O
degradation	O
of	O
inhibitor	O
of	O
NF-kappaB	B-protein
alpha	I-protein
(	O
IkappaB-alpha	B-protein
)	O
and	O
account	O
for	O
the	O
following	O
observations	O
:	O
B	B-cell_type
cells	I-cell_type
from	O
XHM-ED	O
patients	O
are	O
unable	O
to	O
undergo	O
immunoglobulin	O
class-switch	O
recombination	O
and	O
antigen-presenting	B-cell_type
cells	I-cell_type
(	O
APCs	B-cell_type
)	O
are	O
unable	O
to	O
synthesize	O
the	O
NF-kappaB-regulated	B-protein
cytokines	I-protein
interleukin	B-protein
12	I-protein
(	O
IL-12	B-protein
)	O
or	O
tumor	B-protein
necrosis	I-protein
factor	I-protein
alpha	I-protein
(	O
TNF-alpha	B-protein
)	O
when	O
stimulated	O
with	O
CD40L	B-protein
.	O

Nevertheless	O
,	O
innate	O
immunity	O
is	O
preserved	O
in	O
XHM-ED	O
patients	O
because	O
APCs	B-cell_type
retain	O
the	O
capacity	O
to	O
respond	O
to	O
stimulation	O
by	O
lipopolysaccharide	O
or	O
Staphylococcus	B-protein
aureus	I-protein
Cowan	I-protein
's	I-protein
antigen	I-protein
(	O
SAC	B-protein
)	O
.	O

Overall	O
,	O
the	O
phenotype	O
observed	O
in	O
XHM-ED	O
patients	O
shows	O
that	O
the	O
putative	B-protein
zinc-finger	I-protein
domain	I-protein
of	O
NEMO	B-protein
has	O
a	O
regulatory	O
function	O
and	O
demonstrates	O
the	O
definite	O
requirement	O
of	O
CD40	B-protein
-mediated	O
NF-kappaB	O
activation	O
for	O
B	B-cell_type
cell	I-cell_type
immunoglobulin	O
class-switching	O
.	O

-DOCSTART-	O

Granulocytic	O
differentiation	O
of	O
human	B-cell_type
NB4	I-cell_type
promyelocytic	I-cell_type
leukemia	I-cell_type
cells	I-cell_type
induced	O
by	O
all-trans	O
retinoic	O
acid	O
metabolites	O
.	O

The	O
metabolism	O
of	O
all-trans	O
retinoic	O
acid	O
(	O
ATRA	O
)	O
has	O
been	O
reported	O
to	O
be	O
partly	O
responsible	O
for	O
the	O
in	O
vivo	O
resistance	O
to	O
ATRA	O
seen	O
in	O
the	O
treatment	O
of	O
human	O
acute	O
promyelocytic	O
leukemia	O
(	O
APL	O
)	O
.	O

However	O
,	O
ATRA	O
metabolism	O
appears	O
to	O
be	O
involved	O
in	O
the	O
growth	O
inhibition	O
of	O
several	O
cancer	B-cell_line
cell	I-cell_line
lines	I-cell_line
in	O
vitro	O
.	O

The	O
purpose	O
of	O
this	O
study	O
was	O
to	O
evaluate	O
the	O
in	O
vitro	O
activity	O
of	O
the	O
principal	O
metabolites	O
of	O
ATRA	O
[	O
4-hydroxy-retinoic	O
acid	O
(	O
4-OH-RA	O
)	O
,	O
18-hydroxy-retinoic	O
acid	O
(	O
18-OH-RA	O
)	O
,	O
4-oxo-retinoic	O
acid	O
(	O
4-oxo-RA	O
)	O
,	O
and	O
5	O
,	O
6-epoxy-retinoic	O
acid	O
(	O
5	O
,	O
6-epoxy-RA	O
)	O
]	O
in	O
NB4	B-cell_line
,	I-cell_line
a	I-cell_line
human	I-cell_line
promyelocytic	I-cell_line
leukemia	I-cell_line
cell	I-cell_line
line	I-cell_line
that	O
exhibits	O
the	O
APL	O
diagnostic	O
t	O
(	O
15	O
;	O
17	O
)	O
chromosomal	O
translocation	O
and	O
expresses	O
the	O
PML-RAR	B-protein
alpha	I-protein
fusion	I-protein
protein	I-protein
.	O

We	O
established	O
that	O
the	O
four	O
ATRA	O
metabolites	O
were	O
indeed	O
formed	O
by	O
the	O
NB4	B-cell_line
cells	I-cell_line
in	O
vitro	O
.	O

NB4	O
cell	O
growth	O
was	O
inhibited	O
(	O
69-78	O
%	O
at	O
120	O
h	O
)	O
and	O
cell	O
cycle	O
progression	O
in	O
the	O
G1	O
phase	O
(	O
82-85	O
%	O
at	O
120	O
h	O
)	O
was	O
blocked	O
by	O
ATRA	O
and	O
all	O
of	O
the	O
metabolites	O
at	O
1	O
microM	O
concentration	O
.	O

ATRA	O
and	O
its	O
metabolites	O
could	O
induce	O
NB4	B-cell_line
cells	I-cell_line
differentiation	O
with	O
similar	O
activity	O
,	O
as	O
evaluated	O
by	O
cell	O
morphology	O
,	O
by	O
the	O
nitroblue	O
tetrazolium	O
reduction	O
test	O
(	O
82-88	O
%	O
at	O
120	O
h	O
)	O
or	O
by	O
the	O
expression	O
of	O
the	O
maturation	B-protein
specific	I-protein
cell	I-protein
surface	I-protein
marker	I-protein
CD11c	I-protein
.	O

In	O
addition	O
,	O
nuclear	O
body	O
reorganization	O
to	O
macropunctated	O
structures	O
,	O
as	O
well	O
as	O
the	O
degradation	O
of	O
PML-RAR	B-protein
alpha	I-protein
,	O
was	O
found	O
to	O
be	O
similar	O
for	O
ATRA	O
and	O
all	O
of	O
its	O
metabolites	O
.	O

Comparison	O
of	O
the	O
relative	O
potency	O
of	O
the	O
retinoids	O
using	O
the	O
nitroblue	O
tetrazolium	O
reduction	O
test	O
showed	O
effective	O
concentrations	O
required	O
to	O
differentiate	O
50	O
%	O
of	O
cells	O
in	O
72	O
h	O
as	O
follows	O
:	O
ATRA	O
,	O
15.8	O
+/-	O
1.7	O
nM	O
;	O
4-oxo-RA	O
,	O
38.3	O
+/-	O
1.3	O
nM	O
;	O
18-OH-RA	O
,	O
55.5	O
+/-	O
1.8	O
nM	O
;	O
4-OH-RA	O
,	O
79.8	O
+/-	O
1.8	O
nM	O
;	O
and	O
5	O
,	O
6-epoxy-RA	O
,	O
99.5	O
+/-	O
1.5	O
nM	O
.	O

The	O
ATRA	O
metabolites	O
were	O
found	O
to	O
exert	O
their	O
differentiation	O
effects	O
via	O
the	O
RAR	B-protein
alpha	I-protein
nuclear	I-protein
receptors	I-protein
,	O
because	O
the	O
RAR	O
alpha-specific	O
antagonist	O
BMS614	O
blocked	O
metabolite-induced	O
CD11c	O
expression	O
in	O
NB4	B-cell_line
cells	I-cell_line
.	O

These	O
data	O
demonstrate	O
that	O
the	O
principal	O
ATRA	O
Phase	O
1	O
metabolites	O
can	O
elicit	O
leukemia	B-cell_type
cell	I-cell_type
growth	O
inhibition	O
and	O
differentiation	O
in	O
vitro	O
through	O
the	O
RAR	O
alpha	O
signaling	O
pathway	O
,	O
and	O
they	O
suggest	O
that	O
these	O
metabolites	O
may	O
play	O
a	O
role	O
in	O
ATRA	O
antileukemic	O
activity	O
in	O
vivo	O
.	O

-DOCSTART-	O

Expression	O
of	O
oestrogen	B-protein
and	I-protein
progesterone	I-protein
receptors	I-protein
by	O
mast	B-cell_type
cells	I-cell_type
alone	O
,	O
but	O
not	O
lymphocytes	B-cell_type
,	O
macrophages	B-cell_type
or	O
other	O
immune	B-cell_type
cells	I-cell_type
in	O
human	O
upper	O
airways	O
.	O

BACKGROUND	O
:	O
Nasal	O
polyposis	O
often	O
coexists	O
with	O
asthma	O
in	O
airway	O
inflammatory	O
conditions	O
characterised	O
by	O
the	O
infiltration	O
of	O
a	O
range	O
of	O
immune	B-cell_type
cells	I-cell_type
.	O

A	O
potentially	O
important	O
role	O
for	O
ovarian	O
hormones	O
has	O
been	O
implicated	O
in	O
airway	O
inflammation	O
but	O
the	O
cellular	O
target	O
for	O
such	O
action	O
is	O
not	O
known	O
.	O

METHODS	O
:	O
Expression	O
of	O
oestrogen	B-protein
receptors	I-protein
(	O
ER	B-protein
)	O
and	O
progesterone	B-protein
receptors	I-protein
(	O
PR	B-protein
)	O
was	O
examined	O
using	O
immunohistochemistry	O
in	O
formalin	O
fixed	O
nasal	O
polyp	O
tissues	O
from	O
47	O
subjects	O
.	O

The	O
cells	O
positive	O
for	O
ER	B-protein
or	O
PR	B-protein
were	O
confirmed	O
by	O
spatial	O
location	O
,	O
dual	O
immunolabelling	O
,	O
and	O
histochemical	O
staining	O
.	O

RESULTS	O
:	O
Consistent	O
with	O
the	O
known	O
features	O
of	O
nasal	O
polyps	O
,	O
CD4+	B-cell_type
(	I-cell_type
T	I-cell_type
helper/inducer	I-cell_type
)	I-cell_type
,	O
CD8+	B-cell_type
(	I-cell_type
cytotoxic/suppressor	I-cell_type
)	I-cell_type
,	O
CD68+	B-cell_type
(	O
macrophages	B-cell_type
)	O
,	O
mast	B-cell_type
cells	I-cell_type
,	O
eosinophils	B-cell_type
and	O
neutrophils	B-cell_type
were	O
all	O
clearly	O
detected	O
by	O
their	O
relevant	O
monoclonal	B-protein
antibodies	I-protein
or	O
appropriate	O
histochemical	O
staining	O
,	O
but	O
only	O
mast	B-cell_type
cells	I-cell_type
tested	O
positive	O
for	O
ER	B-protein
/PR	O
labelling	O
with	O
their	O
polyclonal	B-protein
and	I-protein
monoclonal	I-protein
antibodies	I-protein
.	O

The	O
frequencies	O
for	O
expression	O
were	O
61.7	O
%	O
for	O
ER	B-cell_type
positive	I-cell_type
and	O
59.6	O
%	O
for	O
PR	B-cell_type
positive	I-cell_type
cells	I-cell_type
.	O

The	O
expression	O
of	O
ER	B-protein
/	O
PR	B-protein
was	O
independent	O
of	O
patient	O
sex	O
and	O
age	O
but	O
was	O
highly	O
correlated	O
with	O
the	O
numbers	O
of	O
mast	B-cell_type
cells	I-cell_type
(	O
r	O
=	O
0.973	O
,	O
p	O
<	O
0.001	O
for	O
ER	B-protein
;	O
r	O
=	O
0.955	O
,	O
p	O
<	O
0.001	O
for	O
PR	B-protein
)	O
.	O

Fewer	O
than	O
5	O
%	O
of	O
mast	B-cell_type
cells	I-cell_type
were	O
found	O
to	O
be	O
negative	O
for	O
ER	B-protein
/	O
PR	B-protein
expression	O
.	O

CONCLUSIONS	O
:	O
Mast	B-cell_type
cells	I-cell_type
alone	O
,	O
but	O
not	O
lymphocytes	B-cell_type
,	O
macrophages	B-cell_type
,	O
or	O
other	O
immune	B-cell_type
cells	I-cell_type
,	O
express	O
ER	B-protein
/	O
PR	B-protein
in	O
human	O
upper	O
airways	O
.	O

Numerous	O
ER/PR	B-cell_type
positive	I-cell_type
mast	I-cell_type
cells	I-cell_type
exist	O
in	O
nasal	O
polyps	O
,	O
indicating	O
that	O
this	O
may	O
be	O
a	O
major	O
route	O
for	O
the	O
involvement	O
of	O
sex	O
hormones	O
in	O
airway	O
inflammation	O
when	O
exposed	O
to	O
the	O
higher	O
and	O
varying	O
concentration	O
of	O
oestrogen	O
and	O
progesterone	O
characteristic	O
of	O
females	O
.	O

-DOCSTART-	O

NF	O
kappa	O
b	O
signaling	O
in	O
posthypoxic	B-cell_type
endothelial	I-cell_type
cells	I-cell_type
:	O
relevance	O
to	O
E-selectin	O
expression	O
and	O
neutrophil	O
adhesion	O
.	O

Our	O
previous	O
studies	O
have	O
implicated	O
the	O
nuclear	O
transcription	O
factor	O
kappa	O
B	O
(	O
NF	B-protein
kappa	I-protein
B	I-protein
)	O
in	O
the	O
regulation	O
of	O
adhesion	B-protein
molecule	I-protein
expression	O
in	O
endothelial	B-cell_type
cells	I-cell_type
exposed	O
to	O
anoxia-reoxygenation	O
(	O
A/R	O
)	O
or	O
a	O
redox	O
imbalance	O
.	O

The	O
objectives	O
of	O
this	O
study	O
were	O
(	O
1	O
)	O
to	O
define	O
the	O
kinetics	O
of	O
NF	B-protein
kappa	I-protein
B	I-protein
activation	O
by	O
examining	O
I	O
kappa	O
B	O
alpha	O
degradation	O
and	O
the	O
nuclear	O
translocation	O
of	O
p65	B-protein
in	O
response	O
to	O
A/R	O
or	O
redox	O
imbalance	O
(	O
induced	O
by	O
treatment	O
of	O
cells	O
with	O
diamide	O
and	O
buthionine	O
sulfoximine	O
)	O
and	O
(	O
2	O
)	O
to	O
determine	O
whether	O
the	O
signal	O
for	O
I	O
kappa	O
B	O
alpha	O
degradation	O
,	O
nuclear	O
translocation	O
of	O
p65	B-protein
,	O
and	O
E-selectin	B-protein
-mediated	O
neutrophil	O
adhesion	O
is	O
related	O
to	O
the	O
activity	O
of	O
protein	B-protein
tyrosine	I-protein
kinase	I-protein
(	O
PTK	B-protein
)	O
,	O
protein	B-protein
tyrosine	I-protein
phosphatase	I-protein
(	O
PTPase	B-protein
)	O
and/or	O
protein	B-protein
kinase	I-protein
C	I-protein
(	O
PKC	B-protein
)	O
.	O

The	O
results	O
demonstrate	O
that	O
both	O
A/R	O
and	O
redox	O
imbalance	O
led	O
to	O
I	O
kappa	O
B	O
alpha	O
degradation	O
within	O
30	O
min	O
and	O
the	O
concomitant	O
appearance	O
of	O
p65	B-protein
in	O
the	O
nucleus	O
,	O
consistent	O
with	O
rapid	O
cytosolic	O
activation	O
of	O
NF	B-protein
kappa	I-protein
B	I-protein
and	O
subsequent	O
nuclear	O
translocation	O
of	O
the	O
activated	B-protein
p65	I-protein
subunit	I-protein
.	O

Inhibition	O
of	O
PKC	B-protein
blocked	O
I	B-protein
kappa	I-protein
B	I-protein
alpha	I-protein
degradation	O
and	O
p65	B-protein
translocation	O
in	O
A/R-challenged	O
,	O
but	O
not	O
redox-altered	O
,	O
endothelial	B-cell_type
cells	I-cell_type
.	O

However	O
,	O
both	O
A/R-	O
and	O
redox-induced	O
NF	O
kappa	O
B	O
activation	O
was	O
blocked	O
by	O
inhibition	O
of	O
PTK	B-protein
.	O

Similarly	O
,	O
A/R-induced	O
E-selectin	O
expression	O
and	O
neutrophil-endothelial	O
cell	O
adhesion	O
were	O
blocked	O
by	O
inhibition	O
of	O
PKC	B-protein
or	O
PTK	B-protein
,	O
while	O
only	O
PTK	B-protein
inhibited	O
the	O
redox-induced	O
adhesion	O
response	O
.	O

Pretreatment	O
of	O
cells	O
with	O
N-acetyl	O
cysteine	O
effectively	O
blocked	O
A/R-	O
or	O
redox-induced	O
I	O
kappa	O
B	O
degradation	O
and	O
significantly	O
attenuated	O
the	O
respective	O
neutrophil	O
adhesion	O
responses	O
.	O

Collectively	O
,	O
these	O
findings	O
indicate	O
that	O
A/R-induced	O
E-selectin	O
expression	O
and	O
neutrophil-endothelial	O
cell	O
adhesion	O
are	O
mediated	O
by	O
both	O
PKC	B-protein
and	O
PTK	B-protein
,	O
which	O
signal	O
rapid	O
activation	O
of	O
NF	B-protein
kappa	I-protein
B	I-protein
.	O

This	O
A/R-induced	O
NF	B-protein
kappa	I-protein
B	I-protein
signaling	O
response	O
appears	O
to	O
be	O
mediated	O
,	O
at	O
least	O
in	O
part	O
,	O
by	O
intracellular	O
redox	O
imbalance	O
.	O

Copyright	O
2001	O
S.	O
Karger	O
AG	O
,	O
Basel	O

-DOCSTART-	O

Induction	O
of	O
apoptosis	O
in	O
human	B-cell_type
lymphocytes	I-cell_type
by	O
the	O
herbicide	O
2	O
,	O
4-dichlorophenoxyacetic	O
acid	O
.	O

Dimethylammonium	O
salt	O
of	O
2	O
,	O
4-dichlorophenoxyacetic	O
acid	O
(	O
DMA-2	O
,	O
4-D	O
)	O
is	O
a	O
widely	O
used	O
herbicide	O
that	O
is	O
considered	O
moderately	O
toxic	O
.	O

In	O
the	O
present	O
study	O
we	O
found	O
that	O
DMA-2	O
,	O
4-D	O
is	O
able	O
to	O
cause	O
apoptosis	O
in	O
peripheral	B-cell_type
blood	I-cell_type
lymphocytes	I-cell_type
of	O
healthy	O
individuals	O
and	O
Jurkat	B-cell_line
T	I-cell_line
cells	I-cell_line
.	O

Apoptosis	O
induced	O
by	O
DMA-2	O
,	O
4-D	O
was	O
dose	O
and	O
time	O
dependent	O
,	O
independent	O
of	O
Fas	B-protein
,	O
TNF	B-protein
receptor	I-protein
1	I-protein
or	O
the	O
aromatic	B-protein
hydrocarbon	I-protein
receptor	I-protein
,	O
and	O
involved	O
disruption	O
of	O
the	O
mitochondrial	O
transmembrane	O
potential	O
and	O
activation	O
of	O
caspase-9	B-protein
.	O

ZVAD-FMK	O
,	O
a	O
broad-spectrum	O
inhibitor	O
of	O
caspases	B-protein
,	O
blocked	O
DMA-2	O
,	O
4-D-induced	O
apoptosis	O
completely	O
.	O

While	O
an	O
inhibitor	O
of	O
caspase-9	B-protein
,	O
as	O
well	O
as	O
caspase-9	B-protein
and	O
caspase-3	O
inhibitors	O
in	O
combination	O
,	O
strongly	O
blocked	O
DMA-2	O
,	O
4-D-induced	O
apoptosis	O
,	O
an	O
inhibitor	O
of	O
caspase-3	B-protein
had	O
a	O
moderate	O
inhibitory	O
effect	O
.	O

Unlike	O
Fas	B-protein
-mediated	O
apoptosis	O
,	O
the	O
initiator	B-protein
caspase	I-protein
,	O
caspase-8	B-protein
,	O
was	O
not	O
involved	O
in	O
DMA-2	O
,	O
4-D-induced	O
apoptosis	O
.	O

Transfection	O
of	O
Jurkat	B-cell_line
cells	I-cell_line
with	O
Bcl-2	B-protein
prevented	O
DMA-2	O
,	O
4-D-induced	O
disruption	O
of	O
the	O
mitochondrial	O
transmembrane	O
potential	O
and	O
led	O
to	O
a	O
complete	O
blockage	O
of	O
apoptosis	O
.	O

Our	O
data	O
indicate	O
that	O
DMA-2	O
,	O
4-D	O
kills	O
human	B-cell_type
lymphocytes	I-cell_type
by	O
initiating	O
apoptosis	O
via	O
a	O
direct	O
effect	O
on	O
mitochondria	O
.	O

The	O
activation	O
of	O
caspases	B-protein
occurs	O
downstream	O
of	O
mitochondrial	O
damage	O
,	O
and	O
the	O
dysfunction	O
of	O
mitochondria	O
appears	O
to	O
be	O
sufficient	O
for	O
triggering	O
all	O
downstream	O
events	O
leading	O
to	O
apoptosis	O
.	O

-DOCSTART-	O

Inhibition	O
of	O
Th1	O
differentiation	O
by	O
IL-6	B-protein
is	O
mediated	O
by	O
SOCS1	B-protein
.	O

Interleukin	B-protein
6	I-protein
(	O
IL-6	B-protein
)	O
is	O
a	O
cytokine	B-protein
produced	O
by	O
immune	B-cell_type
and	I-cell_type
nonimmune	I-cell_type
cells	I-cell_type
and	O
exhibits	O
functional	O
pleiotropy	O
and	O
redundancy	O
.	O

IL-6	B-protein
plays	O
an	O
important	O
role	O
in	O
the	O
differentiation	O
of	O
several	O
cell	O
types	O
.	O

Here	O
,	O
we	O
describe	O
a	O
novel	O
function	O
of	O
IL-6	B-protein
:	O
the	O
negative	O
regulation	O
of	O
CD4	B-protein
+	O
Th1	O
cell	O
differentiation	O
.	O

While	O
IL-6	B-protein
-directed	O
CD4	B-protein
+	O
Th2	O
differentiation	O
is	O
mediated	O
by	O
IL-4	B-protein
,	O
inhibition	O
of	O
Th1	O
differentiation	O
by	O
IL-6	B-protein
is	O
independent	O
of	O
IL-4	B-protein
.	O

IL-6	B-protein
upregulates	O
suppressor	B-protein
of	I-protein
cytokine	I-protein
signaling	I-protein
1	I-protein
(	O
SOCS1	B-protein
)	O
expression	O
in	O
activated	B-cell_type
CD4+	I-cell_type
T	I-cell_type
cells	I-cell_type
,	O
thereby	O
interfering	O
with	O
signal	B-protein
transducer	I-protein
and	I-protein
activator	I-protein
of	I-protein
transcription	I-protein
1	I-protein
(	O
STAT1	B-protein
)	O
phosphorylation	O
induced	O
by	O
interferon	B-protein
gamma	I-protein
(	O
IFNgamma	B-protein
)	O
.	O

Inhibition	O
of	O
IFNgamma	B-protein
receptor-mediated	O
signals	O
by	O
IL-6	B-protein
prevents	O
autoregulation	O
of	O
IFNgamma	B-DNA
gene	I-DNA
expression	O
by	O
IFNgamma	B-protein
during	O
CD4+	B-cell_type
T	I-cell_type
cell	I-cell_type
activation	O
,	O
thereby	O
preventing	O
Th1	O
differentiation	O
.	O

Thus	O
,	O
IL-6	B-protein
promotes	O
CD4	B-protein
+	O
Th2	O
differentiation	O
and	O
inhibits	O
Th1	O
differentiation	O
by	O
two	O
independent	O
molecular	O
mechanisms	O
.	O

-DOCSTART-	O

Interaction	O
between	O
CCAAT/enhancer	B-protein
binding	I-protein
protein	I-protein
and	O
cyclic	B-protein
AMP	I-protein
response	I-protein
element	I-protein
binding	I-protein
protein	I-protein
1	I-protein
regulates	O
human	O
immunodeficiency	O
virus	O
type	O
1	O
transcription	O
in	O
cells	O
of	O
the	O
monocyte/macrophage	B-cell_type
lineage	I-cell_type
.	O

Recent	O
observations	O
have	O
shown	O
two	O
CCAAT/enhancer	B-DNA
binding	I-DNA
protein	I-DNA
(	I-DNA
C/EBP	I-DNA
)	I-DNA
binding	I-DNA
sites	I-DNA
to	O
be	O
critically	O
important	O
for	O
efficient	O
human	O
immunodeficiency	O
virus	O
type	O
1	O
(	O
HIV-1	O
)	O
replication	O
within	O
cells	O
of	O
the	O
monocyte/macrophage	B-cell_type
lineage	I-cell_type
,	O
a	O
cell	O
type	O
likely	O
involved	O
in	O
transport	O
of	O
the	O
virus	O
to	O
the	O
brain	O
.	O

Additionally	O
,	O
sequence	O
variation	O
at	O
C/EBP	B-DNA
site	I-DNA
I	I-DNA
,	O
which	O
lies	O
immediately	O
upstream	O
of	O
the	O
distal	B-DNA
nuclear	I-DNA
factor	I-DNA
kappa	I-DNA
B	I-DNA
site	I-DNA
and	O
immediately	O
downstream	O
of	O
a	O
binding	O
site	O
for	O
activating	B-protein
transcription	I-protein
factor	I-protein
(	O
ATF	B-protein
)	O
/	O
cyclic	B-protein
AMP	I-protein
response	I-protein
element	I-protein
binding	I-protein
protein	I-protein
(	O
CREB	B-protein
)	O
,	O
has	O
been	O
shown	O
to	O
affect	O
HIV-1	B-DNA
long	I-DNA
terminal	I-DNA
repeat	I-DNA
(	O
LTR	B-DNA
)	O
activity	O
.	O

Given	O
that	O
C/EBP	B-protein
proteins	I-protein
have	O
been	O
shown	O
to	O
interact	O
with	O
many	O
other	O
transcription	B-protein
factors	I-protein
including	O
members	O
of	O
the	O
ATF/CREB	B-protein
family	I-protein
,	O
we	O
proceeded	O
to	O
determine	O
whether	O
an	O
adjacent	O
ATF/CREB	B-DNA
binding	I-DNA
site	I-DNA
could	O
affect	O
C/EBP	B-protein
protein	I-protein
binding	O
to	O
C/EBP	B-DNA
site	I-DNA
I	I-DNA
.	O

Electrophoretic	O
mobility	O
shift	O
analyses	O
indicated	O
that	O
selected	O
ATF/CREB	B-DNA
site	I-DNA
variants	I-DNA
assisted	O
in	O
the	O
recruitment	O
of	O
C/EBP	B-protein
proteins	I-protein
to	O
an	O
adjacent	O
,	O
naturally	O
occurring	O
,	O
low-affinity	O
C/EBP	B-DNA
site	I-DNA
.	O

This	O
biophysical	O
interaction	O
appears	O
to	O
occur	O
via	O
at	O
least	O
two	O
mechanisms	O
.	O

First	O
,	O
low	O
amounts	O
of	O
CREB-1	B-protein
and	O
C/EBP	B-protein
appear	O
to	O
heterodimerize	O
and	O
bind	O
to	O
a	O
site	O
consisting	O
of	O
a	O
half	O
site	O
from	O
both	O
the	O
ATF/CREB	B-DNA
and	I-DNA
C/EBP	I-DNA
binding	I-DNA
sites	I-DNA
.	O

In	O
addition	O
,	O
CREB-1	B-protein
homodimers	I-protein
bind	O
to	O
the	O
ATF/CREB	B-DNA
site	I-DNA
and	O
recruit	O
C/EBP	B-protein
dimers	I-protein
to	O
their	O
cognate	O
weak	B-DNA
binding	I-DNA
sites	I-DNA
.	O

This	O
interaction	O
is	O
reciprocal	O
,	O
since	O
C/EBP	B-protein
dimer	I-protein
binding	O
to	O
a	O
strong	O
C/EBP	B-DNA
site	I-DNA
leads	O
to	O
enhanced	O
CREB-1	B-protein
recruitment	O
to	O
ATF/CREB	B-DNA
sites	I-DNA
that	O
are	O
weakly	O
bound	O
by	O
CREB	B-protein
.	O

Sequence	O
variation	O
at	O
both	O
C/EBP	B-DNA
and	I-DNA
ATF/CREB	I-DNA
sites	I-DNA
affects	O
the	O
molecular	O
interactions	O
involved	O
in	O
mediating	O
both	O
of	O
these	O
mechanisms	O
.	O

Most	O
importantly	O
,	O
sequence	O
variation	O
at	O
the	O
ATF/CREB	B-DNA
binding	I-DNA
site	I-DNA
affected	O
basal	O
LTR	B-DNA
activity	O
as	O
well	O
as	O
LTR	B-DNA
function	O
following	O
interleukin-6	B-protein
stimulation	O
,	O
a	O
treatment	O
that	O
leads	O
to	O
increases	O
in	O
C/EBP	O
activation	O
.	O

Thus	O
,	O
HIV-1	B-DNA
LTR	I-DNA
ATF/CREB	B-DNA
binding	I-DNA
site	I-DNA
sequence	O
variation	O
may	O
modulate	O
cellular	O
signaling	O
at	O
the	O
viral	B-DNA
promoter	I-DNA
through	O
the	O
C/EBP	O
pathway	O

-DOCSTART-	O

Selective	O
inhibition	O
of	O
interleukin-4	O
gene	O
expression	O
in	O
human	B-cell_type
T	I-cell_type
cells	I-cell_type
by	O
aspirin	O
.	O

Previous	O
studies	O
indicated	O
that	O
aspirin	O
(	O
acetylsalicylic	O
acid	O
[	O
ASA	O
]	O
)	O
can	O
have	O
profound	O
immunomodulatory	O
effects	O
by	O
regulating	O
cytokine	O
gene	O
expression	O
in	O
several	O
types	O
of	O
cells	O
.	O

This	O
study	O
is	O
the	O
first	O
in	O
which	O
concentrations	O
of	O
ASA	O
in	O
the	O
therapeutic	O
range	O
were	O
found	O
to	O
significantly	O
reduce	O
interleukin	B-protein
(	I-protein
IL	I-protein
)	I-protein
-4	I-protein
secretion	O
and	O
RNA	O
expression	O
in	O
freshly	B-cell_type
isolated	I-cell_type
and	I-cell_type
mitogen-primed	I-cell_type
human	I-cell_type
CD4+	I-cell_type
T	I-cell_type
cells	I-cell_type
.	O

In	O
contrast	O
,	O
ASA	O
did	O
not	O
affect	O
IL-13	B-protein
,	O
interferon-gamma	B-protein
,	O
and	O
IL-2	B-protein
expression	O
.	O

ASA	O
inhibited	O
IL-4	B-protein
,	O
but	O
not	O
IL-2	B-protein
,	O
promoter-driven	O
chloramphenicol	O
acetyltransferase	O
expression	O
in	O
transiently	O
transfected	O
Jurkat	B-cell_line
T	I-cell_line
cells	I-cell_line
.	O

The	O
structurally	O
unrelated	O
nonsteroidal	O
anti-inflammatory	O
drugs	O
indomethacin	O
and	O
flurbiprofen	O
did	O
not	O
affect	O
cytokine	O
gene	O
expression	O
in	O
T	B-cell_type
cells	I-cell_type
,	O
whereas	O
the	O
weak	O
cyclo-oxygenase	O
inhibitor	O
salicylic	O
acid	O
was	O
at	O
least	O
as	O
effective	O
as	O
ASA	O
in	O
inhibiting	O
IL-4	B-protein
expression	O
and	O
promoter	O
activity	O
.	O

The	O
inhibitory	O
effect	O
of	O
ASA	O
on	O
IL-4	O
transcription	O
was	O
not	O
mediated	O
by	O
decreased	O
nuclear	O
expression	O
of	O
the	O
known	O
salicylate	B-protein
target	I-protein
nuclear	I-protein
factor	I-protein
(	I-protein
NF	I-protein
)	I-protein
-kappaB	I-protein
and	O
was	O
accompanied	O
by	O
reduced	O
binding	O
of	O
an	O
inducible	O
factor	O
to	O
an	O
IL-4	B-DNA
promoter	I-DNA
region	I-DNA
upstream	O
of	O
,	O
but	O
not	O
overlapping	O
,	O
the	O
NF	B-DNA
of	I-DNA
activated	I-DNA
T	I-DNA
cells-	I-DNA
and	I-DNA
NF-kappaB-binding	I-DNA
P1	I-DNA
element	I-DNA
.	O

It	O
is	O
concluded	O
that	O
anti-inflammatory	O
salicylates	O
,	O
by	O
means	O
of	O
a	O
previously	O
unrecognized	O
mechanism	O
of	O
action	O
,	O
can	O
influence	O
the	O
nature	O
of	O
adaptive	O
immune	O
responses	O
by	O
selectively	O
inhibiting	O
the	O
expression	O
of	O
IL-4	B-protein
,	O
a	O
critical	O
effector	O
of	O
these	O
responses	O
,	O
in	O
CD4+	B-cell_type
T	I-cell_type
cells	I-cell_type
.	O

-DOCSTART-	O

Molecular	O
and	O
cellular	O
mediators	O
of	O
interleukin-1	B-protein
-dependent	O
acute	O
inflammatory	O
arthritis	O
.	O

OBJECTIVE	O
:	O
To	O
examine	O
the	O
molecular	O
and	O
cellular	O
mechanisms	O
in	O
a	O
model	O
of	O
acute	O
inflammatory	O
monarticular	O
arthritis	O
induced	O
by	O
methylated	B-protein
bovine	I-protein
serum	I-protein
albumin	I-protein
(	O
mBSA	B-protein
)	O
and	O
interleukin-1	B-protein
(	O
IL-1	B-protein
)	O
.	O

METHODS	O
:	O
Mice	O
were	O
injected	O
intraarticularly	O
with	O
mBSA	B-protein
on	O
day	O
0	O
and	O
subcutaneously	O
with	O
recombinant	B-protein
human	I-protein
IL-1beta	I-protein
on	O
days	O
0-2	O
.	O

At	O
day	O
7	O
,	O
knee	O
joints	O
were	O
removed	O
and	O
assessed	O
histologically	O
.	O

Flow	O
cytometry	O
and	O
RNase	O
protection	O
were	O
used	O
to	O
analyze	O
IL-1	B-protein
-dependent	O
events	O
.	O

RESULTS	O
:	O
C57BL/6	O
(	O
B6	O
)	O
,	O
129/Sv	O
,	O
and	O
(	O
B6	O
x	O
129/	O
Sv	O
)	O
F1	O
hybrid	O
mice	O
,	O
all	O
H-2b	O
strains	O
,	O
were	O
susceptible	O
to	O
mBSA	B-protein
/IL-1-induced	O
arthritis	O
,	O
whereas	O
C3H/HeJ	O
(	O
H-2k	O
)	O
mice	O
were	O
not	O
.	O

B6	O
mice	O
lacking	O
T	B-cell_type
and	I-cell_type
B	I-cell_type
cells	I-cell_type
(	O
RAG1-/-	O
)	O
or	O
major	B-protein
histocompatibility	I-protein
complex	I-protein
(	I-protein
MHC	I-protein
)	I-protein
class	I-protein
II	I-protein
antigens	I-protein
(	O
MHCII-/-	O
)	O
,	O
and	O
B6	O
mice	O
treated	O
with	O
a	O
CD4+	B-protein
T	I-protein
cell-depleting	I-protein
monoclonal	I-protein
antibody	I-protein
,	O
were	O
resistant	O
to	O
disease	O
.	O

In	O
contrast	O
,	O
B	O
cell-deficient	O
(	O
muMT/	O
muMT	O
)	O
mice	O
developed	O
arthritis	O
at	O
an	O
incidence	O
and	O
severity	O
similar	O
to	O
that	O
of	O
controls	O
.	O

RelB-deficient	O
(	O
RelB-/-	O
)	O
bone	O
marrow	O
chimeric	O
mice	O
had	O
arthritis	O
that	O
was	O
significantly	O
reduced	O
in	O
incidence	O
and	O
severity	O
.	O

In	O
B6	O
mice	O
,	O
flow	O
cytometry	O
demonstrated	O
an	O
IL-1-dependent	B-cell_type
leukocyte	I-cell_type
infiltration	O
into	O
the	O
synovial	O
compartment	O
and	O
RNase	O
protection	O
assays	O
revealed	O
induction	O
of	O
messenger	B-RNA
RNA	I-RNA
(	O
mRNA	B-RNA
)	O
for	O
the	O
chemokines	B-protein
monocyte	B-protein
chemoattractant	I-protein
protein	I-protein
1	I-protein
,	O
macrophage	B-protein
inhibitory	I-protein
protein	I-protein
2	I-protein
(	O
MIP-2	B-protein
)	O
,	O
RANTES	B-protein
,	O
MIP-1alpha	B-protein
,	O
and	O
MIP-1beta	B-protein
,	O
in	O
vivo	O
and	O
in	O
vitro	O
.	O

CONCLUSION	O
:	O
Arthritis	O
induced	O
by	O
mBSA	B-protein
/	O
IL-1	B-protein
is	O
strain	O
specific	O
and	O
dependent	O
on	O
CD4+	B-cell_type
T	I-cell_type
lymphocytes	I-cell_type
and	O
at	O
least	O
partially	O
on	O
RelB	B-protein
,	O
but	O
not	O
on	O
B	B-cell_type
lymphocytes	I-cell_type
or	O
antibody	B-protein
.	O

IL-1	B-protein
contributes	O
to	O
leukocyte	O
recruitment	O
to	O
the	O
synovium	O
and	O
directly	O
induces	O
chemokine	O
mRNA	O
production	O
by	O
synovial	B-cell_type
cells	I-cell_type
.	O

This	O
model	O
of	O
acute	O
monarticular	O
arthritis	O
is	O
particularly	O
suitable	O
for	O
further	O
investigations	O
into	O
cell-mediated	O
immunity	O
in	O
arthritis	O
and	O
the	O
role	O
of	O
IL-1	B-protein
.	O

-DOCSTART-	O

Positive	O
and	O
negative	O
regulation	O
of	O
granulopoiesis	O
by	O
endogenous	B-protein
RARalpha	I-protein
.	O

Acute	O
promyelocytic	O
leukemia	O
(	O
APL	O
)	O
is	O
always	O
associated	O
with	O
chromosomal	O
translocations	O
that	O
disrupt	O
the	O
retinoic	B-DNA
acid	I-DNA
receptor	I-DNA
alpha	I-DNA
(	I-DNA
RARalpha	I-DNA
)	I-DNA
gene	I-DNA
.	O

Whether	O
these	O
translocations	O
relate	O
to	O
a	O
role	O
for	O
endogenous	B-protein
RARalpha	I-protein
in	O
normal	O
granulopoiesis	O
remains	O
uncertain	O
because	O
most	O
studies	O
addressing	O
this	O
question	O
have	O
used	O
non-physiological	O
overexpression	O
systems	O
.	O

Granulocyte	O
differentiation	O
in	O
cells	O
derived	O
from	O
RARalpha-deficient	O
(	O
RARalpha	O
(	O
-/-	O
)	O
)	O
mice	O
was	O
studied	O
and	O
evaluated	O
in	O
the	O
context	O
of	O
agonist-bound	B-protein
and	I-protein
ligand-free	I-protein
RARalpha	I-protein
.	O

Our	O
results	O
demonstrate	O
that	O
RARalpha	B-protein
is	O
dispensable	O
for	O
granulopoiesis	O
,	O
as	O
RARalpha	O
(	O
-/-	O
)	O
mice	O
have	O
a	O
normal	B-cell_type
granulocyte	I-cell_type
population	I-cell_type
despite	O
an	O
impaired	O
ability	O
to	O
respond	O
to	O
retinoids	O
.	O

However	O
,	O
although	O
it	O
is	O
not	O
absolutely	O
required	O
,	O
RARalpha	B-protein
can	O
bidirectionally	O
modulate	O
granulopoiesis	O
.	O

RARalpha	B-protein
stimulates	O
differentiation	O
in	O
response	O
to	O
exogenous	O
retinoic	O
acid	O
.	O

Furthermore	O
,	O
endogenous	O
retinoids	O
control	O
granulopoiesis	O
in	O
vivo	O
,	O
as	O
either	O
vitamin	O
A-deficient	O
mice	O
or	O
animals	O
treated	O
with	O
an	O
RAR	O
antagonist	O
accumulate	O
more	O
immature	B-cell_type
granulocytes	I-cell_type
in	O
their	O
bone	O
marrow	O
.	O

Conversely	O
,	O
RARalpha	B-protein
acts	O
to	O
limit	O
differentiation	O
in	O
the	O
absence	O
of	O
ligand	B-protein
because	O
granulocyte	B-cell_type
precursors	I-cell_type
from	O
RARalpha	O
(	O
-/-	O
)	O
mice	O
differentiate	O
earlier	O
in	O
culture	O
.	O

Thus	O
,	O
the	O
block	O
in	O
granulopoiesis	O
exerted	O
by	O
RARalpha	B-protein
fusion	B-protein
proteins	I-protein
expressed	O
in	O
APL	B-cell_type
cells	I-cell_type
may	O
correspond	O
to	O
an	O
amplification	O
of	O
a	O
normal	O
function	O
of	O
unliganded	B-protein
RARalpha	I-protein
.	O

-DOCSTART-	O

Expression	O
and	O
function	O
of	O
a	O
stem	B-DNA
cell	I-DNA
promoter	I-DNA
for	O
the	O
murine	B-DNA
CBFalpha2	I-DNA
gene	I-DNA
:	O
distinct	O
roles	O
and	O
regulation	O
in	O
natural	B-cell_type
killer	I-cell_type
and	I-cell_type
T	I-cell_type
cell	I-cell_type
development	O
.	O

The	O
Runt	B-protein
family	I-protein
transcription	I-protein
factor	I-protein
CBFalpha2	B-protein
(	O
AML1	B-protein
,	O
PEBP2alphaB	B-protein
,	O
or	O
Runx1	B-protein
)	O
is	O
required	O
by	O
hematopoietic	B-cell_type
stem	I-cell_type
cells	I-cell_type
and	O
expressed	O
at	O
high	O
levels	O
in	O
T-lineage	B-cell_type
cells	I-cell_type
.	O

In	O
human	B-cell_type
T	I-cell_type
cells	I-cell_type
CBFalpha2	B-protein
is	O
usually	O
transcribed	O
from	O
a	O
different	B-DNA
promoter	I-DNA
(	O
distal	B-DNA
promoter	I-DNA
)	O
than	O
in	O
myeloid	B-cell_type
cells	I-cell_type
(	O
proximal	B-DNA
promoter	I-DNA
)	O
,	O
but	O
the	O
developmental	O
and	O
functional	O
significance	O
of	O
this	O
promoter	O
switch	O
has	O
not	O
been	O
known	O
.	O

Here	O
,	O
we	O
report	O
that	O
both	O
coding	B-DNA
and	I-DNA
noncoding	I-DNA
sequences	I-DNA
of	O
the	O
distal	B-DNA
5	I-DNA
'	I-DNA
end	I-DNA
are	O
highly	O
conserved	O
between	O
the	O
human	B-DNA
and	I-DNA
the	I-DNA
murine	I-DNA
genes	I-DNA
,	O
and	O
the	O
distal	B-DNA
promoter	I-DNA
is	O
responsible	O
for	O
the	O
overwhelming	O
majority	O
of	O
CBFalpha2	O
expression	O
in	O
murine	O
hematopoietic	B-cell_type
stem	I-cell_type
cells	I-cell_type
as	O
well	O
as	O
in	O
T	B-cell_type
cells	I-cell_type
.	O

Distal	O
promoter	O
activity	O
is	O
maintained	O
throughout	O
T	O
cell	O
development	O
and	O
at	O
lower	O
levels	O
in	O
B	O
cell	O
development	O
,	O
but	O
downregulated	O
in	O
natural	O
killer	O
cell	O
development	O
.	O

The	O
distal	B-DNA
N-terminal	I-DNA
isoform	I-DNA
binds	O
to	O
functionally	O
important	O
regulatory	B-DNA
sites	I-DNA
from	O
known	O
target	B-DNA
genes	I-DNA
with	O
two-	O
to	O
threefold	O
higher	O
affinity	O
than	O
the	O
proximal	B-DNA
N-terminal	I-DNA
isoform	I-DNA
.	O

Neither	O
full-length	B-DNA
isoform	I-DNA
alters	O
growth	O
of	O
a	O
myeloid	B-cell_line
cell	I-cell_line
line	I-cell_line
under	O
nondifferentiating	O
conditions	O
,	O
but	O
the	O
proximal	B-DNA
isoform	I-DNA
selectively	O
delays	O
mitotic	O
arrest	O
of	O
the	O
cell	O
line	O
under	O
differentiating	O
conditions	O
,	O
resulting	O
in	O
the	O
generation	O
of	O
greater	O
numbers	O
of	O
neutrophils	B-cell_type
.	O

-DOCSTART-	O

Constitutive	O
PI3-K	O
activity	O
is	O
essential	O
for	O
proliferation	O
,	O
but	O
not	O
survival	O
,	O
of	O
Theileria	B-cell_type
parva-transformed	I-cell_type
B	I-cell_type
cells	I-cell_type
.	O

Theileria	O
is	O
an	O
intracellular	O
parasite	O
that	O
causes	O
lymphoproliferative	O
disorders	O
in	O
cattle	O
,	O
and	O
infection	O
of	O
leucocytes	B-cell_type
induces	O
a	O
transformed	O
phenotype	O
similar	O
to	O
tumour	B-cell_type
cells	I-cell_type
,	O
but	O
the	O
mechanisms	O
by	O
which	O
the	O
parasite	O
induces	O
this	O
phenotype	O
are	O
not	O
understood	O
.	O

Here	O
,	O
we	O
show	O
that	O
infected	B-cell_type
B	I-cell_type
lymphocytes	I-cell_type
display	O
constitutive	O
phosphoinositide	O
3-kinase	O
(	O
PI3-K	O
)	O
activity	O
,	O
which	O
appears	O
to	O
be	O
necessary	O
for	O
proliferation	O
,	O
but	O
not	O
survival	O
.	O

Importantly	O
,	O
we	O
demonstrate	O
that	O
one	O
mechanism	O
by	O
which	O
PI3-K	B-protein
mediates	O
the	O
proliferation	O
of	O
infected	B-cell_type
B	I-cell_type
lymphocytes	I-cell_type
is	O
through	O
the	O
induction	O
of	O
a	O
granulocyte-monocyte	B-protein
colony-stimulating	I-protein
factor	I-protein
(	O
GM-CSF	B-protein
)	O
-dependent	O
autocrine	O
loop	O
.	O

PI3-K	B-protein
induction	O
of	O
GM-CSF	B-protein
appears	O
to	O
be	O
at	O
the	O
transcriptional	O
level	O
and	O
,	O
consistently	O
,	O
we	O
demonstrate	O
that	O
PI3-K	B-protein
is	O
also	O
involved	O
in	O
the	O
constitutive	O
induction	O
of	O
AP-1	B-protein
and	O
NF-kappaB	B-protein
,	O
which	O
characterizes	O
Theileria-infected	O
leucocytes	B-cell_type
.	O

Taken	O
together	O
,	O
our	O
results	O
highlight	O
a	O
novel	O
strategy	O
exploited	O
by	O
the	O
intracellular	O
parasite	O
Theileria	O
to	O
induce	O
continued	O
proliferation	O
of	O
its	O
host	B-cell_type
leucocyte	I-cell_type
.	O

-DOCSTART-	O

Lymphokine	B-protein
dependence	O
of	O
STAT3	O
activation	O
produced	O
by	O
surface	O
immunoglobulin	O
cross-linking	O
and	O
by	O
phorbol	O
ester	O
plus	O
calcium	O
ionophore	O
treatment	O
in	O
B	B-cell_type
cells	I-cell_type
.	O

Stimulation	O
of	O
B	B-cell_type
cells	I-cell_type
by	O
surface	B-protein
immunoglobulin	I-protein
(	O
sIg	B-protein
)	O
triggering	O
,	O
or	O
through	O
the	O
mitogenic	O
combination	O
of	O
phorbol	O
ester	O
and	O
calcium	O
ionophore	O
,	O
is	O
accompanied	O
by	O
activation	O
of	O
STAT	B-protein
transcription	I-protein
factors	I-protein
.	O

The	O
mechanism	O
responsible	O
for	O
the	O
delayed	O
nuclear	O
accumulation	O
of	O
phosphorylated	B-protein
STAT3	I-protein
was	O
examined	O
in	O
detail	O
,	O
focusing	O
on	O
the	O
role	O
of	O
B	B-protein
cell-derived	I-protein
lymphokines	I-protein
.	O

sIg	B-protein
-induced	O
activation	O
of	O
STAT3	B-protein
was	O
partially	O
inhibited	O
in	O
B	B-cell_type
cells	I-cell_type
obtained	O
from	O
IL-6-	O
or	O
IL-10-deficient	O
mice	O
,	O
and	O
was	O
partially	O
blocked	O
by	O
neutralizing	B-protein
antibodies	I-protein
directed	O
against	O
either	O
of	O
these	O
lymphokines	B-protein
.	O

sIg	B-protein
-induced	O
STAT3	O
activation	O
was	O
completely	O
inhibited	O
by	O
combining	O
IL-6-	B-protein
and	I-protein
IL-10-specific	I-protein
neutralizing	I-protein
antibodies	I-protein
,	O
or	O
by	O
adding	O
individual	O
neutralizing	B-protein
antibodies	I-protein
to	O
B	B-cell_type
cells	I-cell_type
obtained	O
from	O
lymphokine-deficient	O
animals	O
.	O

In	O
contrast	O
,	O
IL-10	B-protein
alone	O
appeared	O
to	O
account	O
for	O
STAT3	O
activation	O
resulting	O
from	O
B	O
cell	O
stimulation	O
with	O
phorbol	O
ester	O
and	O
calcium	O
ionophore	O
.	O

In	O
keeping	O
with	O
these	O
results	O
,	O
soluble	O
IL-6	B-protein
and	O
IL-10	B-protein
were	O
found	O
in	O
supernatant	O
fluid	O
obtained	O
from	O
stimulated	B-cell_type
B	I-cell_type
cells	I-cell_type
.	O

This	O
work	O
indicates	O
that	O
a	O
lymphokine	O
pathway	O
is	O
responsible	O
for	O
STAT3	O
activation	O
that	O
occurs	O
late	O
after	O
B	O
cell	O
stimulation	O
,	O
and	O
points	O
out	O
differences	O
in	O
B	O
cell	O
activation	O
that	O
result	O
from	O
stimulation	O
through	O
the	O
antigen	B-protein
receptor	I-protein
and	O
through	O
pharmacological	O
mimicry	O
of	O
signaling	B-protein
mediators	I-protein
.	O

-DOCSTART-	O

Transcription	B-protein
factor	I-protein
AP-4	I-protein
is	O
a	O
ligand	B-protein
for	O
immunoglobulin-kappa	B-DNA
promoter	I-DNA
E-box	I-DNA
elements	I-DNA
.	O

Immunoglobulin	B-DNA
(	I-DNA
Ig	I-DNA
)	I-DNA
-kappa	I-DNA
promoters	I-DNA
from	O
humans	O
and	O
mice	O
share	O
conserved	B-DNA
sequences	I-DNA
.	O

The	O
octamer	B-DNA
element	I-DNA
is	O
common	O
to	O
all	O
Ig	B-DNA
promoters	I-DNA
and	O
pivotal	O
for	O
their	O
function	O
.	O

However	O
,	O
other	O
conserved	B-DNA
sequence	I-DNA
motifs	I-DNA
,	O
that	O
differ	O
between	O
Ig	B-DNA
variable	I-DNA
gene	I-DNA
families	I-DNA
,	O
are	O
required	O
for	O
normal	O
promoter	O
function	O
.	O

These	O
conserved	O
motifs	O
do	O
not	O
stimulate	O
transcription	O
in	O
the	O
absence	O
of	O
an	O
octamer	B-DNA
.	O

One	O
example	O
is	O
an	O
E-box	B-DNA
of	O
the	O
E47/E12	B-DNA
type	I-DNA
(	I-DNA
5'-CAGCTG-3	I-DNA
'	I-DNA
)	I-DNA
,	O
which	O
is	O
found	O
in	O
all	O
promoters	O
of	O
the	O
human	B-DNA
and	I-DNA
murine	I-DNA
Ig-kappa	I-DNA
gene	I-DNA
subgroups/families	I-DNA
,	O
with	O
the	O
exception	O
of	O
subgroups	O
II	O
and	O
VI	O
and	O
their	O
related	O
murine	B-DNA
families	I-DNA
.	O

In	O
the	O
present	O
study	O
we	O
show	O
that	O
the	O
ubiquitously	O
expressed	O
transcription	B-protein
factor	I-protein
AP-4	I-protein
,	O
and	O
not	O
E47	B-protein
,	O
interacts	O
specifically	O
with	O
the	O
kappa	B-DNA
promoter	I-DNA
E-boxes	I-DNA
when	O
tested	O
in	O
electrophoretic	O
mobility-shift	O
assays	O
using	O
nuclear	O
extracts	O
derived	O
from	O
human	B-cell_line
and	I-cell_line
murine	I-cell_line
B-cell	I-cell_line
lines	I-cell_line
.	O

Furthermore	O
,	O
AP-4	B-protein
,	O
unlike	O
E47	B-protein
,	O
did	O
not	O
act	O
as	O
a	O
transactivator	B-protein
,	O
which	O
is	O
in	O
agreement	O
with	O
previous	O
studies	O
on	O
intact	O
kappa	B-DNA
promoters	I-DNA
,	O
showing	O
that	O
transcription	O
is	O
absent	O
when	O
the	O
octamer	B-DNA
element	I-DNA
has	O
been	O
mutated	O
.	O

Based	O
on	O
these	O
data	O
,	O
and	O
the	O
conservation	O
of	O
the	O
5'-CAGCTG-3	B-DNA
'	I-DNA
motif	I-DNA
among	O
human	B-DNA
and	I-DNA
murine	I-DNA
kappa	I-DNA
promoters	I-DNA
,	O
we	O
propose	O
that	O
AP-4	B-protein
is	O
the	O
major	B-protein
ligand	I-protein
for	O
Ig-kappa	B-DNA
promoter	I-DNA
E-boxes	I-DNA
.	O

-DOCSTART-	O

A	O
16-mer	O
peptide	O
(	O
RQIKIWFQNRRMKWKK	O
)	O
from	O
antennapedia	O
preferentially	O
targets	O
the	O
Class	O
I	O
pathway	O
.	O

Translocation	O
of	O
antigenic	O
peptides	O
into	O
the	O
cytosol	O
of	O
antigen	B-cell_type
presenting	I-cell_type
cells	I-cell_type
facilitates	O
proteosomal	O
processing	O
and	O
loading	O
into	O
Class	B-protein
I	I-protein
molecules	I-protein
for	O
MHC	O
presentation	O
on	O
the	O
cell	O
surface	O
.	O

The	O
DNA	B-protein
binding	I-protein
domain	I-protein
of	O
the	O
Drosophila	B-protein
transcription	I-protein
factor	I-protein
(	O
Antennapedia	B-protein
)	O
,	O
a	O
60	B-protein
amino	I-protein
acid	I-protein
protein	I-protein
,	O
is	O
rapidly	O
taken	O
up	O
by	O
cells	O
and	O
has	O
been	O
fused	O
to	O
selected	O
antigens	B-protein
to	O
enhance	O
their	O
immunogenicity	O
.	O

We	O
now	O
demonstrate	O
that	O
a	O
16	O
amino	O
acid	O
peptide	O
from	O
antennapedia	O
can	O
facilitate	O
the	O
cytoplasmic	O
uptake	O
of	O
CTL	O
epitope	O
9-mer	O
peptides	O
.	O

Synthetic	O
peptides	O
were	O
made	O
containing	O
the	O
16-mer	O
antennapedia	O
peptide	O
linked	O
in	O
tandem	O
to	O
the	O
ovalbumin	O
SIINFEKL	O
CTL	O
peptide	O
.	O

The	O
peptide	O
complex	O
was	O
shown	O
to	O
rapidly	O
internalise	O
into	O
APCs	B-cell_type
by	O
confocal	O
microscopy	O
.	O

This	O
peptide	O
induced	O
CTL	B-cell_type
in	O
C57BL/6	O
mice	O
and	O
protected	O
them	O
against	O
growth	O
of	O
an	O
ovalbumin	B-cell_line
expressing	I-cell_line
tumour	I-cell_line
cell	I-cell_line
line	I-cell_line
(	O
E.G7-OVA	B-cell_line
)	O
.	O

The	O
ability	O
of	O
the	O
hybrid	O
peptide	O
to	O
be	O
processed	O
and	O
presented	O
by	O
APCs	B-cell_type
was	O
similar	O
,	O
whether	O
the	O
SIINFEKL	O
sequence	O
was	O
appended	O
at	O
the	O
C-terminus	B-protein
or	O
N-terminus	B-protein
of	O
the	O
Antennapedia	O
peptide	O
.	O

The	O
production	O
of	O
synthetic	O
peptides	O
containing	O
other	O
CTL	O
peptide	O
epitopes	O
may	O
be	O
useful	O
for	O
priming	O
CTLs	B-cell_type
in	O
vitro	O
and	O
in	O
vivo	O

-DOCSTART-	O

Bone	B-cell_type
marrow	I-cell_type
cells	I-cell_type
promote	O
TH2	O
polarization	O
and	O
inhibit	O
virus-specific	O
CTL	O
generation	O
.	O

This	O
laboratory	O
recently	O
reported	O
that	O
human	B-cell_type
bone	I-cell_type
marrow	I-cell_type
cells	I-cell_type
(	O
BMC	B-cell_type
)	O
inhibit	O
the	O
generation	O
of	O
virus-specific	B-cell_type
CTL	I-cell_type
in	O
culture	O
.	O

The	O
culture	O
supernatants	O
contained	O
increased	O
levels	O
of	O
prostaglandin	B-protein
E	I-protein
(	I-protein
2	I-protein
)	I-protein
(	I-protein
PGE	I-protein
(	I-protein
2	I-protein
)	I-protein
)	I-protein
(	O
shown	O
to	O
favor	O
TH2	O
cell	O
development	O
)	O
and	O
also	O
inhibited	O
EBV-CTL	O
effector	O
cell	O
development	O
.	O

In	O
this	O
study	O
,	O
we	O
obtained	O
PBL	B-cell_type
from	O
Epstein-Barr	O
virus	O
(	O
EBV	O
)	O
IgG	B-protein
antibody	I-protein
positive	O
kidney	O
transplant	O
recipients	O
(	O
R	O
)	O
and	O
their	O
living-related	O
donors	O
(	O
LRD	O
)	O
one	O
year	O
after	O
renal	O
transplantation	O
.	O

EBV-specific	B-cell_type
CTL	I-cell_type
were	O
then	O
generated	O
in	O
vitro	O
by	O
stimulating	O
PBL	B-cell_type
with	O
autologous	O
EBV-transformed	B-cell_type
B	I-cell_type
cells	I-cell_type
(	O
EBV-B	B-cell_type
)	O
in	O
the	O
presence	O
or	O
absence	O
of	O
autologous	B-cell_type
BMC	I-cell_type
.	O

The	O
addition	O
of	O
BMC	B-cell_type
to	O
the	O
EBV-CTL	B-cell_type
generation	I-cell_type
cultures	I-cell_type
increased	O
the	O
intracellular	O
expression	O
in	O
CD3+	B-cell_type
cells	I-cell_type
of	O
IL-4	B-protein
,	I-protein
-5	I-protein
,	I-protein
-6	I-protein
,	I-protein
-10	I-protein
,	I-protein
and	I-protein
-13	I-protein
.	O

These	O
CD3+	B-cell_type
cells	I-cell_type
also	O
expressed	O
increased	O
levels	O
of	O
the	O
TH2	B-protein
associated	I-protein
receptor	I-protein
CCR3	I-protein
.	O

Inhibition	O
was	O
even	O
observed	O
by	O
preparing	O
EBV-CTL	B-cell_type
generating	I-cell_type
cultures	I-cell_type
in	O
trans-wells	O
that	O
separated	O
the	O
autologous	B-cell_type
BMC	I-cell_type
from	O
the	O
PBL	B-cell_type
+	O
EBV-B	B-cell_type
.	O

It	O
was	O
then	O
observed	O
that	O
CD3+	B-cell_type
cells	I-cell_type
obtained	O
after	O
7	O
days	O
of	O
culture	O
in	O
the	O
presence	O
of	O
autologous	B-cell_type
BMC	I-cell_type
could	O
be	O
used	O
as	O
inhibitors	O
of	O
EBV-CTL	O
generation	O
.	O

Protein	B-protein
Kinase	I-protein
A	I-protein
(	O
PKA	B-protein
)	O
,	O
a	O
cAMP	B-protein
kinase	I-protein
that	O
is	O
involved	O
in	O
the	O
upregulation	O
of	O
TH2	B-protein
cytokine	I-protein
activity	O
,	O
was	O
increased	O
in	O
EBV-CTL	B-cell_type
cultures	I-cell_type
by	O
the	O
presence	O
of	O
BMC	B-cell_type
.	O

Additionally	O
,	O
IL-4-mediated	O
signal	O
transduction	O
and	O
activation	O
of	O
transcription	O
(	O
STAT-6	B-protein
)	O
phosphorylation	O
was	O
slightly	O
increased	O
.	O

These	O
results	O
show	O
that	O
the	O
BMC	B-cell_type
inhibition	O
is	O
mediated	O
by	O
soluble	B-protein
factors	I-protein
(	O
cytokines	B-protein
)	O
and	O
that	O
cell-cell	O
contact	O
in	O
this	O
autologous	O
system	O
is	O
not	O
required	O
,	O
so	O
that	O
BMC	B-cell_type
(	O
at	O
least	O
partially	O
,	O
via	O
cytokine	O
production	O
)	O
promote	O
TH2	O
polarization	O
in	O
culture	O
.	O

Moreover	O
,	O
TH2	B-cell_type
cells	I-cell_type
induced	O
by	O
culturing	O
with	O
autologous	B-cell_type
BMC	I-cell_type
directly	O
inhibit	O
EBV-CTL	O
generation	O
,	O
and	O
TH2	B-protein
associated	O
PKA	B-protein
,	O
CCR3	B-protein
,	O
and	O
STAT-6	B-protein
phosphorylation	O
are	O
enhanced	O
by	O
BMC	B-cell_type
.	O

-DOCSTART-	O

Reduction	O
in	O
DNA	O
binding	O
activity	O
of	O
the	O
transcription	B-protein
factor	I-protein
Pax-5a	B-protein
in	O
B	B-cell_type
lymphocytes	I-cell_type
of	O
aged	O
mice	O
.	O

Aging	O
has	O
been	O
associated	O
with	O
intrinsic	O
changes	O
of	O
the	O
humoral	O
immune	O
response	O
,	O
which	O
may	O
lead	O
to	O
an	O
increased	O
occurrence	O
of	O
autoimmune	O
disorders	O
and	O
pathogenic	O
susceptibility	O
.	O

The	O
transcription	B-protein
factor	I-protein
Pax-5	B-protein
is	O
a	O
key	O
regulator	O
of	O
B	O
cell	O
development	O
.	O

Pax-5a/B	B-protein
cell-specific	I-protein
activator	I-protein
protein	I-protein
and	O
an	O
alternatively	B-protein
spliced	I-protein
isoform	I-protein
,	I-protein
Pax-5d	I-protein
,	O
may	O
have	O
opposing	O
functions	O
in	O
transcriptional	O
regulation	O
due	O
to	O
the	O
lack	O
of	O
a	O
transactivation	B-protein
domain	I-protein
in	O
Pax-5d	B-protein
.	O

To	O
study	O
B	B-cell_type
cell	I-cell_type
-specific	O
changes	O
that	O
occur	O
during	O
the	O
aging	O
process	O
,	O
we	O
investigated	O
expression	O
patterns	O
of	O
Pax-5a	B-protein
and	I-protein
5d	I-protein
in	O
mature	B-cell_type
B	I-cell_type
cells	I-cell_type
of	O
young	O
and	O
aged	O
mice	O
.	O

RNase	O
protection	O
assays	O
showed	O
a	O
similar	O
transcriptional	O
pattern	O
for	O
both	O
age	O
groups	O
that	O
indicates	O
that	O
aging	O
has	O
no	O
affect	O
on	O
transcription	O
initiation	O
or	O
alternative	O
splicing	O
for	O
either	O
isoform	B-protein
.	O

In	O
contrast	O
,	O
a	O
significant	O
reduction	O
in	O
the	O
DNA	O
binding	O
activity	O
of	O
Pax-5a	B-protein
but	O
not	O
Pax-5d	B-protein
protein	O
was	O
observed	O
in	O
aged	B-cell_type
B	I-cell_type
cells	I-cell_type
in	O
vitro	O
,	O
while	O
Western	O
blot	O
analyses	O
showed	O
that	O
similar	O
levels	O
of	O
Pax-5a	B-protein
and	I-protein
5d	I-protein
proteins	O
were	O
present	O
in	O
both	O
age	O
groups	O
.	O

The	O
observed	O
decrease	O
in	O
Pax-5a	B-protein
binding	O
activity	O
correlated	O
with	O
changes	O
in	O
expression	O
of	O
two	O
Pax-5	B-DNA
target	I-DNA
genes	I-DNA
in	O
aged	B-cell_type
B	I-cell_type
cells	I-cell_type
.	O

Expression	O
of	O
the	O
Ig	B-protein
J	I-protein
chain	I-protein
and	O
the	O
secreted	O
form	O
of	O
Ig	B-protein
mu	I-protein
,	O
which	O
are	O
both	O
known	O
to	O
be	O
suppressed	O
by	O
Pax-5a	B-protein
in	O
mature	B-cell_type
B	I-cell_type
cells	I-cell_type
,	O
were	O
increased	O
in	O
B	B-cell_type
cells	I-cell_type
of	O
aged	O
mice	O
.	O

Together	O
,	O
our	O
studies	O
suggest	O
that	O
changes	O
associated	O
with	O
the	O
aging	O
phenotype	O
cause	O
posttranslational	O
modification	O
(	O
s	O
)	O
of	O
Pax-5a	B-protein
but	O
not	O
Pax-5d	B-protein
,	O
which	O
may	O
lead	O
to	O
an	O
abnormal	O
B	O
cell	O
phenotype	O
in	O
aged	O
mice	O
,	O
associated	O
with	O
elevated	O
levels	O
of	O
J	B-protein
chain	I-protein
,	O
and	O
secretion	O
of	O
IgM	B-protein

-DOCSTART-	O

NF-kappa	B-protein
B/Rel	I-protein
participation	O
in	O
the	O
lymphokine-dependent	O
proliferation	O
of	O
T	B-cell_type
lymphoid	I-cell_type
cells	I-cell_type
.	O

Proliferative	O
responses	O
of	O
lymphoid	B-cell_type
cells	I-cell_type
to	O
IL-2	B-protein
and	O
IL-4	B-protein
depend	O
on	O
activation	O
of	O
the	O
cells	O
,	O
but	O
the	O
mechanism	O
(	O
s	O
)	O
by	O
which	O
activation	O
enhances	O
cellular	O
competence	O
to	O
respond	O
to	O
cytokines	B-protein
is	O
not	O
fully	O
understood	O
.	O

The	O
NF-kappaB/Rel	B-protein
family	I-protein
represents	O
one	O
signal	O
transduction	O
pathway	O
induced	O
during	O
such	O
activation	O
.	O

We	O
show	O
in	O
this	O
study	O
that	O
inhibition	O
of	O
NF-kappaB	B-protein
through	O
the	O
expression	O
of	O
an	O
IkappaBalpha	B-protein
(	O
inhibitory	B-protein
protein	I-protein
that	O
dissociates	O
from	O
NF-kappaB	B-protein
)	O
mutant	O
refractory	O
to	O
signal-induced	O
degradation	O
(	O
IkappaBalpha	B-protein
(	I-protein
DeltaN	I-protein
)	I-protein
)	O
interfered	O
with	O
the	O
acquisition	O
of	O
competence	O
to	O
proliferate	O
in	O
response	O
to	O
IL-4	B-protein
as	O
well	O
as	O
IL-2	B-protein
.	O

Thymocytes	B-cell_type
and	O
T	B-cell_type
cells	I-cell_type
from	O
IkappaBalpha	B-protein
(	I-protein
DeltaN	I-protein
)	I-protein
transgenic	O
mice	O
expressed	O
normal	O
levels	O
of	O
IL-2R	B-protein
subunits	I-protein
.	O

However	O
,	O
transgenic	B-cell_type
cells	I-cell_type
exhibited	O
a	O
dramatic	O
defect	O
in	O
Stat5A	O
activation	O
treatment	O
with	O
IL-2	B-protein
,	O
and	O
a	O
similar	O
defect	O
was	O
observed	O
for	O
IL-4	B-protein
-induced	O
Stat5	B-protein
.	O

In	O
contrast	O
,	O
T	B-cell_type
lymphoid	I-cell_type
cells	I-cell_type
with	O
inhibition	O
of	O
NF-kappaB	B-protein
showed	O
normal	O
insulin	O
receptor	O
substrate-2	O
phosphorylation	O
and	O
only	O
a	O
modest	O
decrease	O
in	O
Stat6	O
activation	O
and	O
insulin	O
receptor	O
substrate-1	O
phosphorylation	O
after	O
IL-4	O
stimulation	O
.	O

These	O
results	O
indicate	O
that	O
the	O
NF-kappaB/Rel/IkappaBalpha	O
system	O
can	O
regulate	O
cytokine	B-protein
receptor	I-protein
capacitation	O
through	O
effects	O
on	O
the	O
induction	O
of	O
downstream	O
signaling	O
by	O
the	O
Stat	B-protein
transcription	I-protein
factor	I-protein
family	I-protein
.	O

-DOCSTART-	O

BMS-189453	O
,	O
a	O
novel	O
retinoid	O
receptor	O
antagonist	O
,	O
is	O
a	O
potent	O
testicular	O
toxin	O
.	O

BMS-189453	O
is	O
a	O
synthetic	O
retinoid	O
that	O
acts	O
as	O
an	O
antagonist	O
at	O
retinoic	B-protein
acid	I-protein
receptors	I-protein
alpha	I-protein
,	I-protein
beta	I-protein
,	I-protein
and	I-protein
gamma	I-protein
.	O

In	O
Sprague	O
Dawley	O
rats	O
at	O
daily	O
oral	O
doses	O
of	O
15	O
,	O
60	O
,	O
or	O
240	O
mg/kg	O
for	O
1	O
month	O
,	O
BMS-189453	O
produced	O
increases	O
in	O
leukocyte	B-cell_type
counts	O
,	O
alkaline	B-protein
phosphatase	I-protein
and	O
alanine	B-protein
aminotransferase	I-protein
levels	O
,	O
and	O
marked	O
testicular	O
degeneration	O
and	O
atrophy	O
at	O
all	O
doses	O
.	O

Significant	O
overt	O
signs	O
of	O
toxicity	O
and	O
deaths	O
occurred	O
at	O
240	O
mg/kg	O
,	O
whereas	O
body-weight	O
and	O
food-consumption	O
decreases	O
occurred	O
at	O
60	O
and	O
240	O
mg/kg	O
.	O

When	O
BMS-189453	O
was	O
administered	O
to	O
male	O
rats	O
at	O
daily	O
doses	O
ranging	O
from	O
12.5	O
to	O
100	O
mg/kg	O
for	O
1	O
week	O
,	O
only	O
minimal	O
testicular	O
changes	O
occurred	O
at	O
all	O
doses	O
,	O
shortly	O
after	O
the	O
dosing	O
period	O
.	O

However	O
,	O
after	O
a	O
1-month	O
drug-free	O
observation	O
period	O
,	O
marked	O
testicular	O
atrophy	O
was	O
evident	O
at	O
all	O
doses	O
.	O

BMS-189453	O
was	O
then	O
administered	O
at	O
doses	O
of	O
2	O
,	O
10	O
,	O
or	O
50	O
mg/kg	O
to	O
male	O
rats	O
for	O
1	O
,	O
3	O
,	O
or	O
7	O
consecutive	O
days	O
.	O

Dose-	O
and	O
duration-dependent	O
testicular	O
toxicity	O
that	O
occurred	O
after	O
a	O
1-month	O
observation	O
period	O
did	O
not	O
recover	O
,	O
and	O
,	O
in	O
some	O
cases	O
,	O
was	O
more	O
severe	O
4	O
months	O
after	O
the	O
last	O
dose	O
.	O

In	O
rabbits	O
administered	O
BMS-189453	O
at	O
oral	O
doses	O
of	O
2	O
,	O
10	O
,	O
or	O
50	O
mg/kg	O
for	O
1	O
week	O
,	O
testicular	O
degeneration	O
and	O
atrophy	O
were	O
evident	O
in	O
the	O
high-dose	O
group	O
at	O
1	O
month	O
following	O
treatment	O
.	O

These	O
studies	O
indicate	O
that	O
retinoid	O
antagonists	O
can	O
selectively	O
produce	O
progressive	O
and	O
prolonged	O
testicular	O
toxicity	O
after	O
single	O
or	O
repeated	O
oral	O
doses	O
that	O
are	O
otherwise	O
well	O
tolerated	O
.	O

-DOCSTART-	O

Sequential	O
involvement	O
of	O
NFAT	B-protein
and	O
Egr	B-protein
transcription	B-protein
factors	I-protein
in	O
FasL	O
regulation	O
.	O

The	O
critical	O
function	O
of	O
NFAT	B-protein
proteins	I-protein
in	O
maintaining	O
lymphoid	O
homeostasis	O
was	O
revealed	O
in	O
mice	O
lacking	O
both	O
NFATp	B-protein
and	O
NFAT4	B-protein
(	O
DKO	O
)	O
.	O

DKO	O
mice	O
exhibit	O
increased	O
lymphoproliferation	O
,	O
decreased	O
activation-induced	O
cell	O
death	O
,	O
and	O
impaired	O
induction	O
of	O
FasL	B-protein
.	O

The	O
transcription	B-protein
factors	I-protein
Egr2	B-protein
and	O
Egr3	B-protein
are	O
potent	O
activators	O
of	O
FasL	O
expression	O
.	O

Here	O
we	O
find	O
that	O
Egr2	B-DNA
and	O
Egr3	B-DNA
are	O
NFAT	B-DNA
target	I-DNA
genes	I-DNA
.	O

Activation	O
of	O
FasL	B-protein
occurs	O
via	O
the	O
NFAT	B-protein
-dependent	O
induction	O
of	O
Egr3	B-DNA
,	O
as	O
demonstrated	O
by	O
the	O
ability	O
of	O
exogenously	O
provided	O
NFATp	B-protein
to	O
restore	O
Egr	B-protein
-dependent	O
FasL	O
promoter	O
activity	O
in	O
DKO	B-cell_type
lymph	I-cell_type
node	I-cell_type
cells	I-cell_type
.	O

Further	O
,	O
Egr3	B-DNA
expression	O
is	O
enriched	O
in	O
Th1	B-cell_type
cells	I-cell_type
,	O
suggesting	O
a	O
molecular	O
basis	O
for	O
the	O
known	O
preferential	O
expression	O
of	O
FasL	B-protein
in	O
the	O
Th1	O
versus	O
Th2	O
subset	O
.	O

-DOCSTART-	O

Mechanisms	O
and	O
clinical	O
relevance	O
of	O
nongenomic	O
glucocorticoid	O
actions	O
.	O

Glucocorticoids	O
have	O
profound	O
anti-inflammatory	O
and	O
immunosuppressive	O
actions	O
when	O
used	O
therapeutically	O
.	O

The	O
therapeutic	O
dose	O
is	O
quite	O
variable	O
and	O
depends	O
on	O
the	O
disease	O
,	O
but	O
ranges	O
from	O
very	O
low	O
to	O
extremely	O
high	O
.	O

The	O
rationale	O
for	O
the	O
use	O
of	O
various	O
dosage	O
regimens	O
for	O
specific	O
clinical	O
indications	O
is	O
the	O
existence	O
of	O
three	O
distinct	O
,	O
therapeutically	O
relevant	O
effects	O
:	O
genomic	O
,	O
specific	O
nongenomic	O
and	O
unspecific	O
nongenomic	O
.	O

Genomic	O
effects	O
are	O
mediated	O
by	O
cytosolic	B-protein
receptors	I-protein
that	O
alter	O
expression	O
of	O
specific	O
genes	O
.	O

Specific	O
nongenomic	O
effects	O
occur	O
within	O
a	O
few	O
minutes	O
and	O
are	O
mediated	O
by	O
steroid-selective	B-protein
membrane	I-protein
receptors	I-protein
.	O

Unspecific	O
nongenomic	O
effects	O
occur	O
within	O
seconds	O
,	O
but	O
only	O
at	O
high	O
glucocorticoid	O
dosages	O
,	O
and	O
seem	O
to	O
result	O
from	O
direct	O
interactions	O
with	O
biological	O
membranes	O
.	O

For	O
unspecific	O
nongenomic	O
effects	O
,	O
methylprednisolone	O
and	O
other	O
glucocorticoids	O
have	O
been	O
shown	O
to	O
inhibit	O
cation	O
cycling	O
across	O
the	O
plasma	O
membrane	O
,	O
but	O
to	O
have	O
little	O
effect	O
on	O
protein	O
synthesis	O
.	O

Thus	O
,	O
glucocorticoids	O
could	O
diminish	O
or	O
prevent	O
the	O
acute	O
immune	O
response	O
by	O
interfering	O
with	O
processes	O
such	O
as	O
the	O
rise	O
in	O
intracellular	O
Ca2+	O
concentration	O
.	O

It	O
is	O
proposed	O
that	O
the	O
additional	O
therapeutic	O
benefit	O
of	O
higher	O
doses	O
is	O
obtained	O
via	O
these	O
nongenomic	O
effects	O
.	O

-DOCSTART-	O

High	O
glucose-induced	O
intercellular	B-protein
adhesion	I-protein
molecule-1	I-protein
(	O
ICAM-1	B-protein
)	O
expression	O
through	O
an	O
osmotic	O
effect	O
in	O
rat	B-cell_type
mesangial	I-cell_type
cells	I-cell_type
is	O
PKC-NF-kappa	O
B-dependent	O
.	O

AIMS/HYPOTHESIS	O
:	O
Infiltration	O
of	O
mononuclear	B-cell_type
cells	I-cell_type
and	O
glomerular	O
enlargement	O
accompanied	O
by	O
glomerular	B-cell_type
cell	I-cell_type
proliferation	O
are	O
very	O
early	O
characteristics	O
of	O
the	O
pathophysiology	O
of	O
diabetes	O
.	O

To	O
clarify	O
the	O
mechanism	O
of	O
early	O
diabetic	O
nephropathy	O
,	O
we	O
measured	O
[	O
3H	O
]	O
-thymidine	O
incorporation	O
and	O
cell	O
numbers	O
to	O
show	O
the	O
influence	O
of	O
a	O
high	O
ambient	O
glucose	O
concentration	O
and	O
the	O
osmotic	O
effect	O
on	O
rat	B-cell_type
mesangial	I-cell_type
cell	I-cell_type
proliferation	O
.	O

We	O
also	O
measured	O
the	O
effect	O
of	O
high	O
glucose	O
on	O
the	O
expression	O
of	O
intercellular	B-protein
adhesion	I-protein
molecule-1	I-protein
and	O
vascular	B-protein
adhesion	I-protein
molecule-1	I-protein
by	O
flow	O
cytometry	O
and	O
semiquantitative	O
RT-PCR	O
in	O
mesangial	B-cell_type
cells	I-cell_type
and	O
the	O
adhesion	O
of	O
leukocytes	B-cell_type
to	O
mesangial	B-cell_type
cells	I-cell_type
.	O

METHODS/RESULTS	O
:	O
Cells	O
exposed	O
to	O
high	O
D-glucose	O
(	O
30	O
mmol/l	O
)	O
caused	O
an	O
increase	O
in	O
[	O
3H	O
]	O
-thymidine	O
incorporation	O
and	O
cell	O
numbers	O
at	O
24	O
and	O
48	O
h	O
and	O
normalized	O
at	O
72	O
h	O
(	O
p	O
<	O
0.05	O
)	O
,	O
whereas	O
these	O
changes	O
were	O
not	O
found	O
in	O
high	O
mannitol	O
(	O
30	O
mmol/l	O
)	O
,	O
IL-1	B-protein
beta	I-protein
,	O
or	O
TNF	B-protein
alpha	I-protein
-stimulated	O
mesangial	B-cell_type
cells	I-cell_type
.	O

Cells	O
exposed	O
to	O
high-glucose	O
(	O
15	O
,	O
30	O
,	O
or	O
60	O
mmol/l	O
)	O
or	O
osmotic	O
agents	O
(	O
L-glucose	O
,	O
raffinose	O
and	O
mannitol	O
)	O
showed	O
that	O
intercellular	B-protein
adhesion	I-protein
molecule-1	I-protein
expression	O
began	O
to	O
increase	O
after	O
24	O
h	O
,	O
reached	O
its	O
maximum	O
at	O
24	O
and	O
48	O
h	O
and	O
gradually	O
decreased	O
afterwards	O
.	O

The	O
stimulatory	O
effects	O
of	O
high	O
glucose	O
and	O
high	O
mannitol	O
on	O
mRNA	O
expression	O
were	O
observed	O
as	O
early	O
as	O
6	O
h	O
and	O
reached	O
its	O
maximum	O
at	O
12	O
h	O
.	O

Up-regulation	O
of	O
ICAM-1	B-protein
protein	O
and	O
mRNA	O
was	O
also	O
found	O
in	O
IL-1-beta	B-protein
and	O
TNF-alpha	B-protein
-stimulated	O
mesangial	B-cell_type
cells	I-cell_type
.	O

Neither	O
vascular	B-protein
adhesion	I-protein
molecule-1	I-protein
protein	O
nor	O
mRNA	O
expression	O
was	O
,	O
however	O
,	O
affected	O
by	O
high	O
glucose	O
and	O
high	O
mannitol	O
.	O

Notably	O
,	O
the	O
protein	O
kinase	O
C	O
inhibitors	O
calphostin	O
C	O
and	O
staurosporine	O
reduced	O
high	O
glucose-	O
or	O
high	O
mannitol-induced	O
intercellular	B-RNA
adhesion	I-RNA
molecule-1	I-RNA
mRNA	O
expression	O
and	O
high	O
glucose-induced	O
proliferation	O
.	O

Furthermore	O
,	O
the	O
NF-kappa	O
B	O
inhibitor	O
N-tosyl-L-phenylalanine	O
chloromethyl	O
ketone	O
reduced	O
high	O
glucose-	O
or	O
high	O
mannitol-induced	O
intercellular	B-RNA
adhesion	I-RNA
molecule-1	I-RNA
mRNA	O
expression	O
and	O
high	O
glucose-induced	O
proliferation	O
.	O

Results	O
showed	O
that	O
high	O
glucose	O
(	O
15	O
,	O
30	O
mmol/l	O
)	O
or	O
high	O
concentrations	O
of	O
osmotic	O
agents	O
remarkably	O
increased	O
the	O
number	O
of	O
adherent	O
leukocytes	B-cell_type
to	O
mesangial	B-cell_type
cells	I-cell_type
(	O
p	O
<	O
0.01	O
)	O
compared	O
with	O
control	B-cell_type
cells	I-cell_type
(	O
5	O
mmol/l	O
D-glucose	O
)	O
.	O

Functional	O
blocking	O
of	O
intercellular	B-protein
adhesion	I-protein
molecule-1	I-protein
on	O
mesangial	B-cell_type
cells	I-cell_type
with	O
rat	B-protein
intercellular	I-protein
adhesion	I-protein
molecule-1	I-protein
monoclonal	I-protein
antibody	I-protein
,	O
calphostin	O
C	O
,	O
staurosporine	O
,	O
or	O
N-tosyl-L-phenylalanine	O
chloromethyl	O
ketone	O
significantly	O
inhibited	O
high	O
glucose-	O
or	O
high	O
mannitol-induced	O
increase	O
in	O
leukocyte	O
adhesion	O
(	O
p	O
<	O
<	O
0.05	O
)	O
.	O

CONCLUSION/INTERPRETATION	O
:	O
These	O
results	O
suggest	O
that	O
high	O
glucose	O
can	O
upregulate	O
intercellular	B-protein
adhesion	I-protein
molecule-1	I-protein
protein	O
and	O
mRNA	O
expression	O
but	O
not	O
vascular	O
adhesion	O
molecule-1	O
expression	O
in	O
mesangial	B-cell_type
cells	I-cell_type
and	O
promote	O
leukocyte	O
adhesion	O
through	O
up-regulation	O
of	O
intercellular	B-protein
adhesion	I-protein
molecule-1	I-protein
through	O
osmotic	O
effect	O
,	O
possibly	O
depending	O
on	O
the	O
protein	B-protein
kinase	I-protein
C	I-protein
nuclear	B-protein
factor-kappa	I-protein
B	I-protein
(	O
PKC	B-protein
-	O
NF-kappa	B-protein
B	I-protein
)	O
pathway	O
.	O

High	O
glucose	O
itself	O
can	O
also	O
promote	O
mesangial	O
cell	O
proliferation	O
through	O
the	O
PKC-NF-kappa	O
B	O
pathways	O
.	O

We	O
conclude	O
that	O
hyperglycaemia	O
in	O
itself	O
seems	O
to	O
be	O
an	O
important	O
factor	O
in	O
the	O
development	O
of	O
early	O
diabetic	O
nephropathy	O
.	O

-DOCSTART-	O

Combined	O
corticosteroid/	O
granulocyte	B-protein
colony-stimulating	I-protein
factor	I-protein
(	O
G-CSF	B-protein
)	O
therapy	O
in	O
the	O
treatment	O
of	O
severe	O
congenital	O
neutropenia	O
unresponsive	O
to	O
G-CSF	B-protein
:	O
Activated	O
glucocorticoid	B-protein
receptors	I-protein
synergize	O
with	O
G-CSF	B-protein
signals	O
.	O

More	O
than	O
90	O
%	O
of	O
patients	O
with	O
severe	O
congenital	O
neutropenia	O
(	O
SCN	O
)	O
respond	O
to	O
granulocyte	B-protein
colony-stimulating	I-protein
factor	I-protein
(	O
G-CSF	B-protein
)	O
therapy	O
.	O

The	O
basis	O
for	O
the	O
refractory	O
state	O
in	O
the	O
remaining	O
patients	O
is	O
unknown	O
.	O

To	O
address	O
this	O
issue	O
,	O
we	O
studied	O
a	O
child	O
with	O
SCN	O
who	O
was	O
totally	O
unresponsive	O
to	O
G-CSF	B-protein
and	O
had	O
a	O
novel	O
point	O
mutation	O
in	O
the	O
extracellular	B-protein
domain	I-protein
of	O
the	O
G-CSF	B-protein
receptor	I-protein
(	O
GCSF-R	B-protein
)	O
.	O

Marrow	O
stromal	O
support	O
of	O
granulopoiesis	O
was	O
evaluated	O
by	O
plating	O
CD34	B-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
cells	I-cell_type
on	O
preformed	O
stromal	O
layers	O
.	O

Nonadherent	B-cell_type
cells	I-cell_type
were	O
harvested	O
and	O
assayed	O
in	O
clonogenic	O
assays	O
for	O
granulocytic	O
colony	O
production	O
.	O

The	O
in	O
vitro	O
effect	O
of	O
G-CSF	B-protein
and	O
corticosteroids	O
on	O
granulopoiesis	O
was	O
evaluated	O
in	O
clonogenic	O
assays	O
of	O
marrow	B-cell_type
mononuclear	I-cell_type
cells	I-cell_type
,	O
by	O
proliferation	O
studies	O
of	O
the	O
murine	B-cell_line
myeloid	I-cell_line
cell	I-cell_line
line	I-cell_line
32D	I-cell_line
expressing	O
the	O
patient	O
's	O
mutated	B-protein
G-CSFR	I-protein
,	O
and	O
by	O
measuring	O
STAT5	O
activation	O
in	O
nuclear	O
extracts	O
from	O
stimulated	B-cell_type
cells	I-cell_type
.	O

Patient	O
's	O
stroma	O
supported	O
granulopoiesis	O
derived	O
from	O
control	O
marrow	B-cell_type
CD34	I-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
cells	I-cell_type
in	O
a	O
normal	O
manner	O
.	O

Normal	O
stroma	O
,	O
however	O
,	O
failed	O
to	O
induce	O
granulopoiesis	O
from	O
patient	O
's	O
CD34	B-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
cells	I-cell_type
.	O

Clonogenic	O
assays	O
of	O
the	O
patient	O
's	O
marrow	B-cell_type
mononuclear	I-cell_type
cells	I-cell_type
incorporating	O
either	O
G-CSF	B-protein
or	O
hydrocortisone	O
produced	O
little	O
neutrophil	O
growth	O
.	O

In	O
contrast	O
,	O
inclusion	O
of	O
both	O
G-CSF	B-protein
and	O
hydrocortisone	O
in	O
the	O
cytokine	B-protein
``	I-protein
cocktail	I-protein
''	I-protein
markedly	O
increased	O
the	O
neutrophil	O
numbers	O
.	O

Proliferation	O
of	O
32D	B-cell_line
cells	I-cell_line
expressing	O
the	O
mutated	B-protein
receptor	I-protein
and	O
STAT5	O
activation	O
were	O
improved	O
by	O
a	O
combination	O
of	O
G-CSF	B-protein
and	O
dexamethasone	O
.	O

When	O
small	O
daily	O
doses	O
of	O
oral	O
prednisone	O
were	O
then	O
administered	O
to	O
the	O
patient	O
with	O
conventional	O
doses	O
of	O
subcutaneous	B-protein
G-CSF	I-protein
,	O
the	O
patient	O
responded	O
with	O
increased	O
neutrophil	O
numbers	O
and	O
with	O
a	O
complete	O
reversal	O
of	O
the	O
infectious	O
problems	O
.	O

These	O
data	O
provide	O
insight	O
into	O
SCN	O
unresponsive	O
to	O
standard	O
G-CSF	B-protein
treatment	O
and	O
to	O
the	O
potential	O
corrective	O
action	O
of	O
combined	O
treatment	O
with	O
G-CSF	B-protein
and	O
corticosteroids	O
through	O
synergistic	O
activation	O
of	O
STAT5	B-protein
.	O

-DOCSTART-	O

Human	B-cell_type
renal	I-cell_type
mesangial	I-cell_type
cells	I-cell_type
are	O
a	O
target	O
for	O
the	O
anti-inflammatory	O
action	O
of	O
9-cis	O
retinoic	O
acid	O
.	O

Mesangial	B-cell_type
cells	I-cell_type
play	O
an	O
active	O
role	O
in	O
the	O
inflammatory	O
response	O
to	O
glomerular	O
injury	O
.	O

We	O
have	O
studied	O
in	O
cultured	B-cell_line
human	I-cell_line
mesangial	I-cell_line
cells	I-cell_line
(	O
CHMC	B-cell_line
)	O
several	O
effects	O
of	O
9-cis	O
retinoic	O
acid	O
(	O
9-cRA	O
)	O
,	O
an	O
activator	O
of	O
both	O
retinoic	B-protein
acid	I-protein
receptors	I-protein
(	O
RARs	B-protein
)	O
and	O
retinoid	B-protein
X	I-protein
receptors	I-protein
(	O
RXRs	B-protein
)	O
.	O

9-cRA	O
inhibited	O
foetal	O
calf	O
serum-induced	O
CHMC	B-cell_line
proliferation	O
.	O

It	O
also	O
prevented	O
CHMC	B-cell_line
death	O
induced	O
by	O
the	O
inflammatory	O
mediator	O
H	O
(	O
2	O
)	O
O	O
(	O
2	O
)	O
.	O

This	O
preventive	O
effect	O
was	O
not	O
due	O
to	O
any	O
increase	O
in	O
H	O
(	O
2	O
)	O
O	O
(	O
2	O
)	O
catabolism	O
and	O
it	O
persisted	O
even	O
when	O
both	O
catalase	O
and	O
glutathione	O
synthesis	O
were	O
inhibited	O
.	O

Finally	O
,	O
9-cRA	O
diminished	O
monocyte	O
adhesion	O
to	O
FCS-stimulated	B-cell_line
CHMC	I-cell_line
.	O

Interestingly	O
,	O
the	O
retinoid	O
also	O
inhibited	O
in	O
FCS-stimulated	B-cell_type
cells	I-cell_type
the	O
protein	O
expression	O
of	O
two	O
mesangial	B-protein
adhesion	I-protein
molecules	I-protein
,	O
fibronectin	B-protein
and	O
osteopontin	B-protein
,	O
but	O
it	O
did	O
not	O
modify	O
the	O
protein	O
expression	O
of	O
intercellular	B-protein
adhesion	I-protein
molecule-1	I-protein
and	O
vascular	B-protein
adhesion	I-protein
molecule-1	I-protein
.	O

All	O
major	O
RARs	B-protein
and	I-protein
RXRs	I-protein
isotypes	I-protein
were	O
expressed	O
in	O
CHMC	B-cell_line
regardless	O
of	O
the	O
presence	O
or	O
absence	O
of	O
9-cRA	O
.	O

Transcripts	O
to	O
RAR-alpha	B-RNA
,	O
RAR-beta	B-RNA
and	O
RXR-alpha	B-RNA
increased	O
after	O
incubation	O
with	O
9-cRA	O
whereas	O
RXR-gamma	B-RNA
was	O
inhibited	O
,	O
suggesting	O
a	O
major	O
role	O
for	O
RARs	B-protein
and	O
RXRs	B-protein
in	O
9-cRA-anti-inflammatory	O
effects	O
.	O

9-cRA	O
was	O
toxic	O
only	O
at	O
50	O
microM	O
(	O
a	O
concentration	O
50	O
-	O
5000	O
times	O
higher	O
than	O
required	O
for	O
the	O
effects	O
above	O
)	O
.	O

Cell	O
death	O
occurred	O
by	O
apoptosis	O
,	O
whose	O
onset	O
was	O
associated	O
with	O
a	O
pronounced	O
increase	O
in	O
catalase	O
activity	O
and	O
reduced	O
glutathione	O
content	O
,	O
being	O
more	O
effectively	O
induced	O
by	O
all-trans	O
retinoic	O
acid	O
.	O

Modulation	O
of	O
the	O
oxidant/antioxidant	O
balance	O
failed	O
to	O
inhibit	O
apoptosis	O
.	O

We	O
conclude	O
that	O
mesangial	B-cell_type
cells	I-cell_type
might	O
be	O
a	O
target	O
for	O
the	O
treatment	O
of	O
inflammatory	O
glomerulopathies	O
with	O
9-cRA	O
.	O

-DOCSTART-	O

Expression	O
of	O
mammalian	B-DNA
defensin	I-DNA
genes	I-DNA
.	O

Antimicrobial	O
peptides	O
are	O
a	O
prevalent	O
mechanism	O
of	O
host	O
defense	O
found	O
throughout	O
nature	O
.	O

In	O
mammals	O
,	O
defensins	B-protein
are	O
among	O
the	O
most	O
abundant	O
of	O
these	O
broad-spectrum	O
antibiotics	O
,	O
and	O
are	O
expressed	O
in	O
epithelial	B-cell_type
and	I-cell_type
hematopoietic	I-cell_type
cells	I-cell_type
.	O

The	O
defensin	O
peptides	O
are	O
especially	O
abundant	O
in	O
neutrophils	B-cell_type
;	O
however	O
,	O
gene	O
expression	O
is	O
limited	O
to	O
the	O
promyelocyte	O
stage	O
.	O

In	O
epithelial	B-cell_type
cells	I-cell_type
,	O
defensin	B-DNA
genes	I-DNA
are	O
found	O
as	O
both	O
constitutively	O
expressed	O
and	O
inducible	O
.	O

Induction	O
has	O
been	O
observed	O
in	O
vitro	O
by	O
stimulation	O
with	O
bacterial	O
lipopolysaccharide	O
as	O
well	O
as	O
inflammatory	O
mediators	O
.	O

In	O
vivo	O
,	O
up-regulation	O
of	O
several	O
defensin	B-DNA
genes	I-DNA
occurs	O
in	O
both	O
infectious	O
and	O
inflammatory	O
states	O
.	O

Gene	O
regulation	O
occurs	O
via	O
signal	O
transduction	O
pathways	O
common	O
to	O
other	O
innate	O
immune	O
responses	O
,	O
utilizing	O
transcription	B-protein
factors	I-protein
such	O
as	O
nuclear	B-protein
factor	I-protein
(	I-protein
NF	I-protein
)	I-protein
-kappaB	I-protein
and	O
NF	B-protein
interleukin-6	I-protein
.	O

Together	O
,	O
the	O
data	O
suggest	O
a	O
broad-based	O
innate	O
host	O
defense	O
whereby	O
potent	O
antimicrobial	O
peptides	O
are	O
present	O
to	O
prevent	O
initial	O
colonization	O
by	O
pathogenic	O
microorganisms	O
.	O

In	O
addition	O
,	O
the	O
recognition	O
of	O
bacteria	O
coupled	O
with	O
a	O
nascent	O
inflammatory	O
response	O
can	O
bolster	O
this	O
defense	O
by	O
a	O
coordinated	O
up-regulation	O
of	O
the	O
peptides	O
.	O

-DOCSTART-	O

Multiple	O
signals	O
required	O
for	O
cyclic	B-protein
AMP-responsive	I-protein
element	I-protein
binding	I-protein
protein	I-protein
(	O
CREB	B-protein
)	O
binding	O
protein	O
interaction	O
induced	O
by	O
CD3/CD28	O
costimulation	O
.	O

The	O
optimal	O
activation	O
of	O
cAMP-responsive	B-protein
element	I-protein
binding	I-protein
protein	I-protein
(	O
CREB	B-protein
)	O
,	O
similar	O
to	O
the	O
full	O
activation	O
of	O
T	B-cell_type
lymphocytes	I-cell_type
,	O
requires	O
the	O
stimulation	O
of	O
both	O
CD3	B-protein
and	O
CD28	B-protein
.	O

Using	O
a	O
reporter	O
system	O
to	O
detect	O
interaction	O
of	O
CREB	B-protein
and	O
CREB-binding	B-protein
protein	I-protein
(	O
CBP	B-protein
)	O
,	O
in	O
this	O
study	O
we	O
found	O
that	O
CREB	B-protein
binds	O
to	O
CBP	B-protein
only	O
by	O
engagement	O
of	O
both	O
CD3	B-protein
and	O
CD28	B-protein
.	O

CD3/CD28	B-protein
-promoted	O
CREB	B-protein
-	O
CBP	B-protein
interaction	O
was	O
dependent	O
on	O
p38	B-protein
mitogen-activated	I-protein
protein	I-protein
kinase	I-protein
(	O
MAPK	B-protein
)	O
and	O
calcium/calmodulin-dependent	B-protein
protein	I-protein
kinase	I-protein
(	I-protein
CaMK	I-protein
)	I-protein
IV	I-protein
in	O
addition	O
to	O
the	O
previously	O
identified	O
extracellular	O
signal-regulated	O
kinase	O
pathway	O
.	O

Extracellular	B-protein
signal-regulated	I-protein
kinase	I-protein
,	I-protein
CaMKIV	I-protein
,	O
and	O
p38	B-protein
MAPK	I-protein
were	O
also	O
the	O
kinases	O
involved	O
in	O
CREB	B-protein
Ser	O
(	O
133	O
)	O
phosphorylation	O
induced	O
by	O
CD3/CD28	B-protein
.	O

A	O
reconstitution	O
experiment	O
illustrated	O
that	O
optimum	O
CREB	B-protein
-	O
CBP	B-protein
interaction	O
and	O
CREB	B-protein
trans-activation	O
were	O
attained	O
when	O
these	O
three	O
kinase	O
pathways	O
were	O
simultaneously	O
activated	O
in	O
T	B-cell_type
cells	I-cell_type
.	O

Our	O
results	O
demonstrate	O
that	O
coordinated	O
activation	O
of	O
different	O
kinases	B-protein
leads	O
to	O
full	O
activation	O
of	O
CREB	B-protein
.	O

Notably	O
,	O
CD28	O
ligation	O
activated	O
p38	B-protein
MAPK	I-protein
and	O
CaMKIV	B-protein
,	O
the	O
kinases	B-protein
stimulated	O
by	O
CD3	O
engagement	O
,	O
suggesting	O
that	O
CD28	B-protein
acts	O
by	O
increasing	O
the	O
activation	O
extent	O
of	O
p38	B-protein
MAPK	I-protein
and	O
CaMKIV	B-protein
.	O

These	O
results	O
support	O
the	O
model	O
of	O
a	O
minimum	O
activation	O
threshold	O
for	O
CREB	B-protein
-	O
CBP	B-protein
interaction	O
that	O
can	O
be	O
reached	O
only	O
when	O
both	O
CD3	B-protein
and	O
CD28	B-protein
are	O
stimulated	O
.	O

-DOCSTART-	O

The	O
murine	B-DNA
IL-2	I-DNA
promoter	I-DNA
contains	O
distal	B-DNA
regulatory	I-DNA
elements	I-DNA
responsive	O
to	O
the	O
Ah	B-protein
receptor	I-protein
,	O
a	O
member	O
of	O
the	O
evolutionarily	O
conserved	O
bHLH-PAS	B-protein
transcription	I-protein
factor	I-protein
family	I-protein
.	O

Signaling	O
through	O
the	O
TCR	B-protein
and	O
costimulatory	O
signals	O
primarily	O
control	O
transcription	O
of	O
the	O
IL-2	B-DNA
gene	I-DNA
in	O
naive	B-cell_type
T	I-cell_type
cells	I-cell_type
.	O

The	O
minimal	B-DNA
promoter	I-DNA
necessary	O
for	O
this	O
expression	O
lies	O
proximal	O
,	O
between	O
-300	O
and	O
the	O
transcription	B-DNA
start	I-DNA
site	I-DNA
.	O

We	O
had	O
previously	O
shown	O
that	O
activation	O
of	O
the	O
arylhydrocarbon	B-protein
receptor	I-protein
(	O
AHR	B-protein
)	O
,	O
a	O
member	O
of	O
the	O
bHLH-PAS	B-protein
family	I-protein
of	I-protein
transcription	I-protein
factors	I-protein
,	O
leads	O
to	O
increased	O
mRNA	O
expression	O
of	O
IL-2	B-RNA
in	O
murine	B-cell_type
fetal	I-cell_type
thymocytes	I-cell_type
.	O

The	O
AHR	B-protein
is	O
abundant	O
in	O
the	O
thymus	O
and	O
may	O
play	O
a	O
role	O
for	O
the	O
development	O
of	O
the	O
immune	O
system	O
.	O

Moreover	O
,	O
its	O
overactivation	O
by	O
chemicals	O
such	O
as	O
dioxins	O
leads	O
to	O
immunosuppression	O
and	O
thymic	O
involution	O
.	O

Binding	O
motifs	O
for	O
the	O
liganded	B-protein
AHR	I-protein
can	O
be	O
identified	O
in	O
the	O
distal	B-DNA
region	I-DNA
-1300	I-DNA
to	I-DNA
-800	I-DNA
of	O
the	O
mouse	B-DNA
IL-2	I-DNA
promoter	I-DNA
.	O

We	O
show	O
here	O
that	O
these	O
DNA	B-DNA
motifs	I-DNA
,	O
the	O
so-called	O
dioxin	B-DNA
response	I-DNA
elements	I-DNA
,	O
after	O
binding	O
to	O
the	O
liganded	B-protein
AHR	I-protein
are	O
sufficient	O
to	O
transactivate	O
luciferase	O
expression	O
in	O
a	O
reporter	O
gene	O
system	O
.	O

The	O
IL-2	B-DNA
gene	I-DNA
can	O
be	O
induced	O
by	O
the	O
AHR	B-protein
also	O
in	O
thymocytes	B-cell_type
in	O
vivo	O
after	O
injection	O
of	O
2	O
,	O
3	O
,	O
7	O
,	O
8-tetrachlorodibenzo-p-dioxin	O
,	O
a	O
potent	O
ligand	O
of	O
the	O
AHR	B-protein
.	O

The	O
AHR	B-protein
mediates	O
the	O
IL-2	O
induction	O
as	O
shown	O
with	O
AHR	B-protein
-deficient	O
mice	O
.	O

However	O
,	O
in	O
spleen	B-cell_type
cells	I-cell_type
in	O
vitro	O
costimulation	O
via	O
the	O
TCR	B-protein
is	O
necessary	O
for	O
optimal	O
IL-2	B-DNA
gene	I-DNA
induction	O
.	O

Thus	O
,	O
the	O
IL-2	B-DNA
promoter	I-DNA
region	I-DNA
contains	O
novel	O
distal	B-DNA
regulatory	I-DNA
elements	I-DNA
that	O
can	O
be	O
addressed	O
by	O
the	O
AHR	B-protein
to	O
induce	O
IL-2	B-DNA
and	O
can	O
cooperate	O
with	O
the	O
proximal	B-DNA
promoter	I-DNA

-DOCSTART-	O

Visualization	O
of	O
Syk-antigen	B-protein
receptor	I-protein
interactions	O
using	O
green	B-protein
fluorescent	I-protein
protein	I-protein
:	O
differential	O
roles	O
for	O
Syk	B-protein
and	O
Lyn	B-protein
in	O
the	O
regulation	O
of	O
receptor	O
capping	O
and	O
internalization	O
.	O

The	O
cross-linking	O
of	O
the	O
B	B-protein
cell	I-protein
Ag	I-protein
receptor	I-protein
(	O
BCR	B-protein
)	O
is	O
coupled	O
to	O
the	O
stimulation	O
of	O
multiple	O
intracellular	O
signal	O
transduction	O
cascades	O
via	O
receptor-associated	O
,	O
protein	B-protein
tyrosine	I-protein
kinases	I-protein
of	O
both	O
the	O
Src	B-protein
and	I-protein
Syk	I-protein
families	I-protein
.	O

To	O
monitor	O
changes	O
in	O
the	O
subcellular	O
distribution	O
of	O
Syk	B-protein
in	O
B	B-cell_type
cells	I-cell_type
responding	O
to	O
BCR	B-protein
cross-linking	O
,	O
we	O
expressed	O
in	O
Syk	B-protein
-deficient	O
DT40	B-cell_line
B	I-cell_line
cells	I-cell_line
a	O
fusion	B-protein
protein	I-protein
consisting	O
of	O
Syk	B-protein
coupled	O
to	O
green	B-protein
fluorescent	I-protein
protein	I-protein
.	O

Treatment	O
of	O
these	O
cells	O
with	O
anti-IgM	B-protein
Abs	I-protein
leads	O
to	O
the	O
recruitment	O
of	O
the	O
kinase	B-protein
from	O
cytoplasmic	O
and	O
nuclear	O
compartments	O
to	O
the	O
site	O
of	O
the	O
cross-linked	B-protein
receptor	I-protein
at	O
the	O
plasma	O
membrane	O
.	O

The	O
Syk-receptor	B-protein
complexes	I-protein
aggregate	O
into	O
membrane	O
patches	O
that	O
redistribute	O
to	O
form	O
a	O
cap	O
at	O
one	O
pole	O
of	O
the	O
cell	O
.	O

Syk	B-protein
is	O
not	O
demonstrably	O
associated	O
with	O
the	O
internalized	B-protein
receptor	I-protein
.	O

Catalytically	O
active	O
Syk	B-protein
promotes	O
and	O
stabilizes	O
the	O
formation	O
of	O
tightly	B-protein
capped	I-protein
BCR	I-protein
complexes	I-protein
at	O
the	O
plasma	O
membrane	O
.	O

Lyn	B-protein
is	O
not	O
required	O
for	O
the	O
recruitment	O
of	O
Syk	B-protein
to	O
the	O
cross-linked	B-protein
receptor	I-protein
,	O
but	O
is	O
required	O
for	O
the	O
internalization	O
of	O
the	O
clustered	O
BCR	B-protein
complexes	I-protein
.	O

In	O
the	O
absence	O
of	O
Lyn	B-protein
,	O
receptor-Syk	B-protein
complexes	I-protein
at	O
the	O
plasma	O
membrane	O
are	O
long	O
lived	O
,	O
and	O
the	O
receptor-mediated	O
activation	O
of	O
the	O
NF-AT	B-protein
transcription	I-protein
factor	I-protein
is	O
enhanced	O
.	O

Thus	O
,	O
Lyn	B-protein
appears	O
to	O
function	O
to	O
negatively	O
regulate	O
aspects	O
of	O
BCR	B-protein
-dependent	O
signaling	O
by	O
stimulating	O
receptor	O
internalization	O
and	O
down-regulation	O
.	O

-DOCSTART-	O

Renal	O
interstitial	O
fibrosis	O
is	O
reduced	O
in	O
angiotensin	B-protein
II	I-protein
type	I-protein
1a	I-protein
receptor	I-protein
-deficient	O
mice	O
.	O

Unilateral	O
ureteral	O
obstruction	O
(	O
UUO	O
)	O
results	O
in	O
tubulointerstitial	O
fibrosis	O
of	O
the	O
affected	O
kidney	O
by	O
stimulating	O
the	O
renin-angiotensin	O
system	O
.	O

This	O
study	O
established	O
a	O
UUO	O
model	O
in	O
angiotensin	B-protein
type	I-protein
1a	I-protein
receptor	I-protein
(	O
AT1a	B-protein
)	O
deficient	O
(	O
mutant	O
)	O
mice	O
to	O
elucidate	O
the	O
role	O
of	O
angiotensin	B-protein
II	I-protein
through	O
AT1a	B-protein
on	O
the	O
fibrosis	O
of	O
the	O
obstructed	O
kidney	O
(	O
OBK	O
)	O
.	O

The	O
relative	O
volume	O
of	O
the	O
tubulointerstitium	O
was	O
measured	O
by	O
an	O
image	O
analyzer	O
;	O
deposition	O
of	O
collagen	B-protein
types	I-protein
III	I-protein
and	I-protein
IV	I-protein
and	O
monocyte	B-cell_type
/	O
macrophage	B-cell_type
infiltration	O
were	O
histologically	O
examined	O
using	O
specific	B-protein
antibodies	I-protein
.	O

Also	O
determined	O
were	O
the	O
mRNA	O
levels	O
of	O
transforming	B-protein
growth	I-protein
factor-beta	I-protein
by	O
Northern	O
blot	O
analysis	O
.	O

Nuclear	B-protein
factor-kappaB	I-protein
activity	O
was	O
assessed	O
by	O
gel	O
shift	O
assay	O
.	O

UUO	O
in	O
wild	O
mice	O
resulted	O
in	O
a	O
marked	O
expansion	O
of	O
relative	O
volume	O
of	O
the	O
tubulointerstitium	O
,	O
together	O
with	O
increased	O
deposition	O
of	O
collagen	B-protein
types	I-protein
III	I-protein
and	I-protein
IV	I-protein
and	O
number	O
of	O
infiltrated	O
monocytes	B-cell_type
/	O
macrophages	B-cell_type
in	O
the	O
interstitium	O
,	O
relative	O
to	O
sham-operated	O
mice	O
.	O

In	O
comparison	O
,	O
these	O
changes	O
were	O
significantly	O
lower	O
in	O
mutant	O
mice	O
with	O
UUO	O
.	O

The	O
mRNA	O
level	O
of	O
transforming	B-protein
growth	I-protein
factor-beta	I-protein
was	O
significantly	O
higher	O
in	O
the	O
OBK	O
of	O
wild	O
mice	O
with	O
UUO	O
compared	O
with	O
sham-operated	O
mice	O
.	O

In	O
contrast	O
,	O
the	O
increase	O
in	O
mRNA	O
level	O
in	O
the	O
OBK	O
of	O
mutant	O
mice	O
was	O
significantly	O
less	O
than	O
in	O
wild	O
mice	O
.	O

Finally	O
,	O
UUO	O
resulted	O
in	O
activation	O
of	O
nuclear	B-protein
factor-kappaB	I-protein
in	O
wild	O
mice	O
but	O
was	O
inhibited	O
in	O
the	O
OBK	O
of	O
mutant	O
mice	O
.	O

The	O
results	O
provide	O
direct	O
evidence	O
that	O
angiotensin	B-protein
II	I-protein
acting	O
via	O
the	O
AT1a	B-protein
plays	O
a	O
pivotal	O
role	O
in	O
the	O
development	O
of	O
tubulointerstitial	O
fibrosis	O
in	O
UUO	O
.	O

-DOCSTART-	O

HLA-DQ	B-protein
tetramers	I-protein
identify	O
epitope-specific	B-cell_type
T	I-cell_type
cells	I-cell_type
in	O
peripheral	O
blood	O
of	O
herpes	O
simplex	O
virus	O
type	O
2-infected	O
individuals	O
:	O
direct	O
detection	O
of	O
immunodominant	B-cell_type
antigen-responsive	I-cell_type
cells	I-cell_type
.	O

Ag-specific	B-cell_type
CD4+	I-cell_type
T	I-cell_type
cells	I-cell_type
are	O
present	O
in	O
peripheral	O
blood	O
in	O
low	O
frequency	O
,	O
where	O
they	O
undergo	O
recruitment	O
and	O
expansion	O
during	O
immune	O
responses	O
and	O
in	O
the	O
pathogenesis	O
of	O
numerous	O
autoimmune	O
diseases	O
.	O

MHC	B-protein
tetramers	I-protein
,	O
which	O
constitute	O
a	O
labeled	B-protein
MHC-peptide	I-protein
ligand	I-protein
suitable	O
for	O
binding	O
to	O
the	O
Ag-specific	B-protein
receptor	I-protein
on	O
T	B-cell_type
cells	I-cell_type
,	O
provide	O
a	O
novel	O
approach	O
for	O
the	O
detection	O
and	O
characterization	O
of	O
such	O
rare	B-cell_type
cells	I-cell_type
.	O

In	O
this	O
study	O
,	O
we	O
utilized	O
this	O
technology	O
to	O
identify	O
HLA	B-protein
DQ	I-protein
-restricted	O
Ag-specific	B-cell_type
T	I-cell_type
cells	I-cell_type
in	O
the	O
peripheral	O
blood	O
of	O
human	O
subjects	O
and	O
to	O
identify	O
immunodominant	O
epitopes	O
associated	O
with	O
viral	O
infection	O
.	O

Peptides	O
representing	O
potential	O
epitope	B-protein
regions	I-protein
of	O
the	O
VP16	B-protein
protein	I-protein
from	O
HSV-2	O
were	O
loaded	O
onto	O
recombinant	B-protein
DQ0602	I-protein
molecules	I-protein
to	O
generate	O
a	O
panel	O
of	O
Ag-specific	B-protein
DQ0602	I-protein
tetramers	I-protein
.	O

VP16	B-protein
Ag	I-protein
-specific	O
DQ-restricted	B-cell_type
T	I-cell_type
cells	I-cell_type
were	O
identified	O
and	O
expanded	O
from	O
the	O
peripheral	O
blood	O
of	O
HSV-2-infected	O
individuals	O
,	O
representing	O
two	O
predominant	O
epitope	O
specificities	O
.	O

Although	O
the	O
VP16	O
369-380	O
peptide	O
has	O
a	O
lower	O
binding	O
affinity	O
for	O
DQ0602	B-protein
molecules	I-protein
than	O
the	O
VP16	O
33-52	O
peptide	O
,	O
T	B-cell_type
cells	I-cell_type
that	O
recognized	O
the	O
VP16	O
369-380	O
peptide	O
occurred	O
at	O
a	O
much	O
higher	O
frequency	O
than	O
those	O
that	O
were	O
specific	O
for	O
the	O
VP16	O
33-52	O
peptide	O
.	O

-DOCSTART-	O

Down-regulation	O
of	O
BOB.1/OBF.1	B-protein
and	O
Oct2	B-protein
in	O
classical	O
Hodgkin	O
disease	O
but	O
not	O
in	O
lymphocyte	O
predominant	O
Hodgkin	O
disease	O
correlates	O
with	O
immunoglobulin	O
transcription	O
.	O

In	O
contrast	O
to	O
the	O
tumor	B-cell_type
cells	I-cell_type
(	O
L	B-cell_type
&	I-cell_type
H	I-cell_type
cells	I-cell_type
)	O
of	O
lymphocyte	O
predominant	O
Hodgkin	O
disease	O
(	O
LPHD	O
)	O
,	O
Hodgkin	B-cell_type
and	I-cell_type
Reed-Sternberg	I-cell_type
(	I-cell_type
HRS	I-cell_type
)	I-cell_type
cells	I-cell_type
of	O
classical	O
Hodgkin	O
disease	O
(	O
cHD	O
)	O
are	O
unable	O
to	O
transcribe	O
immunoglobulin	B-protein
,	O
despite	O
the	O
presence	O
of	O
rearranged	B-DNA
immunoglobulin	I-DNA
genes	I-DNA
.	O

Although	O
initial	O
studies	O
have	O
suggested	O
crippling	O
immunoglobulin	O
gene	O
mutations	O
to	O
be	O
the	O
cause	O
of	O
absent	O
immunoglobulin	O
expression	O
in	O
cHD	O
,	O
recent	O
work	O
of	O
our	O
group	O
has	O
demonstrated	O
an	O
impaired	O
activation	O
of	O
the	O
immunoglobulin	B-DNA
promoter	I-DNA
as	O
a	O
superior	O
mechanism	O
.	O

As	O
immunoglobulin	O
transcription	O
is	O
mainly	O
regulated	O
by	O
the	O
B-cell	B-protein
transcription	I-protein
factors	I-protein
Oct2	B-protein
and	O
BOB.1/OBF.1	B-protein
,	O
we	O
analyzed	O
35	O
cases	O
of	O
LPHD	O
,	O
32	O
cases	O
of	O
cHD	O
,	O
and	O
2	O
Hodgkin	B-cell_line
disease	I-cell_line
cell	I-cell_line
lines	I-cell_line
for	O
the	O
expression	O
of	O
these	O
transcription	B-protein
factors	I-protein
and	O
also	O
in	O
parallel	O
for	O
immunoglobulin	O
expression	O
.	O

Our	O
results	O
demonstrate	O
an	O
absence	O
of	O
Oct2	B-protein
and/or	O
BOB.1/OBF.1	B-protein
in	O
cHD	O
and	O
a	O
striking	O
overexpression	O
of	O
Oct2	B-protein
in	O
LPHD	O
.	O

Immunoglobulin	O
expression	O
was	O
lacking	O
in	O
cHD	O
but	O
present	O
in	O
LPHD	O
.	O

Furthermore	O
,	O
the	O
reintroduction	O
of	O
BOB.1/OBF.1	B-protein
and	O
Oct2	B-protein
into	O
cultured	O
HRS	B-cell_type
cells	I-cell_type
restored	O
the	O
activity	O
of	O
cotransduced	O
immunoglobulin	B-DNA
promoter	I-DNA
constructs	I-DNA
.	O

Our	O
findings	O
dismiss	O
the	O
concept	O
that	O
the	O
different	O
immunoglobulin	O
expression	O
in	O
cHD	O
and	O
LPHD	O
is	O
due	O
to	O
disrupting	O
mutations	O
of	O
immunoglobulin	B-DNA
V	I-DNA
genes	I-DNA
in	O
cHD	O
but	O
is	O
most	O
likely	O
due	O
to	O
a	O
down-regulation	O
of	O
Oct2	B-protein
and/or	O
BOB.1/OBF.1	B-protein
.	O

This	O
study	O
further	O
revealed	O
Oct2	B-protein
as	O
a	O
new	O
and	O
valuable	O
marker	O
for	O
the	O
identification	O
of	O
L	B-cell_type
&	I-cell_type
H	I-cell_type
cells	I-cell_type
and	O
their	O
distinction	O
from	O
HRS	B-cell_type
cells	I-cell_type
.	O

The	O
impairment	O
of	O
immunoglobulin	O
transcription	O
with	O
a	O
down-regulated	O
synthesis	O
of	O
Oct2	B-protein
and	O
BOB.1/OBF.1	B-protein
is	O
the	O
first	O
established	O
general	O
recurrent	O
defect	O
found	O
in	O
HRS	B-cell_type
cells	I-cell_type
.	O

-DOCSTART-	O

Constitutive	O
expression	O
of	O
NF-kappa	B-protein
B	I-protein
is	O
a	O
characteristic	O
feature	O
of	O
mycosis	O
fungoides	O
:	O
implications	O
for	O
apoptosis	O
resistance	O
and	O
pathogenesis	O
.	O

The	O
NF-kappa	B-protein
B	I-protein
family	I-protein
of	I-protein
transcription	I-protein
factors	I-protein
is	O
an	O
important	O
regulator	O
of	O
genes	O
expressed	O
during	O
inflammatory	O
responses	O
,	O
immunoglobulin	O
(	O
Ig	O
)	O
class	O
switching	O
,	O
cellular	O
differentiation	O
,	O
and	O
apoptosis	O
.	O

Recently	O
,	O
members	O
of	O
the	O
NF-kappaB	B-protein
family	I-protein
,	O
including	O
p65	B-protein
(	I-protein
Rel	I-protein
A	I-protein
)	I-protein
,	O
have	O
been	O
implicated	O
in	O
promoting	O
survival	O
of	O
various	O
hematopoeitic	O
neoplasms	O
,	O
including	O
T	O
cell	O
malignancies	O
such	O
as	O
adult	O
T	O
cell	O
leukemia-lymphoma	O
.	O

We	O
investigated	O
the	O
expression	O
of	O
active	B-protein
NF-kappa	I-protein
B	I-protein
p65	I-protein
(	I-protein
Rel	I-protein
A	I-protein
)	I-protein
in	O
cases	O
of	O
mycosis	O
fungoides	O
(	O
MF	O
)	O
and	O
the	O
effect	O
of	O
chemical	O
inhibitors	O
of	O
NF-kappa	B-protein
B	I-protein
on	O
apoptosis	O
in	O
cutaneous	B-cell_line
T	I-cell_line
cell	I-cell_line
lymphoma	I-cell_line
(	I-cell_line
CTCL	I-cell_line
)	I-cell_line
cell	I-cell_line
lines	I-cell_line
.	O

Paraffin-embedded	O
tissues	O
from	O
23	O
cutaneous	O
lesions	O
and	O
a	O
single	O
lymph	O
node	O
biopsy	O
from	O
patients	O
diagnosed	O
with	O
MF	O
were	O
evaluated	O
for	O
p65	O
(	O
Rel	O
A	O
)	O
expression	O
by	O
using	O
a	O
monoclonal	B-protein
mouse	I-protein
antibody	I-protein
that	O
detects	O
the	O
activated	O
form	O
of	O
p65	B-protein
(	I-protein
Rel	I-protein
A	I-protein
)	I-protein
.	O

Apoptosis	O
after	O
treatment	O
with	O
the	O
NF-kappa	O
B	O
inhibitors	O
gliotoxin	O
,	O
MG132	O
,	O
BAY	O
11-7082	O
,	O
and	O
BAY	O
11-7085	O
was	O
quantitatively	O
measured	O
in	O
the	O
CTCL	B-cell_line
cell	I-cell_line
lines	I-cell_line
HuT-78	I-cell_line
and	I-cell_line
HH	I-cell_line
by	O
propidium	O
iodide	O
(	O
PI	O
)	O
/cell	O
cycle	O
analysis	O
for	O
detection	O
of	O
a	O
hypodiploid	O
(	O
sub-G	O
(	O
0	O
)	O
)	O
population	O
and	O
by	O
determination	O
of	O
increased	O
Annexin	O
V/7-amino-actinomycin	O
D	O
(	O
7-AAD	O
)	O
expression	O
.	O

Nuclear	O
extracts	O
from	O
CTCL	B-cell_type
cells	I-cell_type
before	O
and	O
after	O
chemical	O
inhibition	O
were	O
analyzed	O
for	O
NF-kappa	O
B	O
nuclear	O
DNA-binding	O
activity	O
by	O
electrophoretic	O
mobility	O
shift	O
assay	O
(	O
EMSA	O
)	O
with	O
quantitative	O
densitometry	O
.	O

Nuclear	O
expression	O
of	O
p65	B-protein
(	I-protein
Rel	I-protein
A	I-protein
)	I-protein
before	O
and	O
after	O
treatment	O
with	O
the	O
various	O
inhibitory	O
compounds	O
was	O
measured	O
by	O
immunofluorescence	O
staining	O
in	O
each	O
CTCL	B-cell_line
cell	I-cell_line
line	I-cell_line
.	O

Neoplastic	B-cell_type
T	I-cell_type
lymphocytes	I-cell_type
from	O
22	O
of	O
24	O
cases	O
of	O
MF	O
showed	O
strong	O
nuclear	O
and	O
cytoplasmic	O
expression	O
of	O
active	B-protein
p65	I-protein
(	I-protein
Rel	I-protein
A	I-protein
)	I-protein
.	O

Compared	O
with	O
untreated	O
control	B-cell_type
cells	I-cell_type
,	O
a	O
marked	O
increase	O
in	O
apoptosis	O
,	O
a	O
significant	O
decrease	O
in	O
NF-kappa	O
B	O
DNA-binding	O
activity	O
,	O
and	O
a	O
marked	O
decrease	O
in	O
nuclear	O
p65	O
(	O
Rel	O
A	O
)	O
expression	O
were	O
seen	O
in	O
cells	O
from	O
both	O
CTCL	B-cell_line
cell	I-cell_line
lines	I-cell_line
after	O
chemical	O
NF-kappa	O
B	O
inhibition	O
.	O

These	O
data	O
show	O
that	O
the	O
active	O
form	O
of	O
NF-kappa	B-protein
B	I-protein
p65	I-protein
(	I-protein
Rel	I-protein
A	I-protein
)	I-protein
is	O
commonly	O
expressed	O
in	O
neoplastic	B-cell_type
T	I-cell_type
lymphocytes	I-cell_type
in	O
patients	O
with	O
MF	O
.	O

In	O
CTCL	B-cell_line
cell	I-cell_line
lines	I-cell_line
,	O
the	O
significant	O
decrease	O
in	O
nuclear	O
NF-kappa	O
B	O
expression	O
and	O
the	O
marked	O
increase	O
in	O
spontaneous	O
apoptosis	O
caused	O
by	O
chemical	O
NF-kappa	O
B	O
inhibition	O
suggest	O
a	O
critical	O
role	O
for	O
NF-kappa	B-protein
B	I-protein
in	O
the	O
pathogenesis	O
and	O
tumor	O
cell	O
maintenance	O
of	O
CTCLs	O
.	O

HUM	O
PATHOL	O
31	O
:	O
1482-1490	O
.	O

-DOCSTART-	O

Human	O
T-cell	O
leukemia	O
virus	O
type	O
1	O
tax	B-protein
protein	I-protein
activates	O
transcription	O
through	O
AP-1	B-DNA
site	I-DNA
by	O
inducing	O
DNA	O
binding	O
activity	O
in	O
T	B-cell_type
cells	I-cell_type
.	O

Human	O
T-cell	O
leukemia	O
virus	O
type	O
1	O
(	O
HTLV-1	O
)	O
Tax	B-protein
protein	I-protein
induces	O
the	O
expression	O
of	O
various	O
family	O
members	O
of	O
the	O
transcription	B-protein
factor	I-protein
AP-1	I-protein
,	O
such	O
as	O
c-Jun	B-protein
,	O
JunD	B-protein
,	O
c-Fos	B-protein
,	O
and	O
Fra-1	B-protein
,	O
at	O
the	O
level	O
of	O
RNA	O
expression	O
in	O
T	B-cell_type
cells	I-cell_type
.	O

We	O
examined	O
the	O
activity	O
of	O
Tax	B-protein
in	O
transcription	O
through	O
AP-1-binding	B-DNA
sites	I-DNA
(	O
AP-1	B-DNA
site	I-DNA
)	O
in	O
T	B-cell_type
cells	I-cell_type
.	O

Transient	O
transfection	O
studies	O
showed	O
that	O
Tax	B-protein
activated	O
the	O
expression	O
of	O
a	O
luciferase	B-DNA
gene	I-DNA
regulated	O
by	O
two	O
copies	O
of	O
an	O
AP-1	B-DNA
site	I-DNA
in	O
the	O
human	B-cell_line
Jurkat	I-cell_line
T-cell	I-cell_line
line	I-cell_line
.	O

Tax	B-protein
activates	O
the	O
expression	O
of	O
viral	B-DNA
and	I-DNA
cellular	I-DNA
genes	I-DNA
through	O
two	O
different	O
enhancers	O
:	O
a	O
cAMP-responsive	B-DNA
(	I-DNA
CRE	I-DNA
)	I-DNA
-like	I-DNA
element	I-DNA
and	O
a	O
kappaB	B-DNA
element	I-DNA
.	O

Two	O
Tax	B-protein
mutants	I-protein
differentially	O
activated	O
expression	O
of	O
these	O
two	O
elements	O
.	O

Tax703	B-protein
preferentially	O
activated	O
the	O
kappaB	B-DNA
element	I-DNA
but	O
not	O
the	O
CRE-like	O
one	O
,	O
whereas	O
TaxM22	B-protein
showed	O
the	O
reverse	O
.	O

In	O
addition	O
,	O
Tax703	B-protein
and	O
Tax	B-protein
,	O
but	O
not	O
TaxM22	B-protein
,	O
converted	O
cell	O
growth	O
of	O
a	O
mouse	B-cell_line
T-cell	I-cell_line
line	I-cell_line
from	O
being	O
interleukin	O
(	O
IL	O
)	O
-2-dependent	O
to	O
being	O
IL-2-independent	O
.	O

Unlike	O
the	O
wild-type	B-protein
Tax	I-protein
,	O
Tax703	B-protein
and	O
TaxM22	B-protein
only	O
weakly	O
activated	O
the	O
AP-1	B-DNA
site	I-DNA
in	O
the	O
T-cell	B-cell_line
line	I-cell_line
.	O

Thus	O
,	O
Tax	B-protein
seems	O
to	O
activate	O
the	O
AP-1	B-DNA
site	I-DNA
via	O
mechanisms	O
distinct	O
from	O
those	O
of	O
kappaB	B-DNA
or	I-DNA
CRE-like	I-DNA
elements	I-DNA
,	O
and	O
the	O
activation	O
of	O
the	O
AP-1	B-DNA
site	I-DNA
is	O
dispensable	O
for	O
IL-2-independent	O
growth	O
of	O
CTLL-2	O
.	O

Electrophoretic	O
mobility	O
shift	O
assays	O
showed	O
that	O
Tax	B-protein
induced	O
strong	O
binding	O
activity	O
to	O
an	O
AP-1	B-DNA
site	I-DNA
in	O
CTLL-2	O
,	O
whereas	O
Tax703	B-protein
did	O
not	O
,	O
indicating	O
that	O
the	O
induction	O
of	O
binding	O
activity	O
to	O
the	O
AP-1	B-DNA
site	I-DNA
is	O
essential	O
for	O
the	O
transcriptional	O
activation	O
by	O
Tax	B-protein
.	O

The	O
binding	B-protein
complex	I-protein
induced	O
by	O
Tax	B-protein
in	O
CTLL-2	O
contained	O
JunD	B-protein
and	O
Fra-2	B-protein
.	O

Other	O
AP-1	B-protein
proteins	I-protein
were	O
undetectable	O
.	O

Activation	O
of	O
transcription	O
through	O
the	O
AP-1	B-DNA
site	I-DNA
in	O
Jurkat	B-cell_line
cells	I-cell_line
by	O
JunD	B-protein
and/or	O
Fra-2	B-protein
was	O
weak	O
.	O

c-Jun	B-protein
,	O
JunB	B-protein
,	O
and	O
c-Fos	B-protein
activation	O
was	O
greater	O
,	O
although	O
the	O
level	O
was	O
still	O
less	O
than	O
that	O
with	O
Tax	B-protein
.	O

Thus	O
,	O
the	O
induction	O
of	O
AP-1	B-RNA
mRNA	I-RNA
by	O
Tax	B-protein
may	O
not	O
be	O
sufficient	O
for	O
a	O
complete	O
activation	O
of	O
AP-1	B-DNA
site	I-DNA
by	O
Tax	B-protein
.	O

Our	O
results	O
suggest	O
that	O
Tax	B-protein
activates	O
the	O
transcription	O
of	O
cellular	B-DNA
genes	I-DNA
with	O
AP-1	B-DNA
sites	I-DNA
by	O
inducing	O
the	O
DNA-binding	O
activity	O
of	O
AP-1	B-protein
proteins	I-protein
in	O
T	B-cell_type
cells	I-cell_type
,	O
a	O
mechanism	O
distinct	O
from	O
those	O
of	O
CRE-like	B-DNA
and	I-DNA
kappaB	I-DNA
elements	I-DNA
.	O

Copyright	O
2001	O
Academic	O
Press	O
.	O

-DOCSTART-	O

Strict	O
control	O
of	O
human	O
immunodeficiency	O
virus	O
type	O
1	O
replication	O
by	O
a	O
genetic	O
switch	O
:	O
Tet	O
for	O
Tat	O
.	O

Live-attenuated	O
human	O
immunodeficiency	O
virus	O
type	O
1	O
(	O
HIV-1	O
)	O
variants	O
have	O
shown	O
great	O
promise	O
as	O
AIDS	O
vaccines	O
,	O
but	O
continued	O
replication	O
can	O
lead	O
to	O
the	O
selection	O
of	O
faster-replicating	O
variants	O
that	O
are	O
pathogenic	O
.	O

We	O
therefore	O
designed	O
HIV-1	B-DNA
genomes	I-DNA
that	O
replicate	O
exclusively	O
upon	O
addition	O
of	O
the	O
nontoxic	O
effector	O
doxycycline	O
(	O
dox	O
)	O
.	O

This	O
was	O
achieved	O
by	O
replacement	O
of	O
the	O
viral	O
TAR-Tat	O
system	O
for	O
transcriptional	O
activation	O
by	O
the	O
Escherichia	O
coli-derived	O
Tet	O
system	O
for	O
inducible	O
gene	O
expression	O
.	O

These	O
designer	O
``	O
HIV-rtTA	O
''	O
viruses	O
replicate	O
in	O
a	O
strictly	O
dox-dependent	O
manner	O
both	O
in	O
a	O
T-cell	B-cell_line
line	I-cell_line
and	O
in	O
primary	B-cell_type
blood	I-cell_type
cells	I-cell_type
,	O
and	O
the	O
rate	O
of	O
replication	O
can	O
be	O
fine-tuned	O
by	O
simple	O
variation	O
of	O
the	O
dox	O
concentration	O
.	O

These	O
HIV-rtTA	O
viruses	O
provide	O
a	O
tool	O
to	O
perform	O
genetics	O
,	O
e.g.	O
,	O
selection	O
and	O
optimization	O
experiments	O
,	O
with	O
the	O
E.	O
coli-derived	O
Tet	O
reagents	O
in	O
a	O
eukaryotic	O
background	O
.	O

Furthermore	O
,	O
such	O
viruses	O
may	O
represent	O
improved	O
vaccine	O
candidates	O
because	O
their	O
replication	O
can	O
be	O
turned	O
on	O
and	O
off	O
at	O
will	O
.	O

-DOCSTART-	O

Synovial	O
fluid	O
induced	O
nuclear	B-protein
factor-kappaB	I-protein
DNA	O
binding	O
in	O
a	O
monocytic	B-cell_line
cell	I-cell_line
line	I-cell_line
.	O

OBJECTIVE	O
:	O
To	O
determine	O
the	O
effects	O
of	O
synovial	O
fluids	O
(	O
SF	O
)	O
on	O
DNA	O
binding	O
activity	O
of	O
transcription	B-protein
factor	I-protein
nuclear	B-protein
factor-kappaB	I-protein
(	O
NF-kappaB	B-protein
)	O
in	O
the	O
Mono	B-cell_line
Mac	I-cell_line
6	I-cell_line
monocytic/macrophage	I-cell_line
cell	I-cell_line
line	I-cell_line
as	O
a	O
model	O
for	O
the	O
interaction	O
between	O
SF	O
and	O
synovial	B-cell_type
tissue	I-cell_type
macrophages	I-cell_type
in	O
arthritis	O
.	O

METHODS	O
:	O
Mono	B-cell_line
Mac	I-cell_line
6	I-cell_line
cells	I-cell_line
were	O
incubated	O
with	O
SF	O
from	O
the	O
knee	O
joints	O
of	O
human	O
subjects	O
with	O
rheumatoid	O
arthritis	O
(	O
RA	O
)	O
,	O
undifferentiated	O
seronegative	O
oligoarthritis	O
,	O
and	O
osteoarthritis	O
(	O
OA	O
)	O
.	O

Nuclear	O
extracts	O
prepared	O
from	O
the	O
Mono	B-cell_line
Mac	I-cell_line
6	I-cell_line
cells	I-cell_line
and	O
RA	O
synovial	O
tissue	O
were	O
analyzed	O
by	O
electrophoretic	O
mobility	O
shift	O
analysis	O
(	O
EMSA	O
)	O
for	O
NF-kappaB	B-protein
DNA	I-protein
binding	I-protein
proteins	I-protein
.	O

RESULTS	O
:	O
Induction	O
of	O
NF-kappaB	B-protein
DNA	O
binding	O
by	O
the	O
p65	B-protein
(	I-protein
RelA	I-protein
)	I-protein
/p50	I-protein
heterodimer	I-protein
was	O
observed	O
in	O
response	O
to	O
incubation	O
of	O
Mono	B-cell_line
Mac	I-cell_line
6	I-cell_line
cells	I-cell_line
with	O
SF	O
(	O
20	O
%	O
in	O
culture	O
medium	O
)	O
from	O
5	O
of	O
8	O
subjects	O
with	O
RA	O
,	O
4	O
of	O
5	O
with	O
OA	O
,	O
and	O
none	O
of	O
3	O
with	O
undifferentiated	O
seronegative	O
oligoarthritis	O
.	O

Incubation	O
of	O
SF	O
with	O
neutralizing	B-protein
antibodies	I-protein
against	O
tumor	B-protein
necrosis	I-protein
factor-alpha	I-protein
(	I-protein
TNF-alpha	I-protein
)	I-protein
,	O
but	O
not	O
antibodies	B-protein
against	O
interleukin	B-protein
6	I-protein
(	I-protein
IL-6	I-protein
)	I-protein
,	O
significantly	O
reduced	O
the	O
induction	O
of	O
p65/p50	O
binding	O
activity	O
in	O
SF	O
from	O
subjects	O
with	O
RA	O
and	O
OA	O
.	O

Unexpectedly	O
,	O
a	O
slowly	O
migrating	O
SF	O
inducible	O
NF-kappaB	B-protein
-binding	O
complex	O
was	O
observed	O
in	O
EMSA	O
of	O
Mono	B-cell_line
Mac	I-cell_line
6	I-cell_line
cells	I-cell_line
after	O
incubation	O
with	O
SF	O
from	O
5	O
of	O
8	O
RA	O
and	O
2	O
of	O
5	O
OA	O
subjects	O
.	O

The	O
induction	O
of	O
this	O
complex	O
by	O
SF	O
was	O
not	O
affected	O
by	O
neutralization	O
of	O
TNF-alpha	B-protein
or	O
IL-6	B-protein
in	O
SF	O
,	O
and	O
the	O
complex	O
was	O
not	O
inducible	O
by	O
TNF-alpha	B-protein
,	O
IL-1beta	O
,	O
TNF-alpha	B-protein
/	O
IL-1beta	B-protein
,	O
IL-6	B-protein
,	O
platelet	B-protein
derived	I-protein
growth	I-protein
factor	I-protein
,	O
lipopolysaccharide	O
,	O
or	O
tetradecanoyl	O
phorbol	O
acetate	O
.	O

The	O
slowly	O
migrating	B-protein
complex	I-protein
could	O
not	O
be	O
supershifted	O
with	O
antibodies	B-protein
against	O
NF-kappaB	B-protein
,	O
Jun	B-protein
,	O
or	O
the	O
transcriptional	O
coactivators	O
p300	B-protein
or	O
CBP	B-protein
.	O

A	O
NF-kappaB-binding	B-protein
complex	I-protein
with	O
similar	O
slow	O
mobility	O
was	O
observed	O
in	O
nuclear	O
extracts	O
prepared	O
from	O
fresh	O
human	O
RA	O
synovial	O
tissue	O
.	O

CONCLUSION	O
:	O
Biological	O
activity	O
of	O
TNF-alpha	B-protein
in	O
SF	O
from	O
RA	O
and	O
OA	O
subjects	O
is	O
capable	O
of	O
inducing	O
p65/p50	B-protein
NF-kappaB	I-protein
DNA	O
binding	O
activity	O
in	O
macrophages	B-cell_type
.	O

A	O
property	O
of	O
SF	O
that	O
is	O
independent	O
of	O
TNF-alpha	B-protein
and	O
other	O
cytokines	B-protein
is	O
responsible	O
for	O
the	O
induction	O
of	O
a	O
novel	O
slowly	O
migrating	O
NF-kappaB-binding	B-protein
complex	I-protein
.	O

Soluble	O
mediators	O
in	O
SF	O
of	O
subjects	O
with	O
RA	O
and	O
OA	O
can	O
therefore	O
modulate	O
binding	O
of	O
nuclear	B-protein
proteins	I-protein
to	O
the	O
NF-kappaB	B-protein
binding	I-protein
site	I-protein
in	O
macrophages	B-cell_type
and	O
may	O
play	O
a	O
role	O
in	O
inflammatory	O
gene	O
expression	O
in	O
arthritis	O
.	O

-DOCSTART-	O

Peroxisome	B-protein
proliferator	I-protein
activator	I-protein
receptor-gamma	I-protein
agonists	O
and	O
15-deoxy-Delta	O
(	O
12	O
,	O
14	O
)	O
(	O
12	O
,	O
14	O
)	O
-PGJ	O
(	O
2	O
)	O
induce	O
apoptosis	O
in	O
normal	O
and	O
malignant	O
B-lineage	B-cell_type
cells	I-cell_type
.	O

The	O
research	O
described	O
herein	O
evaluates	O
the	O
expression	O
and	O
functional	O
significance	O
of	O
peroxisome	B-protein
proliferator	I-protein
activator	I-protein
receptor-gamma	I-protein
(	I-protein
PPAR-gamma	I-protein
)	I-protein
on	O
B-lineage	B-cell_type
cells	I-cell_type
.	O

Normal	B-cell_type
mouse	I-cell_type
B	I-cell_type
cells	I-cell_type
and	O
a	O
variety	O
of	O
B	B-cell_type
lymphoma	I-cell_type
cells	I-cell_type
reflective	O
of	O
stages	O
of	O
B	O
cell	O
differentiation	O
(	O
e.g.	O
,	O
70Z/3	O
,	O
CH31	O
,	O
WEHI-231	O
,	O
CH12	O
,	O
and	O
J558	O
)	O
express	O
PPAR-gamma	B-RNA
mRNA	I-RNA
and	O
,	O
by	O
Western	O
blot	O
analysis	O
,	O
the	O
67-kDa	B-protein
PPAR-gamma	I-protein
protein	I-protein
.	O

15-Deoxy-Delta	O
(	O
12	O
,	O
14	O
)	O
-PGJ	O
(	O
2	O
)	O
(	O
15d-PGJ	O
(	O
2	O
)	O
)	O
,	O
a	O
PPAR-gamma	O
agonist	O
,	O
has	O
a	O
dose-dependent	O
antiproliferative	O
and	O
cytotoxic	O
effect	O
on	O
normal	B-cell_type
and	I-cell_type
malignant	I-cell_type
B	I-cell_type
cells	I-cell_type
as	O
shown	O
by	O
[	O
(	O
3	O
)	O
H	O
]	O
thymidine	O
and	O
3-	O
[	O
4	O
,	O
5-dimethylthiazol-2-yl	O
]	O
-2	O
,	O
5-diphenyltetrazolium	O
bromide	O
assays	O
.	O

Only	O
PPAR-gamma	O
agonists	O
(	O
thiazolidinediones	O
)	O
,	O
and	O
not	O
PPAR-alpha	O
agonists	O
,	O
mimicked	O
the	O
effect	O
of	O
15d-PGJ	O
(	O
2	O
)	O
on	O
B-lineage	B-cell_type
cells	I-cell_type
,	O
indicating	O
that	O
the	O
mechanism	O
by	O
which	O
15d-PGJ	O
(	O
2	O
)	O
negatively	O
affects	O
B-lineage	B-cell_type
cells	I-cell_type
involves	O
in	O
part	O
PPAR-gamma	B-protein
.	O

The	O
mechanism	O
by	O
which	O
PPAR-gamma	O
agonists	O
induce	O
cytotoxicity	O
is	O
via	O
apoptosis	O
,	O
as	O
shown	O
by	O
annexin	O
V	O
staining	O
and	O
as	O
confirmed	O
by	O
DNA	O
fragmentation	O
detected	O
using	O
the	O
TUNEL	O
assay	O
.	O

Interestingly	O
,	O
addition	O
of	O
PGF	O
(	O
2alpha	O
)	O
,	O
which	O
was	O
not	O
known	O
to	O
affect	O
lymphocytes	B-cell_type
,	O
dramatically	O
attenuated	O
the	O
deleterious	O
effects	O
of	O
PPAR-gamma	O
agonists	O
on	O
B	B-cell_type
lymphomas	I-cell_type
.	O

Surprisingly	O
,	O
15d-PGJ	O
(	O
2	O
)	O
induced	O
a	O
massive	O
increase	O
in	O
nuclear	O
mitogen-activated	O
protein	O
kinase	O
activation	O
,	O
and	O
pretreatment	O
with	O
PGF	O
(	O
2alpha	O
)	O
blunted	O
the	O
mitogen-activated	O
protein	O
kinase	O
activation	O
.	O

This	O
is	O
the	O
first	O
study	O
evaluating	O
PPAR-gamma	O
expression	O
and	O
its	O
significance	O
on	O
B	B-cell_type
lymphocytes	I-cell_type
.	O

PPAR-gamma	O
agonists	O
may	O
serve	O
as	O
a	O
counterbalance	O
to	O
the	O
stimulating	O
effects	O
of	O
other	O
PGs	O
,	O
namely	O
PGE	O
(	O
2	O
)	O
,	O
which	O
promotes	O
B	O
cell	O
differentiation	O
.	O

Finally	O
,	O
the	O
use	O
of	O
PGs	O
,	O
such	O
as	O
15d-PGJ	O
(	O
2	O
)	O
,	O
and	O
synthetic	O
PPAR-gamma	O
agonists	O
to	O
induce	O
apoptosis	O
in	O
B-lineage	B-cell_type
cells	I-cell_type
may	O
lead	O
to	O
the	O
development	O
of	O
novel	O
therapies	O
for	O
fatal	B-cell_type
B	I-cell_type
lymphomas	I-cell_type

-DOCSTART-	O

Inhibition	O
of	O
STAT3	O
signaling	O
leads	O
to	O
apoptosis	O
of	O
leukemic	B-cell_type
large	I-cell_type
granular	I-cell_type
lymphocytes	I-cell_type
and	O
decreased	O
Mcl-1	O
expression	O
.	O

Large	O
granular	O
lymphocyte	O
(	O
LGL	O
)	O
leukemia	O
is	O
characterized	O
by	O
the	O
expansion	O
of	O
antigen-activated	B-cell_type
cytotoxic	I-cell_type
T	I-cell_type
lymphocytes	I-cell_type
.	O

These	O
leukemic	B-cell_type
cells	I-cell_type
are	O
resistant	O
to	O
Fas-mediated	O
apoptosis	O
despite	O
expressing	O
high	O
levels	O
of	O
Fas	B-protein
.	O

We	O
found	O
that	O
leukemic	B-cell_type
LGL	I-cell_type
from	O
19	O
patients	O
displayed	O
high	O
levels	O
of	O
activated	B-protein
STAT3	I-protein
.	O

Treatment	O
of	O
leukemic	B-cell_type
LGL	I-cell_type
with	O
the	O
JAK-selective	O
tyrosine	O
kinase	O
inhibitor	O
AG-490	O
induced	O
apoptosis	O
with	O
a	O
corresponding	O
decrease	O
in	O
STAT	B-protein
-DNA	O
binding	O
activity	O
.	O

Moreover	O
,	O
using	O
an	O
antisense	O
oligonucleotide	O
approach	O
to	O
diminish	O
STAT3	O
expression	O
,	O
we	O
found	O
that	O
Fas	O
sensitivity	O
was	O
restored	O
in	O
leukemic	B-cell_type
LGL	I-cell_type
.	O

AG-490-induced	O
apoptosis	O
in	O
leukemic	B-cell_type
LGL	I-cell_type
was	O
independent	O
of	O
Bcl-xL	O
or	O
Bcl-2	O
expression	O
.	O

However	O
,	O
we	O
found	O
that	O
the	O
Bcl-2-family	B-protein
protein	I-protein
Mcl-1	I-protein
was	O
significantly	O
reduced	O
by	O
AG-490	O
treatment	O
.	O

Activated	B-protein
STAT3	I-protein
was	O
shown	O
to	O
bind	O
an	O
SIE-related	B-DNA
element	I-DNA
in	O
the	O
murine	B-DNA
mcl-1	I-DNA
promoter	I-DNA
.	O

Using	O
a	O
luciferase	O
reporter	O
assay	O
,	O
we	O
demonstrated	O
that	O
v-src	O
overexpression	O
in	O
NIH3T3	O
induced	O
STAT3	B-protein
-dependent	O
transcriptional	O
activity	O
from	O
the	O
mcl-1	B-DNA
promoter	I-DNA
and	O
increased	O
endogenous	O
Mcl-1	O
protein	O
levels	O
.	O

We	O
conclude	O
that	O
STAT3	B-protein
activation	O
contributed	O
to	O
accumulation	O
of	O
the	O
leukemic	B-cell_type
LGL	I-cell_type
clones	I-cell_type
.	O

These	O
findings	O
suggest	O
that	O
investigation	O
should	O
focus	O
on	O
novel	O
strategies	O
targeting	O
STAT3	B-protein
in	O
the	O
treatment	O
of	O
LGL	O
leukemia	O
.	O

-DOCSTART-	O

Decreased	O
immediate	O
inflammatory	B-DNA
gene	I-DNA
induction	O
in	O
activating	B-protein
transcription	I-protein
factor-2	I-protein
mutant	O
mice	O
.	O

Transcription	B-protein
factor	I-protein
activating	I-protein
transcription	I-protein
factor	I-protein
(	I-protein
ATF	I-protein
)	I-protein
-2	I-protein
is	O
activated	O
by	O
inflammatory	O
signals	O
transduced	O
by	O
the	O
JNK	O
and	O
p38	O
MAP	O
kinase	O
pathways	O
.	O

To	O
better	O
define	O
the	O
role	O
of	O
ATF-2	B-protein
in	O
inflammation	O
,	O
adult	O
mice	O
expressing	O
small	O
amounts	O
of	O
a	O
mutant	B-protein
ATF-2	I-protein
protein	I-protein
were	O
challenged	O
with	O
lipopolysaccharide	O
(	O
LPS	O
)	O
,	O
anti-CD3	B-protein
antibody	I-protein
or	O
virus	O
.	O

Within	O
3	O
h	O
of	O
challenge	O
by	O
LPS	O
,	O
ATF-2	O
mutant	O
mice	O
had	O
decreased	O
induction	O
of	O
the	O
adhesion	B-protein
molecules	I-protein
E-selectin	B-protein
,	O
P-selectin	B-protein
and	O
VCAM-1	B-protein
as	O
well	O
as	O
the	O
cytokines	B-protein
tumor	B-protein
necrosis	I-protein
factor-alpha	I-protein
,	O
IL-1beta	B-protein
and	O
IL-6	B-protein
compared	O
with	O
control	O
mice	O
.	O

Stimulation	O
of	O
T	B-cell_type
lymphocytes	I-cell_type
by	O
anti-CD3	B-protein
antibody	I-protein
also	O
showed	O
less	O
induction	O
of	O
IL-1	B-protein
and	O
IL-6	B-protein
in	O
ATF-2	O
mutant	O
tissues	O
.	O

ATF-2	B-cell_type
mutant	I-cell_type
thymocytes	I-cell_type
treated	O
with	O
anti-CD3	B-protein
antibody	I-protein
in	O
vitro	O
demonstrated	O
reduced	O
induction	O
of	O
c-Jun	B-protein
,	O
JunB	B-protein
,	O
JunD	B-protein
and	O
Fra-2	B-protein
.	O

However	O
,	O
similar	O
to	O
what	O
was	O
observed	O
after	O
p38	O
kinase	O
inhibition	O
in	O
normal	O
mice	O
,	O
relative	O
ATF-2	O
deficiency	O
did	O
not	O
prevent	O
the	O
development	O
of	O
a	O
mononuclear	B-cell_type
cell	I-cell_type
infiltrate	O
in	O
the	O
week	O
following	O
an	O
inflammatory	O
stimulus	O
.	O

ATF-2	O
mutant	O
mice	O
proved	O
more	O
susceptible	O
to	O
death	O
than	O
control	O
mice	O
from	O
LPS	O
plus	O
D-galactosamine	O
injection	O
or	O
Coxsackievirus	O
B3	O
infection	O
and	O
had	O
a	O
higher	O
incidence	O
of	O
mononuclear	O
pulmonary	O
infiltrates	O
after	O
exposure	O
to	O
Herpes	O
simplex	O
virus-1	O
.	O

ATF-2	B-protein
is	O
essential	O
for	O
maximal	O
immediate	O
induction	O
of	O
adhesion	B-protein
molecules	I-protein
and	O
cytokine	B-DNA
genes	I-DNA
,	O
but	O
at	O
later	O
time	O
points	O
may	O
even	O
protect	O
against	O
overactive	O
immune	O
responses	O
.	O

-DOCSTART-	O

LIGHT	B-protein
,	O
a	O
TNF-like	B-protein
molecule	I-protein
,	O
costimulates	O
T	O
cell	O
proliferation	O
and	O
is	O
required	O
for	O
dendritic	B-cell_type
cell	I-cell_type
-mediated	O
allogeneic	O
T	O
cell	O
response	O
.	O

LIGHT	B-protein
is	O
a	O
recently	O
identified	O
member	O
of	O
the	O
TNF	B-protein
superfamily	I-protein
and	O
its	O
receptors	O
,	O
herpesvirus	B-protein
entry	I-protein
mediator	I-protein
and	O
lymphotoxin	B-protein
beta	I-protein
receptor	I-protein
,	O
are	O
found	O
in	O
T	B-cell_type
cells	I-cell_type
and	O
stromal	B-cell_type
cells	I-cell_type
.	O

In	O
this	O
study	O
,	O
we	O
demonstrate	O
that	O
LIGHT	B-protein
is	O
selectively	O
expressed	O
on	O
immature	B-cell_type
dendritic	I-cell_type
cells	I-cell_type
(	O
DCs	B-cell_type
)	O
generated	O
from	O
human	B-cell_type
PBMCs	I-cell_type
.	O

In	O
contrast	O
,	O
LIGHT	B-protein
is	O
not	O
detectable	O
in	O
DCs	B-cell_type
either	O
freshly	O
isolated	O
from	O
PBMCs	B-cell_type
or	O
rendered	O
mature	O
in	O
vitro	O
by	O
LPS	O
treatment	O
.	O

Blockade	O
of	O
LIGHT	B-protein
by	O
its	O
soluble	B-protein
receptors	I-protein
,	O
lymphotoxin	B-protein
beta	I-protein
receptor-Ig	I-protein
or	O
HVEM-Ig	B-protein
,	O
inhibits	O
the	O
induction	O
of	O
DC	B-cell_type
-mediated	O
primary	O
allogeneic	O
T	O
cell	O
response	O
.	O

Furthermore	O
,	O
engagement	O
of	O
LIGHT	B-protein
costimulates	O
human	O
T	O
cell	O
proliferation	O
,	O
amplifies	O
the	O
NF-kappaB	O
signaling	O
pathway	O
,	O
and	O
preferentially	O
induces	O
the	O
production	O
of	O
IFN-gamma	B-protein
,	O
but	O
not	O
IL-4	B-protein
,	O
in	O
the	O
presence	O
of	O
an	O
antigenic	O
signal	O
.	O

Our	O
results	O
suggest	O
that	O
LIGHT	B-protein
is	O
a	O
costimulatory	B-protein
molecule	I-protein
involved	O
in	O
DC	B-cell_type
-mediated	O
cellular	O
immune	O
responses	O
.	O

-DOCSTART-	O

Suppression	O
of	O
HIV	O
type	O
1	O
replication	O
by	O
a	O
dominant-negative	B-protein
Ets-1	I-protein
mutant	I-protein
.	O

Activity	O
of	O
the	O
distal	B-DNA
region	I-DNA
of	O
the	O
human	O
immunodeficiency	O
virus	O
(	O
HIV-1	O
)	O
long	B-DNA
terminal	I-DNA
repeat	I-DNA
(	O
LTR	B-DNA
)	O
,	O
which	O
contains	O
binding	B-DNA
sites	I-DNA
for	O
the	O
Ets-1	B-protein
and	I-protein
USF-1	I-protein
proteins	I-protein
,	O
is	O
integral	O
for	O
HIV-1	O
replication	O
.	O

The	O
Ets-1	B-protein
and	I-protein
USF-1	I-protein
proteins	I-protein
play	O
a	O
critical	O
role	O
in	O
the	O
activity	O
of	O
the	O
HIV-1	B-DNA
LTR	I-DNA
distal	I-DNA
enhancer	I-DNA
region	I-DNA
,	O
as	O
indicated	O
by	O
the	O
potent	O
dominant	O
negative	O
effect	O
of	O
a	O
mutant	B-DNA
Ets-1	I-DNA
lacking	I-DNA
trans-activation	I-DNA
domains	I-DNA
on	O
the	O
transcriptional	O
activity	O
of	O
the	O
LTR	B-DNA
.	O

To	O
determine	O
the	O
biological	O
relevance	O
of	O
the	O
Ets-1	B-protein
and	I-protein
USF-1	I-protein
proteins	I-protein
in	O
HIV-1	O
replication	O
,	O
we	O
examined	O
the	O
effect	O
of	O
expression	O
of	O
the	O
dominant-negative	O
mutant	O
of	O
Ets-1	B-DNA
(	O
dnEts-1	B-DNA
)	O
on	O
HIV-1	O
infection	O
of	O
T	B-cell_type
cells	I-cell_type
.	O

We	O
demonstrated	O
that	O
expression	O
of	O
dnEts	B-DNA
markedly	O
suppressed	O
HIV-1	O
infection	O
of	O
a	O
T	B-cell_line
cell	I-cell_line
line	I-cell_line
.	O

This	O
finding	O
indicates	O
that	O
formation	O
of	O
a	O
transcriptionaly	B-protein
active	I-protein
USF-1/Ets-1	I-protein
complex	I-protein
is	O
important	O
in	O
the	O
productive	O
infection	O
of	O
cells	O
by	O
HIV-1	O
,	O
and	O
suggests	O
that	O
inhibition	O
of	O
the	O
interaction	O
between	O
USF-1	B-protein
and	O
Ets-1	B-protein
with	O
the	O
HIV-1	B-DNA
LTR	I-DNA
may	O
provide	O
a	O
new	O
target	O
for	O
anti-HIV-1	O
gene	O
therapy	O
.	O

-DOCSTART-	O

Human	B-cell_type
eosinophils	I-cell_type
constitutively	O
express	O
nuclear	B-protein
factor	I-protein
of	I-protein
activated	I-protein
T	I-protein
cells	I-protein
p	I-protein
and	I-protein
c	I-protein
.	O

BACKGROUND	O
:	O
Eosinophils	B-cell_type
are	O
now	O
known	O
to	O
produce	O
a	O
variety	O
of	O
proinflammatory	B-protein
cytokines	I-protein
,	O
although	O
the	O
molecular	O
factors	O
that	O
regulate	O
their	O
production	O
are	O
poorly	O
understood	O
.	O

The	O
expression	O
of	O
almost	O
all	O
of	O
the	O
cytokines	B-protein
produced	O
by	O
eosinophils	B-cell_type
,	O
including	O
the	O
proallergic	B-protein
cytokine	I-protein
IL-4	I-protein
,	O
is	O
now	O
known	O
to	O
be	O
regulated	O
at	O
the	O
level	O
of	O
transcription	O
by	O
members	O
of	O
the	O
nuclear	B-protein
factor	I-protein
of	I-protein
activated	I-protein
T	I-protein
cells	I-protein
(	O
NFAT	B-protein
)	O
family	O
of	O
transcription	O
factors	O
.	O

OBJECTIVE	O
:	O
We	O
sought	O
to	O
characterize	O
the	O
expression	O
of	O
different	O
NFAT	B-protein
proteins	I-protein
in	O
resting	B-cell_type
and	I-cell_type
activated	I-cell_type
eosinophils	I-cell_type
.	O

METHODS	O
:	O
Nuclear	O
and	O
whole	O
cell	O
extracts	O
were	O
obtained	O
from	O
both	O
peripheral	B-cell_type
blood	I-cell_type
eosinophils	I-cell_type
and	O
those	O
obtained	O
from	O
bronchoalveolar	O
lavage	O
fluid	O
of	O
asthmatic	O
subjects	O
after	O
endobronchial	O
allergen	O
challenge	O
.	O

NFAT	B-protein
expression	O
was	O
determined	O
by	O
using	O
immunoprecipitation	O
and	O
Western	O
blot	O
analysis	O
,	O
DNA-binding	O
assays	O
,	O
and	O
RT-PCR	O
analysis	O
of	O
eosinophil	B-RNA
mRNA	I-RNA
.	O

RESULTS	O
:	O
Both	O
peripheral	B-cell_type
blood	I-cell_type
and	I-cell_type
bronchoalveolar	I-cell_type
lavage	I-cell_type
fluid	I-cell_type
eosinophils	I-cell_type
expressed	O
NFATp	B-protein
and	I-protein
NFATc	I-protein
protein	I-protein
.	O

Unlike	O
activated	B-cell_type
T	I-cell_type
cells	I-cell_type
,	O
which	O
express	O
multiple	O
NFATc	O
isoforms	O
,	O
eosinophils	B-cell_type
preferentially	O
express	O
the	O
approximately	O
85-kd	O
isoform	O
.	O

In	O
addition	O
,	O
eosinophils	B-cell_type
were	O
found	O
to	O
constitutively	O
express	O
NFATc	B-RNA
mRNA	I-RNA
.	O

A	O
brief	O
incubation	O
with	O
the	O
T	B-protein
(	I-protein
H	I-protein
)	I-protein
2	I-protein
cytokines	I-protein
IL-4	B-protein
and	O
IL-5	B-protein
was	O
sufficient	O
to	O
induce	O
the	O
nuclear	O
translocation	O
of	O
NFATc	B-protein
.	O

Eosinophil	O
nuclear	O
extracts	O
contain	O
multiple	O
factors	O
that	O
can	O
specifically	O
recognize	O
the	O
IL-4	B-DNA
promoter	I-DNA
P1	I-DNA
NFAT	I-DNA
site	I-DNA
in	O
DNA-binding	O
assays	O
,	O
including	O
NFATp	B-protein
.	O

CONCLUSION	O
:	O
NFATp	B-protein
and	O
NFATc	B-protein
can	O
regulate	O
the	O
expression	O
of	O
cytokines	B-protein
and	O
other	O
genes	O
in	O
eosinophils	B-cell_type
but	O
appear	O
to	O
be	O
regulated	O
by	O
a	O
novel	O
signal	O
transduction	O
mechanism	O
in	O
these	O
cells	O
.	O

-DOCSTART-	O

BLyS	B-protein
BINDS	O
TO	O
B	B-cell_type
CELLS	I-cell_type
WITH	O
HIGH	O
AFFINITY	O
AND	O
INDUCES	O
ACTIVATION	O
OF	O
THE	O
TRANSCRIPTION	B-protein
FACTORS	I-protein
NF-kappaB	B-protein
AND	O
ELF-1	B-protein
.	O

B	B-protein
lymphocyte	I-protein
stimulator	I-protein
(	O
BLyS	B-protein
)	O
is	O
a	O
novel	O
member	O
of	O
the	O
TNF	B-protein
family	I-protein
of	I-protein
proteins	I-protein
expressed	O
by	O
myeloid	B-cell_type
cells	I-cell_type
as	O
membrane-bound	O
and	O
soluble	O
forms	O
.	O

BLyS	B-protein
was	O
shown	O
to	O
act	O
specifically	O
on	O
B	B-cell_type
cells	I-cell_type
,	O
inducing	O
proliferation	O
and	O
immunoglobulin	O
production	O
both	O
in	O
vitro	O
and	O
in	O
vivo	O
.	O

The	O
present	O
study	O
was	O
undertaken	O
to	O
characterize	O
binding	O
of	O
radiolabeled	B-protein
BLyS	I-protein
to	O
its	O
cognate	B-protein
receptor	I-protein
on	O
human	B-cell_type
B	I-cell_type
lymphocytes	I-cell_type
and	O
examine	O
intracellular	O
events	O
initiated	O
by	O
BLyS	B-protein
binding	O
.	O

Similar	O
to	O
other	O
TNF	B-protein
family	I-protein
members	I-protein
,	O
BLyS	B-protein
is	O
present	O
in	O
solution	O
as	O
a	O
homotrimer	B-protein
as	O
determined	O
by	O
gel	O
filtration	O
chromatography	O
and	O
light	O
scattering	O
analysis	O
.	O

BLyS	B-protein
binding	O
to	O
B	B-cell_type
cells	I-cell_type
is	O
specific	O
as	O
other	O
TNF	B-protein
family	I-protein
members	I-protein
tested	O
did	O
not	O
compete	O
for	O
(	O
125	O
)	O
I-	O
BLyS	B-protein
binding	O
.	O

Analysis	O
of	O
equilibrium	O
binding	O
of	O
(	B-protein
125	I-protein
)	I-protein
I-labeled	I-protein
BLyS	I-protein
to	O
purified	O
human	B-cell_type
tonsillar	I-cell_type
B	I-cell_type
cells	I-cell_type
demonstrated	O
saturable	O
binding	O
.	O

Scatchard	O
analysis	O
of	O
the	O
binding	O
data	O
revealed	O
a	O
single	O
class	O
of	O
high-affinity	O
binding	O
on	O
human	B-cell_type
B	I-cell_type
cells	I-cell_type
with	O
approximately	O
2600	O
binding	O
sites	O
per	O
cell	O
and	O
an	O
apparent	O
dissociation	O
constant	O
(	O
K	O
(	O
D	O
)	O
)	O
of	O
about	O
0.1	O
nM	O
.	O

In	O
addition	O
we	O
report	O
that	O
BLyS	B-protein
binding	O
to	O
B	B-cell_type
cells	I-cell_type
results	O
in	O
the	O
activation	O
of	O
NF-kappaB	B-protein
and	O
the	O
Ets	B-protein
family	I-protein
transcription	I-protein
factor	I-protein
,	O
ELF-1	B-protein
,	O
and	O
in	O
the	O
induction	O
of	O
mRNA	O
for	O
Polo-like	B-protein
kinase	I-protein
(	O
PLK	B-protein
)	O
.	O

Copyright	O
2001	O
Academic	O
Press	O
.	O

-DOCSTART-	O

Design	O
and	O
use	O
of	O
an	O
inducibly	O
activated	O
human	O
immunodeficiency	O
virus	O
type	O
1	O
Nef	B-protein
to	O
study	O
immune	O
modulation	O
.	O

The	O
Nef	B-protein
protein	I-protein
of	O
the	O
human	O
immunodeficiency	O
virus	O
type	O
1	O
(	O
HIV-1	O
)	O
has	O
been	O
shown	O
to	O
enhance	O
the	O
infectivity	O
of	O
virus	O
particles	O
,	O
downmodulate	O
cell	B-protein
surface	I-protein
proteins	I-protein
,	O
and	O
associate	O
with	O
many	O
intracellular	B-protein
proteins	I-protein
that	O
are	O
thought	O
to	O
facilitate	O
HIV	O
infection	O
.	O

One	O
of	O
the	O
challenges	O
in	O
defining	O
the	O
molecular	O
events	O
regulated	O
by	O
Nef	B-protein
has	O
been	O
obtaining	O
good	O
expression	O
of	O
Nef	B-protein
protein	I-protein
in	O
T	B-cell_type
cells	I-cell_type
.	O

This	O
has	O
been	O
attributed	O
to	O
effects	O
of	O
Nef	B-protein
on	O
cell	O
proliferation	O
and	O
apoptosis	O
.	O

We	O
have	O
designed	O
a	O
Nef	B-protein
protein	I-protein
that	O
is	O
readily	O
expressed	O
in	O
T-cell	B-cell_line
lines	I-cell_line
and	O
whose	O
function	O
is	O
inducibly	O
activated	O
.	O

It	O
is	O
composed	O
of	O
a	O
fusion	O
between	O
full-length	B-protein
Nef	I-protein
and	O
the	O
estrogen	B-protein
receptor	I-protein
hormone-binding	I-protein
domain	I-protein
(	O
Nef-ER	B-protein
)	O
.	O

The	O
Nef-ER	B-protein
is	O
kept	O
in	O
an	O
inactive	O
state	O
due	O
to	O
steric	O
hindrance	O
,	O
and	O
addition	O
of	O
the	O
membrane-permeable	O
drug	O
4-hydroxytamoxifen	O
(	O
4-HT	O
)	O
,	O
which	O
binds	O
to	O
the	O
ER	B-protein
domain	I-protein
,	O
leads	O
to	O
inducible	O
activation	O
of	O
Nef-ER	B-protein
within	O
cells	O
.	O

We	O
demonstrate	O
that	O
Nef-ER	B-protein
inducibly	O
associates	O
with	O
the	O
62-kDa	B-protein
Ser/Thr	I-protein
kinase	I-protein
and	O
is	O
localized	O
to	O
specific	O
membrane	O
microdomains	O
(	O
lipid	O
rafts	O
)	O
only	O
after	O
activation	O
.	O

Using	O
this	O
inducible	O
Nef	B-protein
,	O
we	O
also	O
compared	O
the	O
specific	O
requirements	O
for	O
CD4	B-protein
and	O
HLA-A2	B-protein
downmodulation	O
in	O
a	O
SupT1	B-cell_line
T-cell	I-cell_line
line	I-cell_line
.	O

Half-maximal	O
downmodulation	O
of	O
cell	B-protein
surface	I-protein
CD4	I-protein
required	O
very	O
little	O
active	O
Nef-ER	B-protein
and	O
occurred	O
as	O
early	O
as	O
4	O
h	O
after	O
addition	O
of	O
4-HT	O
.	O

In	O
contrast	O
,	O
50	O
%	O
downmodulation	O
of	O
HLA-A2	B-protein
by	O
Nef	B-protein
required	O
16	O
to	O
24	O
h	O
and	O
about	O
50-	O
to	O
100-fold-greater	O
concentrations	O
of	O
4-HT	O
.	O

These	O
data	O
suggest	O
that	O
HLA-A2	B-protein
downmodulation	O
may	O
require	O
certain	O
threshold	O
levels	O
of	O
active	O
Nef	B-protein
.	O

The	O
differential	O
timing	O
of	O
CD4	B-protein
and	O
HLA-A2	B-protein
downmodulation	O
may	O
have	O
implications	O
for	O
HIV	O
pathogenesis	O
and	O
immune	O
evasion	O
.	O

-DOCSTART-	O

Regulation	O
of	O
chemokine	O
mRNA	O
expression	O
in	O
a	O
rat	O
model	O
of	O
vanadium-induced	O
pulmonary	O
inflammation	O
.	O

Environmental	O
and	O
occupational	O
exposure	O
to	O
vanadium	O
dusts	O
results	O
in	O
toxic	O
effects	O
mainly	O
confined	O
to	O
the	O
respiratory	O
system	O
.	O

Using	O
a	O
rat	O
model	O
of	O
acute	O
lung	O
inflammation	O
induced	O
by	O
intratracheal	O
instillation	O
of	O
sodium	O
metavanadate	O
(	O
NaVO3	O
)	O
at	O
the	O
dose	O
of	O
200	O
microg	O
V/kg	O
,	O
we	O
investigated	O
the	O
relationship	O
between	O
the	O
cytologic	O
characterization	O
of	O
pulmonary	O
inflammation	O
and	O
the	O
expression	O
of	O
chemokine	B-RNA
mRNA	I-RNA
.	O

Significant	O
polymorphonuclear	B-cell_type
leukocyte	I-cell_type
(	O
PMN	B-cell_type
)	O
influx	O
(	O
P	O
<	O
0.01	O
)	O
into	O
the	O
lung	O
was	O
noted	O
4	O
h	O
after	O
NaVO3	O
instillation	O
,	O
whereas	O
alveolar	B-cell_type
macrophages	I-cell_type
(	O
AMs	B-cell_type
)	O
in	O
bronchoalveolar	B-cell_type
lavage	I-cell_type
(	I-cell_type
BAL	I-cell_type
)	I-cell_type
cells	I-cell_type
appeared	O
to	O
decrease	O
significantly	O
.	O

In	O
contrast	O
,	O
neither	O
PMNs	B-cell_type
nor	O
AMs	B-cell_type
changed	O
substantially	O
1	O
h	O
after	O
NaVO3	O
instillation	O
.	O

By	O
Northern	O
analysis	O
,	O
macrophage	B-RNA
inflammatory	I-RNA
protein	I-RNA
(	I-RNA
MIP	I-RNA
)	I-RNA
-2	I-RNA
mRNA	I-RNA
in	O
BAL	B-cell_type
cells	I-cell_type
increased	O
markedly	O
1	O
h	O
after	O
NaVO3	O
instillation	O
and	O
reduced	O
a	O
little	O
bit	O
at	O
4	O
h	O
,	O
whereas	O
MIP-1alpha	B-RNA
mRNA	I-RNA
in	O
BAL	B-cell_type
cells	I-cell_type
was	O
expressed	O
relatively	O
high	O
1	O
h	O
after	O
NaVO3	O
instillation	O
,	O
although	O
a	O
basal	O
expression	O
was	O
detected	O
in	O
control	O
group	O
,	O
and	O
returned	O
rapidly	O
nearly	O
to	O
control	O
level	O
at	O
4	O
h	O
.	O

Since	O
MIP-2	B-protein
is	O
a	O
potent	O
PMN	B-cell_type
chemoattractant	O
and	O
MIP-1alpha	B-protein
is	O
a	O
potent	O
macrophage/monocyte	O
chemoattractant	O
has	O
been	O
well	O
known	O
.	O

The	O
facts	O
that	O
PMN	B-cell_type
influx	O
was	O
preceded	O
by	O
increased	O
MIP-2	O
mRNA	O
expression	O
,	O
suggesting	O
that	O
MIP-2	B-protein
is	O
involved	O
in	O
the	O
development	O
of	O
NaVO3-induced	O
pulmonary	O
inflammation	O
,	O
whereas	O
increased	O
MIP-1alpha	B-RNA
mRNA	I-RNA
expression	O
was	O
followed	O
by	O
decreased	O
AMs	B-cell_type
in	O
BAL	B-cell_type
cells	I-cell_type
,	O
suggesting	O
AMs	B-cell_type
might	O
be	O
activated	O
by	O
MIP-1alpha	B-protein
,	O
adherent	O
to	O
the	O
lining	O
surface	O
of	O
the	O
airways	O
and	O
then	O
resistant	O
to	O
be	O
washed	O
out	O
.	O

To	O
delineate	O
the	O
mechanisms	O
of	O
transcriptional	O
activation	O
,	O
we	O
recently	O
cloned	O
the	O
5'-flanking	B-DNA
region	I-DNA
of	O
the	O
MIP-2	B-DNA
gene	I-DNA
.	O

The	O
promotor	B-DNA
region	I-DNA
contains	O
consensus	B-DNA
binding	I-DNA
sites	I-DNA
for	O
transcription	B-protein
factor	I-protein
nuclear	I-protein
factor	I-protein
kappaB	I-protein
(	O
NF-kappaB	B-protein
)	O
and	O
activator	B-protein
protein-1	I-protein
(	O
AP-1	B-protein
)	O
.	O

Using	O
electrophoretic	O
mobility	O
shift	O
assay	O
,	O
increased	O
nuclear	O
NF-kappaB	B-protein
,	O
not	O
AP-1	B-protein
,	O
binding	O
activity	O
was	O
detected	O
1	O
h	O
after	O
NaVO3	O
instillation	O
,	O
which	O
correlated	O
with	O
the	O
induction	O
of	O
MIP-2	B-RNA
mRNA	I-RNA
.	O

p65	B-protein
(	I-protein
Rel	I-protein
A	I-protein
)	I-protein
and	O
p50	B-protein
protein	I-protein
appears	O
to	O
be	O
involved	O
in	O
MIP-2	O
NF-kappaB	B-protein
binding	O
.	O

Taken	O
together	O
,	O
our	O
studies	O
suggest	O
that	O
MIP-2	B-protein
is	O
an	O
important	O
mediator	O
of	O
NaVO3-induced	O
pulmonary	O
inflammation	O
in	O
the	O
rat	O
model	O
.	O

In	O
addition	O
,	O
elevated	O
MIP-2	B-RNA
mRNA	I-RNA
levels	O
are	O
accompanied	O
by	O
increased	O
NF-kappaB	B-protein
binding	O
activity	O
in	O
BAL	B-cell_type
cells	I-cell_type
,	O
suggesting	O
possible	O
MIP-2	B-protein
transcriptional	O
regulation	O
through	O
NF-kappaB	B-protein
.	O

-DOCSTART-	O

Activation	O
of	O
signal	O
transduction	O
and	O
apoptosis	O
in	O
healthy	B-cell_type
lymphomonocytes	I-cell_type
exposed	O
to	O
bystander	O
HIV-1-infected	B-cell_type
cells	I-cell_type
.	O

Persistent	O
activation	O
of	O
the	O
immune	O
system	O
is	O
one	O
of	O
the	O
hallmarks	O
of	O
HIV-1	O
infection	O
.	O

In	O
this	O
study	O
we	O
analysed	O
the	O
induction	O
of	O
factors	O
involved	O
in	O
cytokine	O
signal	O
transduction	O
,	O
such	O
as	O
STAT	B-protein
1	I-protein
proteins	I-protein
and	O
IRF-1	B-RNA
mRNA	I-RNA
,	O
in	O
normal	B-cell_type
peripheral	I-cell_type
blood	I-cell_type
mononuclear	I-cell_type
cells	I-cell_type
(	O
PBMC	B-cell_type
)	O
exposed	O
to	O
HIV-infected	B-cell_type
cells	I-cell_type
,	O
and	O
the	O
induction	O
of	O
apoptosis	O
.	O

Western	O
blot	O
analyses	O
and	O
reverse	O
transcriptase-polymerase	O
chain	O
reaction	O
results	O
indicate	O
that	O
both	O
cells	O
infected	O
with	O
a	O
X4	O
strain	O
and	O
cells	O
infected	O
with	O
a	O
R5	O
strain	O
are	O
able	O
to	O
increase	O
intracellular	O
levels	O
of	O
STAT	B-protein
1alpha	I-protein
and	I-protein
beta	I-protein
proteins	O
as	O
well	O
as	O
IRF-1	B-RNA
mRNA	I-RNA
.	O

This	O
effect	O
was	O
prevented	O
by	O
neutralizing	O
antibodies	O
against	O
interferon-alpha	B-protein
(	O
IFN-alpha	B-protein
)	O
.	O

HIV-1-infected	B-cell_type
cells	I-cell_type
dose-dependently	O
induced	O
apoptotic	O
commitment	O
in	O
normal	B-cell_type
PBMC	I-cell_type
,	O
as	O
revealed	O
by	O
DNA	O
fragmentation	O
analysis	O
,	O
but	O
this	O
was	O
not	O
accompanied	O
by	O
an	O
increase	O
of	O
caspase-3	O
activity	O
,	O
even	O
if	O
a	O
slight	O
up-regulation	O
of	O
IL-1beta-converting	B-RNA
enzyme	I-RNA
mRNA	I-RNA
was	O
detected	O
.	O

Apoptosis	O
induction	O
could	O
be	O
abrogated	O
mainly	O
by	O
antibodies	B-protein
against	O
tumour	B-protein
necrosis	I-protein
factor-alpha	I-protein
(	O
TNF-alpha	B-protein
)	O
and	O
,	O
to	O
a	O
lesser	O
extent	O
,	O
by	O
antibodies	B-protein
against	O
IFN-gamma	B-protein
.	O

All	O
these	O
findings	O
suggest	O
that	O
uninfected	B-cell_type
PBMC	I-cell_type
can	O
undergo	O
activation	O
of	O
signal	O
transduction	O
and	O
apoptosis	O
after	O
exposure	O
to	O
bystander	O
HIV-infected	B-cell_type
cells	I-cell_type
,	O
subsequent	O
to	O
the	O
induction	O
of	O
cytokines	B-protein
such	O
as	O
IFNs	B-protein
and	O
TNF-alpha	B-protein
.	O

-DOCSTART-	O

The	O
physical	O
association	O
of	O
protein	B-protein
kinase	I-protein
C	I-protein
theta	I-protein
with	O
a	O
lipid	O
raft-associated	O
inhibitor	O
of	O
kappa	B-protein
B	I-protein
factor	I-protein
kinase	I-protein
(	I-protein
IKK	I-protein
)	I-protein
complex	I-protein
plays	O
a	O
role	O
in	O
the	O
activation	O
of	O
the	O
NF-kappa	O
B	O
cascade	O
by	O
TCR	B-protein
and	O
CD28	B-protein
.	O

We	O
investigated	O
the	O
role	O
of	O
protein	B-protein
kinase	I-protein
C	I-protein
theta	I-protein
(	O
PKCtheta	B-protein
)	O
in	O
the	O
activation	O
of	O
the	O
NF-kappaB	B-protein
cascade	O
in	O
primary	B-cell_type
human	I-cell_type
CD4	I-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
lymphocytes	I-cell_type
.	O

Among	O
six	O
or	O
so	O
PKC	B-protein
isoforms	I-protein
expressed	O
in	O
T	B-cell_type
cells	I-cell_type
,	O
only	O
PKCtheta	B-protein
participates	O
in	O
the	O
assembly	O
of	O
the	O
supramolecular	O
activation	O
clusters	O
at	O
the	O
contact	O
site	O
of	O
the	O
TCR	B-protein
with	O
Ag	B-protein
.	O

Signaling	O
via	O
both	O
the	O
TCR	B-protein
and	O
CD28	B-protein
is	O
required	O
for	O
optimal	O
activation	O
of	O
the	O
multisubunit	B-protein
IkappaB	I-protein
kinase	I-protein
(	I-protein
IKK	I-protein
)	I-protein
complex	I-protein
in	O
primary	B-cell_type
human	I-cell_type
T	I-cell_type
lymphocytes	I-cell_type
;	O
this	O
activation	O
could	O
be	O
inhibited	O
by	O
a	O
Ca	O
(	O
2+	O
)	O
-independent	O
PKC	O
isoform	O
inhibitor	O
,	O
rottlerin	O
.	O

Moreover	O
,	O
endogenous	O
PKCtheta	B-protein
physically	O
associates	O
with	O
activated	O
IKK	B-protein
complexes	I-protein
in	O
CD3/CD28-costimulated	B-cell_type
primary	I-cell_type
CD4	I-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
T	I-cell_type
cells	I-cell_type
.	O

The	O
same	O
set	O
of	O
stimuli	O
also	O
induced	O
relocation	O
of	O
endogenous	O
PKCtheta	B-protein
and	O
IKKs	B-protein
to	O
a	O
GM1	O
ganglioside-enriched	O
,	O
detergent-insoluble	O
membrane	O
compartment	O
in	O
primary	B-cell_type
T	I-cell_type
cells	I-cell_type
.	O

IKKs	B-protein
recruited	O
to	O
these	O
lipid	O
rafts	O
were	O
capable	O
of	O
phosphorylating	O
a	O
recombinant	O
IkappaBalpha	O
sustrate	O
.	O

Confocal	O
microscopy	O
further	O
demonstrated	O
that	O
exogenously	O
expressed	O
PKCtheta	B-protein
and	O
IKKss	B-protein
colocalize	O
in	O
the	O
membrane	O
of	O
CD3/CD28-costimulated	B-cell_line
Jurkat	I-cell_line
T	I-cell_line
cells	I-cell_line
.	O

Constitutively	O
active	O
but	O
not	O
kinase-inactive	B-protein
PKCtheta	I-protein
activated	O
IKKbeta	B-protein
in	O
Jurkat	B-cell_line
T	I-cell_line
cells	I-cell_line
.	O

Expression	O
of	O
dominant-active	B-protein
PKCtheta	I-protein
also	O
had	O
stimulatory	O
effects	O
on	O
the	O
CD28	B-DNA
response	I-DNA
element	I-DNA
of	O
the	O
IL-2	B-DNA
promoter	I-DNA
.	O

Taken	O
together	O
,	O
these	O
data	O
show	O
that	O
the	O
activation	O
of	O
PKCtheta	B-protein
by	O
the	O
TCR	B-protein
and	O
CD28	B-protein
plays	O
an	O
important	O
role	O
in	O
the	O
assembly	O
and	O
activation	O
of	O
IKK	B-protein
complexes	I-protein
in	O
the	O
T	O
cell	O
membrane	O

-DOCSTART-	O

T-cell	B-cell_type
-mediated	O
regulation	O
of	O
osteoclastogenesis	O
by	O
signalling	O
cross-talk	O
between	O
RANKL	B-protein
and	O
IFN-gamma	B-protein
.	O

Bone	O
resorption	O
is	O
regulated	O
by	O
the	O
immune	O
system	O
,	O
where	O
T-cell	B-cell_type
expression	O
of	O
RANKL	B-protein
(	O
receptor	B-protein
activator	I-protein
of	I-protein
nuclear	I-protein
factor	I-protein
(	I-protein
NF	I-protein
)	I-protein
-kappaB	I-protein
ligand	I-protein
)	O
,	O
a	O
member	O
of	O
the	O
tumour-necrosis	B-protein
factor	I-protein
family	I-protein
that	O
is	O
essential	O
for	O
osteoclastogenesis	O
,	O
may	O
contribute	O
to	O
pathological	O
conditions	O
,	O
such	O
as	O
autoimmune	O
arthritis	O
.	O

However	O
,	O
whether	O
activated	B-cell_type
T	I-cell_type
cells	I-cell_type
maintain	O
bone	O
homeostasis	O
by	O
counterbalancing	O
the	O
action	O
of	O
RANKL	B-protein
remains	O
unknown	O
.	O

Here	O
we	O
show	O
that	O
T-cell	B-cell_type
production	O
of	O
interferon	B-protein
(	I-protein
IFN	I-protein
)	I-protein
-gamma	I-protein
strongly	O
suppresses	O
osteoclastogenesis	O
by	O
interfering	O
with	O
the	O
RANKL	B-protein
-RANK	O
signalling	O
pathway	O
.	O

IFN-gamma	B-protein
induces	O
rapid	O
degradation	O
of	O
the	O
RANK	B-protein
adapter	I-protein
protein	I-protein
,	O
TRAF6	B-protein
(	O
tumour	B-protein
necrosis	I-protein
factor	I-protein
receptor-associated	I-protein
factor	I-protein
6	I-protein
)	O
,	O
which	O
results	O
in	O
strong	O
inhibition	O
of	O
the	O
RANKL	B-protein
-induced	O
activation	O
of	O
the	O
transcription	B-protein
factor	I-protein
NF-kappaB	B-protein
and	O
JNK	B-protein
.	O

This	O
inhibition	O
of	O
osteoclastogenesis	O
is	O
rescued	O
by	O
overexpressing	O
TRAF6	B-protein
in	O
precursor	B-cell_type
cells	I-cell_type
,	O
which	O
indicates	O
that	O
TRAF6	B-protein
is	O
the	O
target	O
critical	O
for	O
the	O
IFN-gamma	B-protein
action	O
.	O

Furthermore	O
,	O
we	O
provide	O
evidence	O
that	O
the	O
accelerated	O
degradation	O
of	O
TRAF6	B-protein
requires	O
both	O
its	O
ubiquitination	O
,	O
which	O
is	O
initiated	O
by	O
RANKL	B-protein
,	O
and	O
IFN-gamma	B-protein
-induced	O
activation	O
of	O
the	O
ubiquitin-proteasome	O
system	O
.	O

Our	O
study	O
shows	O
that	O
there	O
is	O
cross-talk	O
between	O
the	O
tumour	B-protein
necrosis	I-protein
factor	I-protein
and	O
IFN	B-protein
families	I-protein
of	O
cytokines	B-protein
,	O
through	O
which	O
IFN-gamma	B-protein
provides	O
a	O
negative	O
link	O
between	O
T-cell	B-cell_type
activation	O
and	O
bone	O
resorption	O
.	O

Our	O
results	O
may	O
offer	O
a	O
therapeutic	O
approach	O
to	O
treat	O
the	O
inflammation-induced	O
tissue	O
breakdown	O
.	O

-DOCSTART-	O

Stromal-derived	B-protein
factor	I-protein
1	I-protein
and	O
thrombopoietin	B-protein
regulate	O
distinct	O
aspects	O
of	O
human	O
megakaryopoiesis	O
.	O

The	O
role	O
of	O
the	O
chemokine	B-protein
binding	O
stromal-derived	B-protein
factor	I-protein
1	I-protein
(	O
SDF-1	B-protein
)	O
in	O
normal	O
human	O
megakaryopoiesis	O
at	O
the	O
cellular	O
and	O
molecular	O
levels	O
and	O
its	O
comparison	O
with	O
that	O
of	O
thrombopoietin	B-protein
(	O
TPO	B-protein
)	O
have	O
not	O
been	O
determined	O
.	O

In	O
this	O
study	O
it	O
was	O
found	O
that	O
SDF-1	B-protein
,	O
unlike	O
TPO	B-protein
,	O
does	O
not	O
stimulate	O
alpha	O
(	O
IIb	O
)	O
beta	O
(	O
3	O
)	O
(	O
+	O
)	O
cell	O
proliferation	O
or	O
differentiation	O
or	O
have	O
an	O
antiapoptotic	O
effect	O
.	O

However	O
,	O
it	O
does	O
induce	O
chemotaxis	O
,	O
trans-Matrigel	O
migration	O
,	O
and	O
secretion	O
of	O
matrix	O
metalloproteinase	B-protein
9	I-protein
(	O
MMP-9	B-protein
)	O
and	O
vascular	B-protein
endothelial	I-protein
growth	I-protein
factor	I-protein
(	O
VEGF	B-protein
)	O
by	O
these	O
cells	O
,	O
and	O
both	O
SDF-1	B-protein
and	O
TPO	B-protein
increase	O
the	O
adhesion	O
of	O
alpha	B-cell_type
(	I-cell_type
IIb	I-cell_type
)	I-cell_type
beta	I-cell_type
(	I-cell_type
3	I-cell_type
)	I-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
cells	I-cell_type
to	O
fibrinogen	B-protein
and	O
vitronectin	B-protein
.	O

Investigating	O
the	O
intracellular	O
signaling	O
pathways	O
induced	O
by	O
SDF-1	B-protein
and	O
TPO	B-protein
revealed	O
some	O
overlapping	O
patterns	O
of	O
protein	O
phosphorylation/activation	O
(	O
mitogen-activated	B-protein
protein	I-protein
kinase	I-protein
[	I-protein
MAPK	I-protein
]	I-protein
p42/44	I-protein
,	O
MAPK	B-protein
p38	I-protein
,	O
and	O
AKT	B-protein
[	I-protein
protein	I-protein
kinase	I-protein
B	I-protein
]	I-protein
)	O
and	O
some	O
that	O
were	O
distinct	O
for	O
TPO	B-protein
(	O
eg	O
,	O
JAK-STAT	B-protein
)	O
and	O
for	O
SDF-1	B-protein
(	O
eg	O
,	O
NF-kappa	B-protein
B	I-protein
)	O
.	O

It	O
was	O
also	O
found	O
that	O
though	O
inhibition	O
of	O
phosphatidyl-inositol	B-protein
3-kinase	I-protein
(	O
PI-3K	B-protein
)	O
by	O
LY294002	O
in	O
alpha	B-cell_type
(	I-cell_type
IIb	I-cell_type
)	I-cell_type
beta	I-cell_type
(	I-cell_type
3	I-cell_type
)	I-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
cells	I-cell_type
induced	O
apoptosis	O
and	O
inhibited	O
chemotaxis	O
adhesion	O
and	O
the	O
secretion	O
of	O
MMP-9	B-protein
and	O
VEGF	B-protein
,	O
the	O
inhibition	O
of	O
MAPK	B-protein
p42/44	I-protein
(	O
by	O
the	O
MEK	O
inhibitor	O
U0126	O
)	O
had	O
no	O
effect	O
on	O
the	O
survival	O
,	O
proliferation	O
,	O
and	O
migration	O
of	O
these	O
cells	O
.	O

Hence	O
,	O
it	O
is	O
suggested	O
that	O
the	O
proliferative	O
effect	O
of	O
TPO	B-protein
is	O
more	O
related	O
to	O
activation	O
of	O
the	O
JAK-STAT	O
pathway	O
(	O
unique	O
to	O
TPO	B-protein
)	O
,	O
and	O
the	O
PI-3K-AKT	O
axis	O
is	O
differentially	O
involved	O
in	O
TPO-	O
and	O
SDF-1-dependent	O
signaling	O
.	O

Accordingly	O
,	O
PI-3K	B-protein
is	O
involved	O
in	O
TPO	B-protein
-mediated	O
inhibition	O
of	O
apoptosis	O
,	O
TPO-	O
and	O
SDF-1-regulated	O
adhesion	O
to	O
fibrinogen	B-protein
and	O
vitronectin	B-protein
,	O
and	O
SDF-1	B-protein
-mediated	O
migration	O
.	O

This	O
study	O
expands	O
the	O
understanding	O
of	O
the	O
role	O
of	O
SDF-1	B-protein
and	O
TPO	B-protein
in	O
normal	O
human	O
megakaryopoiesis	O
and	O
indicates	O
the	O
molecular	O
basis	O
of	O
the	O
observed	O
differences	O
in	O
cellular	O
responses	O
.	O

(	O
Blood.	O
2000	O
;	O
96	O
:	O
4142-4151	O
)	O

-DOCSTART-	O

Adhesion	O
of	O
immature	B-cell_type
and	I-cell_type
mature	I-cell_type
T	I-cell_type
cells	I-cell_type
induces	O
in	O
human	B-cell_type
thymic	I-cell_type
epithelial	I-cell_type
cells	I-cell_type
(	O
TEC	B-cell_type
)	O
activation	O
of	O
IL-6	B-protein
gene	I-protein
trascription	I-protein
factors	I-protein
(	O
NF-kappaB	B-protein
and	O
NF-IL6	B-protein
)	O
and	O
IL-6	O
gene	O
expression	O
:	O
role	O
of	O
alpha3beta1	B-protein
and	I-protein
alpha6beta4	I-protein
integrins	I-protein
.	O

T	B-cell_type
cell	I-cell_type
precursors	I-cell_type
homed	O
to	O
thymus	O
develop	O
in	O
close	O
contact	O
with	O
stromal	B-cell_type
cells	I-cell_type
.	O

Among	O
them	O
,	O
thymic	B-cell_type
epithelial	I-cell_type
cells	I-cell_type
(	O
TEC	B-cell_type
)	O
are	O
known	O
to	O
exert	O
dominant	O
roles	O
in	O
their	O
survival	O
and	O
functional	O
shaping	O
.	O

Key	O
molecules	O
mediating	O
TEC	B-cell_type
/	O
thymocytes	B-cell_type
interactions	O
include	O
cytokines	B-protein
and	O
growth	B-protein
factors	I-protein
secreted	O
by	O
the	O
two	O
cell	O
types	O
and	O
adhesion	B-protein
receptors	I-protein
mediating	O
cell	O
contact	O
.	O

Signaling	O
events	O
triggered	O
in	O
thymocytes	B-cell_type
by	O
adhesion	O
to	O
epithelial	B-cell_type
cells	I-cell_type
have	O
been	O
extensively	O
investigated	O
,	O
whereas	O
little	O
is	O
known	O
on	O
the	O
opposite	O
phenomenon	O
.	O

We	O
have	O
previously	O
investigated	O
this	O
issue	O
in	O
a	O
co-culture	O
system	O
composed	O
of	O
TEC	B-cell_type
cultures	I-cell_type
derived	O
from	O
human	O
normal	O
thymus	O
and	O
heterologous	B-cell_type
thymocytes	I-cell_type
.	O

We	O
demonstrated	O
that	O
thymocytes	B-cell_type
adhere	O
to	O
TEC	B-cell_type
involving	O
beta1	B-protein
and	I-protein
beta4	I-protein
integrins	I-protein
and	O
induce	O
the	O
clustering	O
of	O
alpha3beta1	B-protein
and	I-protein
alpha6beta4	I-protein
heterodimers	I-protein
at	O
the	O
TEC	O
surface	O
.	O

In	O
addition	O
thymocyte	O
adhesion	O
was	O
followed	O
by	O
activation	O
of	O
NF-kappaB	B-protein
and	O
NF-IL6	B-protein
gene	B-protein
transcription	I-protein
factors	I-protein
and	O
enhanced	O
IL-6	O
production	O
.	O

The	O
two	O
latter	O
phenomena	O
were	O
reproduced	O
by	O
the	O
cross-linking	O
of	O
the	O
alpha3	B-protein
,	O
alpha6	B-protein
,	O
beta1	B-protein
and	I-protein
beta4	I-protein
integrins	I-protein
,	O
thus	O
implying	O
that	O
the	O
alpha3beta1	B-protein
and	I-protein
alpha6beta4	I-protein
heterodimers	I-protein
can	O
signal	O
during	O
thymocyte	O
adhesion	O
.	O

We	O
have	O
extended	O
our	O
previous	O
work	O
investigating	O
in	O
the	O
same	O
experimental	O
setting	O
the	O
inducing	O
activity	O
of	O
non	O
stimulated	O
or	O
activated	O
policlonal	B-cell_type
or	I-cell_type
clonal	I-cell_type
mature	I-cell_type
T	I-cell_type
cells	I-cell_type
as	O
representative	O
of	O
the	O
more	O
mature	B-cell_type
thymocyte	I-cell_type
subset	I-cell_type
.	O

We	O
found	O
that	O
adhesion	O
of	O
unstimulated	B-cell_type
T	I-cell_type
cell	I-cell_type
i	O
)	O
involved	O
beta1	O
,	O
but	O
not	O
beta4	O
integrin	O
functions	O
at	O
the	O
surface	O
ii	O
)	O
induced	O
the	O
clustering	O
of	O
alpha3beta1	B-protein
,	O
but	O
not	O
alpha2beta1	B-protein
heterodimers	I-protein
at	O
the	O
TEC	O
surface	O
and	O
iii	O
)	O
up-regulated	O
the	O
nuclear	O
binding	O
activity	O
of	O
NF-kappaB	B-protein
transcription	I-protein
factor	I-protein
and	O
the	O
IL-6	O
secretion	O
.	O

We	O
propose	O
that	O
alpha3beta1	B-protein
and	I-protein
alpha6beta4	I-protein
heterodimers	I-protein
are	O
induced	O
to	O
cluster	O
at	O
the	O
TEC	O
surface	O
recognizing	O
yet	O
unknown	O
cellular	B-protein
ligands	I-protein
differentially	O
expressed	O
during	O
T	O
cell	O
development	O
.	O

-DOCSTART-	O

Identification	O
and	O
characterization	O
of	O
SKAT-2	B-DNA
,	O
a	O
novel	O
Th2-specific	B-DNA
zinc	I-DNA
finger	I-DNA
gene	I-DNA
.	O

We	O
have	O
identified	O
a	O
novel	O
Kruppel-type	B-DNA
zinc	I-DNA
finger	I-DNA
(	I-DNA
ZF	I-DNA
)	I-DNA
gene	I-DNA
,	O
SKAT-2	B-DNA
,	O
which	O
is	O
selectively	O
expressed	O
by	O
murine	B-cell_type
Th2	I-cell_type
cells	I-cell_type
.	O

The	O
protein	O
encoded	O
by	O
this	O
gene	O
has	O
14	O
C2H2-type	O
ZF	O
tandemly	O
arrayed	O
at	O
its	O
C	B-DNA
terminus	I-DNA
and	O
N-terminal	B-DNA
SCAN	B-DNA
box	I-DNA
and	O
KRAB	B-DNA
domains	I-DNA
.	O

SKAT-2	B-DNA
is	O
tissue	O
restricted	O
in	O
expression	O
at	O
the	O
RNA	O
level	O
,	O
detectable	O
only	O
in	O
brain	O
and	O
at	O
low	O
levels	O
in	O
kidney	O
and	O
spleen	O
and	O
few	O
hematopoietic	B-cell_line
cell	I-cell_line
lines	I-cell_line
.	O

By	O
in	O
situ	O
hybridization	O
,	O
SKAT-2	B-DNA
expression	O
was	O
found	O
to	O
peak	O
in	O
antigen-stimulated	O
CD4	B-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
T	I-cell_type
cells	I-cell_type
after	O
2-3	O
days	O
of	O
culture	O
under	O
Th2	O
but	O
not	O
Th1	O
biasing	O
conditions	O
.	O

This	O
pattern	O
of	O
expression	O
closely	O
mirrored	O
that	O
of	O
GATA-3	B-protein
in	O
the	O
same	O
cells	O
.	O

In	O
transient	O
transfection	O
experiments	O
in	O
phorbol	O
12-myristate	O
13-acetate/ionomycin-stimulated	O
EL4	B-cell_line
cells	I-cell_line
,	O
SKAT-2	B-DNA
was	O
found	O
to	O
up-regulate	O
the	O
activity	O
of	O
the	O
IL-4	B-DNA
but	I-DNA
not	I-DNA
the	I-DNA
IL-5	I-DNA
promoter	I-DNA
,	O
contrasting	O
with	O
the	O
ability	O
of	O
GATA-3	B-protein
to	O
activate	O
both	O
promoters	B-DNA
.	O

This	O
result	O
was	O
confirmed	O
using	O
clones	O
of	O
EL4	B-cell_line
cells	I-cell_line
stably	O
expressing	O
an	O
inducible	O
form	O
of	O
SKAT-2	B-DNA
,	O
thus	O
SKAT-2	B-DNA
is	O
a	O
novel	O
Th2-specific	B-DNA
gene	I-DNA
that	O
may	O
play	O
a	O
role	O
in	O
selective	O
regulation	O
of	O
cytokine	B-DNA
genes	I-DNA
in	O
T	B-cell_type
cells	I-cell_type
.	O

-DOCSTART-	O

hsp70	B-protein
interacting	I-protein
protein	I-protein
Hip	I-protein
does	O
not	O
affect	O
glucocorticoid	B-protein
receptor	I-protein
folding	O
by	O
the	O
hsp90-based	O
chaperone	O
machinery	O
except	O
to	O
oppose	O
the	O
effect	O
of	O
BAG-1	B-protein
.	O

Reticulocyte	O
lysate	O
contains	O
a	O
chaperone	O
system	O
that	O
assembles	O
glucocorticoid	B-protein
receptor	I-protein
(	I-protein
GR	I-protein
)	I-protein
.hsp90	I-protein
heterocomplexes	I-protein
.	O

Using	O
purified	B-protein
proteins	I-protein
,	O
we	O
have	O
prepared	O
a	O
five-protein	O
heterocomplex	O
assembly	O
system	O
consisting	O
of	O
two	O
proteins	O
essential	O
for	O
heterocomplex	O
assembly-	O
hsp90	B-protein
and	O
hsp70	B-protein
-and	O
three	O
proteins	O
that	O
act	O
as	O
co-chaperones	O
to	O
enhance	O
assembly-	O
Hop	B-protein
,	O
hsp40	B-protein
,	O
p23	B-protein
[	O
Morishima	O
,	O
Y.	O
,	O
Kanelakis	O
,	O
K.	O
C.	O
,	O
Silverstein	O
,	O
A.	O
M.	O
,	O
Dittmar	O
,	O
K.	O
D.	O
,	O
Estrada	O
,	O
L.	O
,	O
and	O
Pratt	O
,	O
W.	O
B.	O
(	O
2000	O
)	O
J.	O
Biol.	O
Chem.	O
275	O
,	O
6894-6900	O
]	O
.	O

The	O
hsp70	B-protein
co-chaperone	I-protein
Hip	I-protein
has	O
been	O
recovered	O
in	O
receptor.hsp90	B-protein
heterocomplexes	I-protein
at	O
an	O
intermediate	O
stage	O
of	O
assembly	O
in	O
reticulocyte	O
lysate	O
,	O
and	O
Hip	B-protein
is	O
also	O
thought	O
to	O
be	O
an	O
intrinsic	O
component	O
of	O
the	O
assembly	O
machinery	O
.	O

Here	O
we	O
show	O
that	O
immunodepletion	O
of	O
Hip	B-protein
from	O
reticulocyte	O
lysate	O
or	O
addition	O
of	O
high	O
levels	O
of	O
Hip	B-protein
to	O
the	O
purified	O
five-protein	O
system	O
does	O
not	O
affect	O
GR.hsp90	O
heterocomplex	O
assembly	O
or	O
the	O
activation	O
of	O
steroid	O
binding	O
activity	O
that	O
occurs	O
with	O
assembly	O
.	O

Despite	O
the	O
fact	O
that	O
Hip	B-protein
does	O
not	O
affect	O
assembly	O
,	O
it	O
is	O
recovered	O
in	O
GR.hsp90	B-protein
heterocomplexes	I-protein
assembled	O
by	O
both	O
systems	O
.	O

In	O
the	O
five-protein	O
system	O
,	O
Hip	B-protein
prevents	O
inhibition	O
of	O
assembly	O
by	O
the	O
hsp70	B-protein
co-chaperone	I-protein
BAG-1	I-protein
,	O
and	O
cotransfection	O
of	O
Hip	B-protein
with	O
BAG-1	B-protein
opposes	O
BAG-1	B-protein
reduction	O
of	O
steroid	O
binding	O
activity	O
in	O
COS	B-cell_line
cells	I-cell_line
.	O

We	O
conclude	O
that	O
Hip	B-protein
is	O
not	O
a	O
component	O
of	O
the	O
assembly	O
machinery	O
but	O
that	O
it	O
could	O
play	O
a	O
regulatory	O
role	O
in	O
opposition	O
to	O
BAG-1	B-protein
.	O

-DOCSTART-	O

Cutting	O
edge	O
:	O
STAT6-deficient	O
mice	O
have	O
enhanced	O
tumor	O
immunity	O
to	O
primary	O
and	O
metastatic	O
mammary	O
carcinoma	O
.	O

STAT4	B-protein
and	O
STAT6	B-protein
are	O
essential	O
for	O
the	O
development	O
of	O
CD4	O
(	O
+	O
)	O
Th1	O
and	O
Th2	O
development	O
,	O
respectively	O
.	O

Tumor	O
immunologists	O
have	O
hypothesized	O
that	O
Th1	B-cell_type
cells	I-cell_type
are	O
critical	O
in	O
tumor	O
immunity	O
because	O
they	O
facilitate	O
differentiation	O
of	O
CD8	B-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
T	I-cell_type
cells	I-cell_type
,	O
which	O
are	O
potent	O
anti-tumor	B-cell_type
effectors	I-cell_type
.	O

We	O
have	O
used	O
STAT4	O
(	O
-/-	O
)	O
and	O
STAT6	O
(	O
-/-	O
)	O
mice	O
to	O
test	O
this	O
hypothesis	O
.	O

BALB/c	O
and	O
knockout	O
mice	O
were	O
challenged	O
in	O
the	O
mammary	O
gland	O
with	O
the	O
highly	O
malignant	O
and	O
spontaneously	O
metastatic	O
BALB/c-derived	O
4T1	O
mammary	O
carcinoma	O
.	O

Primary	O
tumor	O
growth	O
and	O
metastatic	O
disease	O
are	O
reduced	O
in	O
STAT6	O
(	O
-/-	O
)	O
mice	O
relative	O
to	O
BALB/c	O
and	O
STAT4	O
(	O
-/-	O
)	O
mice	O
.	O

Ab	O
depletions	O
demonstrate	O
that	O
the	O
effect	O
is	O
mediated	O
by	O
CD8	B-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
T	I-cell_type
cells	I-cell_type
,	O
and	O
immunized	O
STAT6	O
(	O
-/-	O
)	O
mice	O
have	O
higher	O
levels	O
of	O
4T1-specific	B-cell_type
CTL	I-cell_type
than	O
BALB/c	O
or	O
STAT4	O
(	O
-/-	O
)	O
mice	O
.	O

Surprisingly	O
,	O
Th1	B-cell_type
or	I-cell_type
Th2	I-cell_type
cells	I-cell_type
are	O
not	O
involved	O
,	O
because	O
CD4	O
depletion	O
does	O
not	O
diminish	O
the	O
anti-tumor	O
effect	O
.	O

Therefore	O
,	O
deletion	O
of	O
the	O
STAT6	B-DNA
gene	I-DNA
facilitates	O
development	O
of	O
potent	O
anti-tumor	O
immunity	O
via	O
a	O
CD4	O
(	O
+	O
)	O
-independent	O
pathway	O
.	O

-DOCSTART-	O

Activation	O
of	O
oncogenic	B-protein
transcription	I-protein
factor	I-protein
AP-1	I-protein
in	O
T	B-cell_type
cells	I-cell_type
infected	O
with	O
human	O
T	O
cell	O
leukemia	O
virus	O
type	O
1	O
.	O

Human	O
T	O
cell	O
leukemia	O
virus	O
type	O
1	O
(	O
HTLV-1	O
)	O
Tax	B-protein
protein	I-protein
transforms	O
primary	B-cell_type
human	I-cell_type
T	I-cell_type
cells	I-cell_type
in	O
vitro	O
.	O

We	O
previously	O
showed	O
that	O
Tax	B-protein
induces	O
the	O
expression	O
of	O
various	O
family	O
members	O
of	O
the	O
transcription	B-protein
factor	I-protein
AP-1	I-protein
such	O
as	O
c-Jun	B-protein
,	O
JunD	B-protein
,	O
c-Fos	B-protein
,	O
and	O
Fra-1	B-protein
at	O
the	O
mRNA	O
level	O
in	O
T	B-cell_type
cells	I-cell_type
.	O

In	O
this	O
study	O
,	O
we	O
have	O
examined	O
the	O
ability	O
of	O
Tax	B-protein
to	O
activate	O
transcription	O
through	O
the	O
AP-1-binding	B-DNA
site	I-DNA
(	O
AP-1	B-DNA
site	I-DNA
)	O
.	O

A	O
transient	O
transfection	O
study	O
showed	O
that	O
Tax	B-protein
can	O
activate	O
transcription	O
through	O
the	O
AP-1-binding	B-DNA
site	I-DNA
in	O
a	O
human	B-cell_line
T	I-cell_line
cell	I-cell_line
line	I-cell_line
,	O
whereas	O
any	O
combination	O
of	O
AP-1	B-protein
proteins	I-protein
did	O
so	O
much	O
less	O
than	O
Tax	B-protein
,	O
indicating	O
that	O
the	O
activation	O
of	O
the	O
AP-1	B-DNA
site	I-DNA
by	O
Tax	B-protein
may	O
require	O
a	O
mechanism	O
other	O
than	O
the	O
induction	O
of	O
AP-1	B-RNA
mRNA	I-RNA
.	O

Fresh	B-cell_type
peripheral	I-cell_type
blood	I-cell_type
leukemia	I-cell_type
cells	I-cell_type
of	O
all	O
surveyed	O
ATL	O
patients	O
displayed	O
constitutive	O
AP-1	O
DNA-binding	O
activity	O
,	O
whereas	O
no	O
normal	O
individuals	O
did	O
.	O

However	O
,	O
the	O
HTLV-1	B-DNA
genes	I-DNA
,	O
including	O
tax	B-DNA
,	O
are	O
not	O
significantly	O
expressed	O
in	O
fresh	O
leukemia	B-cell_type
cells	I-cell_type
from	O
ATL	O
patients	O
.	O

Our	O
present	O
results	O
suggest	O
that	O
activation	O
of	O
AP-1	B-protein
occurs	O
through	O
Tax-dependent	O
and	O
-independent	O
mechanisms	O
in	O
HTLV-1-infected	O
T	B-cell_type
cells	I-cell_type
,	O
which	O
may	O
play	O
some	O
roles	O
in	O
dysregulated	O
phenotypes	O
of	O
HTLV-1-infected	B-cell_type
cells	I-cell_type
.	O

-DOCSTART-	O

2	O
,	O
2	O
'	O
,	O
4	O
,	O
6	O
,	O
6'-pentachlorobiphenyl	O
induces	O
apoptosis	O
in	O
human	B-cell_type
monocytic	I-cell_type
cells	I-cell_type
.	O

Polychlorinatedbiphenyls	O
(	O
PCBs	O
)	O
are	O
a	O
group	O
of	O
persistent	O
and	O
widely	O
dispersed	O
environmental	O
pollutants	O
,	O
some	O
of	O
which	O
may	O
be	O
immunotoxic	O
.	O

In	O
the	O
present	O
study	O
,	O
we	O
investigated	O
the	O
effect	O
of	O
PCBs	O
on	O
immune	O
system	O
by	O
assessing	O
apoptotic	O
cell	O
death	O
in	O
human	B-cell_line
monocytic	I-cell_line
U937	I-cell_line
cells	I-cell_line
.	O

Among	O
the	O
various	O
congeners	O
tested	O
,	O
2	O
,	O
2	O
'	O
,	O
4	O
,	O
6	O
,	O
6'-pentachlorobiphenyl	O
(	O
PeCB	O
)	O
,	O
a	O
highly	O
ortho-substituted	O
congener	O
,	O
specifically	O
induced	O
DNA	O
fragmentation	O
,	O
a	O
hallmark	O
of	O
apoptosis	O
,	O
while	O
the	O
other	O
examined	O
di-	O
,	O
tri-	O
,	O
tetra-	O
,	O
and	O
pentachlorobiphenyls	O
did	O
not	O
.	O

To	O
further	O
study	O
the	O
2	O
,	O
2	O
'	O
,	O
4	O
,	O
6	O
,	O
6'-PeCB-induced	O
cell	O
death	O
,	O
various	O
features	O
of	O
apoptosis	O
were	O
examined	O
.	O

2	O
,	O
2	O
'	O
,	O
4	O
,	O
6	O
,	O
6'-PeCB	O
caused	O
a	O
decrease	O
in	O
cell	O
viability	O
and	O
induced	O
cellular	O
morphologic	O
features	O
characteristic	O
of	O
apoptosis	O
such	O
as	O
chromatin	O
aggregation	O
and	O
apoptotic	O
bodies	O
.	O

In	O
addition	O
,	O
caspase-3	B-protein
,	O
an	O
executioner	O
of	O
apoptosis	O
,	O
was	O
activated	O
and	O
its	O
substrate	O
,	O
poly	B-protein
(	I-protein
ADP-ribose	I-protein
)	I-protein
polymerase	I-protein
(	O
PARP	B-protein
)	O
,	O
was	O
cleaved	O
during	O
2	O
,	O
2	O
'	O
,	O
4	O
,	O
6	O
,	O
6'-PeCB-induced	O
apoptosis	O
.	O

In	O
contrast	O
,	O
3	O
,	O
3	O
'	O
,	O
4	O
,	O
4	O
'	O
,	O
5-PeCB	O
,	O
a	O
congener	O
of	O
coplanar	O
structure	O
,	O
as	O
well	O
as	O
2	O
,	O
3	O
,	O
7	O
,	O
8-TCDD	O
did	O
not	O
induce	O
apoptosis	O
in	O
these	O
human	B-cell_type
monocytic	I-cell_type
cells	I-cell_type
,	O
although	O
they	O
potently	O
induced	O
CYP	B-protein
1A1	I-protein
in	O
human	B-cell_line
hepatoma	I-cell_line
Hep	I-cell_line
G2	I-cell_line
cells	I-cell_line
.	O

Taken	O
together	O
,	O
the	O
data	O
indicate	O
that	O
2	O
,	O
2	O
'	O
,	O
4	O
,	O
6	O
,	O
6'-PeCB	O
induces	O
apoptosis	O
in	O
human	B-cell_type
monocytic	I-cell_type
cells	I-cell_type
through	O
a	O
mechanism	O
that	O
is	O
independent	O
of	O
the	O
arylhydrocarbon	B-protein
receptor	I-protein
.	O

This	O
suggests	O
a	O
possibly	O
separate	O
mechanism	O
by	O
which	O
PCBs	O
cause	O
immunosuppression	O
.	O

-DOCSTART-	O

Effects	O
of	O
deregulated	O
Raf	O
activation	O
on	O
integrin	B-protein
,	O
cytokine-receptor	O
expression	O
and	O
the	O
induction	O
of	O
apoptosis	O
in	O
hematopoietic	B-cell_type
cells	I-cell_type
.	O

The	O
effects	O
of	O
deregulated	O
Raf	O
activation	O
on	O
the	O
growth	O
and	O
differentiation	O
of	O
hematopoietic	B-cell_type
cells	I-cell_type
were	O
investigated	O
.	O

The	O
cytokine-dependent	B-cell_line
murine	I-cell_line
myeloid	I-cell_line
FDC-P1	I-cell_line
and	O
human	B-cell_line
erythroleukemic	I-cell_line
TF-1	I-cell_line
cell	I-cell_line
lines	I-cell_line
were	O
transformed	O
to	O
grow	O
in	O
response	O
to	O
deregulated	O
Raf	O
expression	O
in	O
the	O
absence	O
of	O
exogenous	B-protein
cytokines	I-protein
.	O

The	O
conditionally	O
active	O
Raf	B-protein
proteins	I-protein
were	O
regulated	O
by	O
beta-estradiol	O
as	O
cDNAs	B-DNA
containing	O
the	O
Raf	B-protein
catalytic	I-protein
,	O
but	O
lacking	O
negative-regulatory	B-protein
domains	I-protein
,	O
were	O
ligated	O
to	O
the	O
hormone	B-protein
binding	I-protein
domain	I-protein
of	O
the	O
estrogen	B-protein
receptor	I-protein
(	O
deltaRaf	B-protein
:	I-protein
ER	I-protein
)	O
.	O

Continuous	O
deltaRaf	B-protein
expression	O
prevented	O
apoptosis	O
in	O
the	O
absence	O
of	O
exogenous	B-protein
cytokines	I-protein
and	O
altered	O
the	O
morphology	O
of	O
the	O
FD/deltaRaf	B-cell_type
:	I-cell_type
ER	I-cell_type
cells	I-cell_type
as	O
they	O
grew	O
in	O
large	O
aggregated	O
masses	O
(	O
>	O
100	O
cells	O
)	O
whereas	O
the	O
parental	B-cell_line
cytokine-dependent	I-cell_line
FDC-P1	I-cell_line
cells	I-cell_line
grew	O
in	O
smaller	O
grape-like	O
clusters	O
(	O
<	O
10	O
cells	O
)	O
.	O

FD/deltaRaf-1	B-cell_line
:	I-cell_line
ER	I-cell_line
cells	I-cell_line
growing	O
in	O
response	O
to	O
Raf	O
activation	O
displayed	O
decreased	O
levels	O
of	O
the	O
Mac-2	B-protein
and	I-protein
Mac-3	I-protein
molecules	I-protein
on	O
their	O
cell	B-cell_type
surface	I-cell_type
.	O

In	O
contrast	O
,	O
when	O
these	O
cells	O
were	O
cultured	O
in	O
IL-3	B-protein
,	O
higher	O
levels	O
of	O
these	O
adhesion	B-protein
molecules	I-protein
were	O
detected	O
.	O

Expression	O
of	O
activated	O
Raf	B-protein
oncoproteins	I-protein
also	O
abrogated	O
cytokine	O
dependency	O
and	O
prevented	O
apoptosis	O
of	O
TF-1	B-cell_line
cells	I-cell_line
.	O

Moreover	O
,	O
the	O
differentiation	O
status	O
of	O
these	O
Raf-responsive	B-cell_type
cells	I-cell_type
was	O
more	O
immature	O
upon	O
Raf	O
activation	O
as	O
culture	O
with	O
the	O
differentiation-inducing	O
agent	O
phorbol	O
12	O
myristate	O
13-acetate	O
(	O
PMA	O
)	O
and	O
beta-estradiol	O
resulted	O
in	O
decreased	O
levels	O
of	O
the	O
CD11b	B-protein
and	I-protein
CD18	I-protein
integrin	I-protein
molecules	I-protein
on	O
the	O
cell	B-cell_type
surface	I-cell_type
.	O

In	O
contrast	O
when	O
the	O
Raf-responsive	B-cell_type
cells	I-cell_type
were	O
induced	O
to	O
differentiate	O
with	O
PMA	O
and	O
GM-CSF	B-protein
,	O
in	O
the	O
absence	O
of	O
deltaRaf	B-protein
:	I-protein
ER	I-protein
activation	O
,	O
increased	O
levels	O
of	O
the	O
CD11b	B-protein
and	I-protein
CD18	I-protein
molecules	I-protein
were	O
detected	O
.	O

Retinoic	O
acid	O
(	O
RA	O
)	O
inhibited	O
3H-thymidine	O
incorporation	O
in	O
response	O
to	O
GM-CSF	B-protein
.	O

Interestingly	O
,	O
Raf	O
activation	O
counterbalanced	O
the	O
inhibition	O
of	O
DNA	O
synthesis	O
caused	O
by	O
RA	O
but	O
not	O
PMA	O
.	O

Thus	O
deregulated	O
Raf	O
expression	O
can	O
alter	O
cytokine	O
dependency	O
,	O
integrin	O
expression	O
and	O
the	O
stage	O
of	O
differentiation	O
.	O

These	O
Raf-responsive	B-cell_line
cell	I-cell_line
lines	I-cell_line
will	O
be	O
useful	O
in	O
elucidating	O
the	O
roles	O
of	O
the	O
MAP	O
kinase	O
cascade	O
on	O
hematopoietic	B-cell_type
cell	I-cell_type
differentiation	O
and	O
malignant	O
transformation	O

-DOCSTART-	O

Cyclic	O
AMP	O
activates	O
p38	B-protein
mitogen-activated	I-protein
protein	I-protein
kinase	I-protein
in	O
Th2	B-cell_type
cells	I-cell_type
:	O
phosphorylation	O
of	O
GATA-3	B-protein
and	O
stimulation	O
of	O
Th2	B-cell_type
cytokine	O
gene	O
expression	O
.	O

cAMP	O
is	O
an	O
important	O
second	O
messenger	O
with	O
immunomodulatory	O
properties	O
.	O

Elevation	O
of	O
intracellular	O
cAMP	O
in	O
T	B-cell_type
cells	I-cell_type
,	O
induced	O
by	O
agents	O
such	O
as	O
IL-1alpha	B-protein
or	O
PGs	B-protein
,	O
inhibits	O
T	O
cell	O
activation	O
.	O

In	O
effector	O
T	B-cell_type
cells	I-cell_type
,	O
an	O
increase	O
in	O
the	O
level	O
of	O
intracellular	O
cAMP	O
inhibits	O
cytokine	B-protein
production	O
in	O
Th1	B-cell_type
cells	I-cell_type
but	O
stimulates	O
cytokine	B-protein
production	O
in	O
Th2	B-cell_type
cells	I-cell_type
.	O

Here	O
we	O
report	O
that	O
cAMP-induced	O
effects	O
in	O
Th2	B-cell_type
cells	I-cell_type
occur	O
independently	O
of	O
the	O
protein	B-protein
kinase	I-protein
A	I-protein
pathway	O
,	O
which	O
is	O
the	O
major	O
mediator	O
of	O
cAMP-induced	O
signaling	O
events	O
in	O
most	O
cell	O
types	O
.	O

Instead	O
,	O
cAMP	O
stimulates	O
activation	O
of	O
p38	B-protein
mitogen-activated	I-protein
protein	I-protein
kinase	I-protein
in	O
Th2	B-cell_type
cells	I-cell_type
.	O

This	O
appears	O
to	O
be	O
a	O
Th2	B-cell_type
-selective	O
event	O
because	O
cAMP	O
barely	O
increased	O
p38	B-protein
phosphorylation	O
in	O
Th1	B-cell_type
cells	I-cell_type
.	O

We	O
show	O
that	O
in	O
Th2	B-cell_type
cells	I-cell_type
,	O
cAMP	O
promotes	O
the	O
production	O
of	O
both	O
IL-5	B-protein
and	O
IL-13	B-protein
,	O
which	O
play	O
distinct	O
but	O
critical	O
roles	O
in	O
asthma	O
pathogenesis	O
.	O

Our	O
data	O
also	O
show	O
that	O
cAMP	O
causes	O
increased	O
phosphorylation	O
of	O
the	O
transcription	B-protein
factor	I-protein
GATA-3	I-protein
,	O
which	O
we	O
have	O
shown	O
is	O
a	O
critical	O
regulator	O
of	O
Th2	B-cell_type
cytokine	O
gene	O
expression	O
and	O
,	O
in	O
turn	O
,	O
of	O
airway	O
inflammation	O
in	O
mice	O
.	O

Thus	O
,	O
Th2	B-cell_type
-specific	O
GATA-3	B-protein
expression	O
and	O
p38	B-protein
mitogen-activated	I-protein
protein	I-protein
kinase	I-protein
activation	O
together	O
provide	O
a	O
molecular	O
basis	O
for	O
the	O
differential	O
effects	O
of	O
cAMP	O
in	O
the	O
two	O
T	B-cell_type
helper	I-cell_type
cell	I-cell_type
subsets	I-cell_type
.	O

-DOCSTART-	O

Characterization	O
of	O
IL-4	B-protein
and	O
IL-13	B-protein
signals	O
dependent	O
on	O
the	O
human	B-protein
IL-13	I-protein
receptor	I-protein
alpha	I-protein
chain	I-protein
1	I-protein
:	O
redundancy	O
of	O
requirement	O
of	O
tyrosine	O
residue	O
for	O
STAT3	O
activation	O
.	O

IL-4	B-protein
and	O
IL-13	B-protein
are	O
pleiotropic	B-protein
cytokines	I-protein
whose	O
biological	O
activities	O
overlap	O
with	O
each	O
other	O
.	O

IL-13	B-protein
receptor	I-protein
alpha	I-protein
chain	I-protein
1	I-protein
(	O
IL-13R	B-protein
alpha	I-protein
1	I-protein
)	O
is	O
necessary	O
for	O
binding	O
to	O
IL-13	B-protein
,	O
and	O
the	O
heterodimer	O
composed	O
of	O
IL-13R	B-protein
alpha	I-protein
1	I-protein
and	O
IL-4R	B-protein
alpha	I-protein
chain	I-protein
transduces	O
IL-13	B-protein
and	O
IL-4	B-protein
signals	O
;	O
however	O
,	O
the	O
functional	O
mapping	O
of	O
the	O
intracellular	B-protein
domain	I-protein
of	O
IL-13R	B-protein
alpha	I-protein
1	I-protein
is	O
not	O
fully	O
understood	O
.	O

In	O
this	O
study	O
,	O
we	O
constructed	O
wild	O
and	O
mutated	O
types	O
of	O
human	B-protein
IL-13R	I-protein
alpha	I-protein
1	I-protein
,	O
and	O
analyzed	O
IL-4	B-protein
and	O
IL-13	B-protein
signals	O
using	O
an	O
IL-13R	B-protein
alpha	I-protein
1	I-protein
-transfected	O
human	B-cell_line
B	I-cell_line
cell	I-cell_line
line	I-cell_line
.	O

Expression	O
of	O
IL-13R	B-protein
alpha	I-protein
1	I-protein
evoked	O
STAT3	B-protein
activation	O
by	O
IL-4	B-protein
and	O
IL-13	B-protein
,	O
and	O
in	O
stimulated	O
human	B-cell_type
B	I-cell_type
cells	I-cell_type
,	O
on	O
which	O
IL-13R	B-protein
alpha	I-protein
1	I-protein
was	O
highly	O
expressed	O
,	O
IL-4	B-protein
and	O
IL-13	B-protein
induced	O
STAT3	B-protein
activation	O
.	O

Replacement	O
of	O
the	O
two	O
tyrosine	O
residues	O
completely	O
abolished	O
STAT3	B-protein
activation	O
,	O
although	O
replacing	O
either	O
tyrosine	O
residue	O
alone	O
retained	O
it	O
.	O

Furthermore	O
,	O
we	O
found	O
that	O
the	O
Box1	B-protein
region	I-protein
and	O
the	O
C-terminal	B-protein
tail	I-protein
of	O
IL-13R	B-protein
alpha	I-protein
1	I-protein
were	O
critical	O
for	O
binding	O
to	O
Tyk2	B-protein
,	O
and	O
activation	O
of	O
Jak1	B-protein
,	O
Tyk2	B-protein
,	O
the	O
insulin	B-protein
receptor	I-protein
substrate-1	I-protein
and	O
STAT6	B-protein
respectively	O
.	O

These	O
results	O
suggest	O
that	O
STAT3	B-protein
activation	O
is	O
involved	O
with	O
IL-4	B-protein
and	O
IL-13	B-protein
signals	O
in	O
human	B-cell_type
B	I-cell_type
cells	I-cell_type
along	O
with	O
the	O
activation	O
of	O
STAT6	B-protein
,	O
and	O
that	O
there	O
is	O
a	O
unique	O
sequence	O
in	O
IL-13R	B-protein
alpha	I-protein
1	I-protein
to	O
activate	O
STAT3	B-protein
.	O

-DOCSTART-	O

Functional	O
uncoupling	O
of	O
the	O
Janus	O
kinase	O
3-Stat5	O
pathway	O
in	O
malignant	O
growth	O
of	O
human	O
T	O
cell	O
leukemia	O
virus	O
type	O
1-transformed	O
human	B-cell_type
T	I-cell_type
cells	I-cell_type
.	O

Human	O
T	O
cell	O
leukemia	O
virus	O
type	O
1	O
(	O
HTLV-1	O
)	O
transforms	O
cytokine	B-protein
-dependent	O
T	B-cell_type
lymphocytes	I-cell_type
and	O
causes	O
adult	O
T	O
cell	O
leukemia	O
.	O

Janus	B-protein
tyrosine	I-protein
kinase	I-protein
(	I-protein
Jak	I-protein
)	I-protein
3	I-protein
and	O
transcription	B-protein
factors	I-protein
Stat5a	B-protein
and	O
Stat5b	B-protein
are	O
essential	O
for	O
the	O
proliferation	O
of	O
normal	B-cell_type
T	I-cell_type
cells	I-cell_type
and	O
are	O
constitutively	O
hyperactivated	O
in	O
both	O
HTLV-1-transformed	B-cell_line
human	I-cell_line
T	I-cell_line
cell	I-cell_line
lines	I-cell_line
and	O
lymphocytes	B-cell_type
isolated	O
from	O
HTLV-1-infected	O
patients	O
;	O
therefore	O
,	O
a	O
critical	O
role	O
for	O
the	O
Jak3-Stat5	O
pathway	O
in	O
the	O
progression	O
of	O
this	O
disease	O
has	O
been	O
postulated	O
.	O

We	O
recently	O
reported	O
that	O
tyrphostin	O
AG-490	O
selectively	O
blocked	O
IL-2	O
activation	O
of	O
Jak3/Stat5	B-protein
and	O
growth	O
of	O
murine	B-cell_line
T	I-cell_line
cell	I-cell_line
lines	I-cell_line
.	O

Here	O
we	O
demonstrate	O
that	O
disruption	O
of	O
Jak3/Stat5a/b	O
signaling	O
with	O
AG-490	O
(	O
50	O
&	O
mgr	O
;	O
M	O
)	O
blocked	O
the	O
proliferation	O
of	O
primary	B-cell_type
human	I-cell_type
T	I-cell_type
lymphocytes	I-cell_type
,	O
but	O
paradoxically	O
failed	O
to	O
inhibit	O
the	O
proliferation	O
of	O
HTLV-1-transformed	B-cell_line
human	I-cell_line
T	I-cell_line
cell	I-cell_line
lines	I-cell_line
,	O
HuT-102	B-cell_line
and	O
MT-2	B-cell_line
.	O

Structural	O
homologues	O
of	O
AG-490	O
also	O
inhibited	O
the	O
proliferation	O
of	O
primary	B-cell_type
human	I-cell_type
T	I-cell_type
cells	I-cell_type
,	O
but	O
not	O
HTLV-1-infected	B-cell_type
cells	I-cell_type
.	O

Disruption	O
of	O
constitutive	O
Jak3/Stat5	O
activation	O
by	O
AG-490	O
was	O
demonstrated	O
by	O
inhibition	O
of	O
1	O
)	O
tyrosine	O
phosphorylation	O
of	O
Jak3	B-protein
,	O
Stat5a	B-protein
(	O
Tyr	O
(	O
694	O
)	O
)	O
,	O
and	O
Stat5b	B-protein
(	O
Tyr	O
(	O
699	O
)	O
)	O
;	O
2	O
)	O
serine	O
phosphorylation	O
of	O
Stat5a	B-protein
(	O
Ser	O
(	O
726	O
)	O
)	O
as	O
determined	O
by	O
a	O
novel	O
phosphospecific	B-protein
Ab	I-protein
;	O
and	O
3	O
)	O
Stat5a/b	O
DNA	O
binding	O
to	O
the	O
Stat5-responsive	B-DNA
beta-casein	I-DNA
promoter	I-DNA
.	O

In	O
contrast	O
,	O
AG-490	O
had	O
no	O
effect	O
on	O
DNA	O
binding	O
by	O
p50/p65	B-protein
components	I-protein
of	O
NF-kappaB	B-protein
,	O
a	O
transcription	B-protein
factor	I-protein
activated	O
by	O
the	O
HTLV-1-encoded	B-protein
phosphoprotein	I-protein
,	I-protein
Tax	I-protein
.	O

Collectively	O
,	O
these	O
data	O
suggest	O
that	O
the	O
Jak3-Stat5	O
pathway	O
in	O
HTLV-1-transformed	B-cell_type
T	I-cell_type
cells	I-cell_type
has	O
become	O
functionally	O
redundant	O
for	O
proliferation	O
.	O

Reversal	O
of	O
this	O
functional	O
uncoupling	O
may	O
be	O
required	O
before	O
Jak3/Stat5	O
inhibitors	O
will	O
be	O
useful	O
in	O
the	O
treatment	O
of	O
this	O
malignancy	O
.	O

-DOCSTART-	O

The	O
Epstein-Barr	B-DNA
virus	I-DNA
promoter	I-DNA
initiating	O
B-cell	O
transformation	O
is	O
activated	O
by	O
RFX	B-protein
proteins	I-protein
and	O
the	O
B-cell-specific	B-protein
activator	I-protein
protein	I-protein
BSAP/Pax5	I-protein
.	O

Epstein-Barr	O
virus	O
(	O
EBV	O
)	O
-induced	O
B-cell	B-cell_type
growth	O
transformation	O
,	O
a	O
central	O
feature	O
of	O
the	O
virus	O
'	O
strategy	O
for	O
colonizing	O
the	O
human	O
B-cell	O
system	O
,	O
requires	O
full	O
virus	O
latent	O
gene	O
expression	O
and	O
is	O
initiated	O
by	O
transcription	O
from	O
the	O
viral	B-DNA
promoter	I-DNA
Wp	I-DNA
.	O

Interestingly	O
,	O
when	O
EBV	O
accesses	O
other	O
cell	O
types	O
,	O
this	O
growth-transforming	O
program	O
is	O
not	O
activated	O
.	O

The	O
present	O
work	O
focuses	O
on	O
a	O
region	O
of	O
Wp	B-DNA
which	O
in	O
reporter	O
assays	O
confers	O
B-cell	B-cell_type
-specific	O
activity	O
.	O

Bandshift	O
studies	O
indicate	O
that	O
this	O
region	O
contains	O
three	O
factor	O
binding	O
sites	O
,	O
termed	O
sites	B-DNA
B	I-DNA
,	I-DNA
C	I-DNA
,	I-DNA
and	I-DNA
D	I-DNA
,	O
in	O
addition	O
to	O
a	O
previously	O
characterized	O
CREB	B-DNA
site	I-DNA
.	O

Here	O
we	O
show	O
that	O
site	B-DNA
C	I-DNA
binds	O
members	O
of	O
the	O
ubiquitously	O
expressed	O
RFX	B-protein
family	I-protein
of	I-protein
proteins	I-protein
,	O
notably	O
RFX1	B-protein
,	O
RFX3	B-protein
,	O
and	O
the	O
associated	O
factor	O
MIBP1	B-protein
,	O
whereas	O
sites	B-DNA
B	I-DNA
and	I-DNA
D	I-DNA
both	O
bind	O
the	O
B-cell-specific	B-protein
activator	I-protein
protein	I-protein
BSAP/Pax5	I-protein
.	O

In	O
reporter	O
assays	O
with	O
mutant	B-DNA
Wp	I-DNA
constructs	I-DNA
,	O
the	O
loss	O
of	O
factor	O
binding	O
to	O
any	O
one	O
of	O
these	O
sites	O
severely	O
impaired	O
promoter	O
activity	O
in	O
B	B-cell_type
cells	I-cell_type
,	O
while	O
the	O
wild-type	B-DNA
promoter	I-DNA
could	O
be	O
activated	O
in	O
non-B	B-cell_type
cells	I-cell_type
by	O
ectopic	O
BSAP	O
expression	O
.	O

We	O
suggest	O
that	O
Wp	B-DNA
regulation	O
by	O
BSAP	B-protein
helps	O
to	O
ensure	O
the	O
B-cell	B-cell_type
specificity	O
of	O
EBV	O
's	O
growth-transforming	O
function	O
.	O

-DOCSTART-	O

Activation	O
of	O
the	O
Lck	B-protein
tyrosine	I-protein
protein	I-protein
kinase	I-protein
by	O
the	O
Herpesvirus	O
saimiri	O
tip	B-protein
protein	I-protein
involves	O
two	O
binding	O
interactions	O
.	O

The	O
Tip	B-protein
protein	I-protein
of	O
Herpesvirus	O
saimiri	O
strain	O
484C	O
binds	O
to	O
and	O
activates	O
the	O
Lck	B-protein
tyrosine	I-protein
protein	I-protein
kinase	I-protein
.	O

Two	O
sequences	O
in	O
the	O
Tip	B-protein
protein	I-protein
were	O
previously	O
shown	O
to	O
be	O
involved	O
in	O
binding	O
to	O
Lck	B-protein
.	O

A	O
proline-rich	B-protein
region	I-protein
,	O
residues	O
132-141	O
,	O
binds	O
to	O
the	O
SH3	B-protein
domain	I-protein
of	O
the	O
Lck	B-protein
protein	I-protein
.	O

We	O
show	O
here	O
that	O
the	O
other	O
Lck-binding	B-protein
domain	I-protein
,	O
residues	O
104-113	O
,	O
binds	O
to	O
the	O
carboxyl-terminal	B-protein
half	I-protein
of	O
Lck	B-protein
and	O
that	O
this	O
binding	O
does	O
not	O
require	O
the	O
Lck	B-protein
SH3	I-protein
domain	I-protein
.	O

Mutated	O
Tip	B-protein
containing	O
only	O
one	O
functional	O
Lck-binding	B-protein
domain	I-protein
can	O
bind	O
stably	O
to	O
Lck	B-protein
,	O
although	O
not	O
as	O
strongly	O
as	O
wild-type	B-protein
Tip	I-protein
.	O

Interaction	O
of	O
Tip	B-protein
with	O
Lck	B-protein
through	O
either	O
Lck-binding	B-protein
domain	I-protein
increases	O
the	O
activity	O
of	O
Lck	B-protein
in	O
vivo	O
.	O

Simultaneous	O
binding	O
of	O
both	O
domains	O
is	O
required	O
for	O
maximal	O
activation	O
of	O
Lck	B-protein
.	O

The	O
transient	O
expression	O
of	O
Tip	B-protein
in	O
T	B-cell_type
cells	I-cell_type
was	O
found	O
to	O
stimulate	O
both	O
Stat3	B-protein
-dependent	O
and	O
NF-AT	B-protein
-dependent	O
transcription	O
.	O

Mutant	O
forms	O
of	O
Tip	B-protein
lacking	O
one	O
or	O
the	O
other	O
of	O
the	O
two	O
Lck-binding	B-protein
domains	I-protein
retained	O
the	O
ability	O
to	O
stimulate	O
Stat3	B-protein
-dependent	O
transcription	O
.	O

Tip	B-protein
lacking	O
the	O
proline-rich	O
Lck-binding	B-protein
domain	I-protein
exhibited	O
almost	O
wild-type	O
activity	O
in	O
this	O
assay	O
.	O

In	O
contrast	O
,	O
ablation	O
of	O
either	O
Lck-binding	B-protein
domain	I-protein
abolished	O
the	O
ability	O
of	O
Tip	B-protein
to	O
stimulate	O
NF-AT	B-protein
-dependent	O
transcription	O
.	O

Full	O
biological	O
activity	O
of	O
Tip	B-protein
,	O
therefore	O
,	O
appears	O
to	O
require	O
both	O
Lck-binding	B-protein
domains	I-protein
.	O

-DOCSTART-	O

Accumulation	O
of	O
RXR	B-protein
alpha	I-protein
during	O
activation	O
of	O
cycling	O
human	B-cell_type
T	I-cell_type
lymphocytes	I-cell_type
:	O
modulation	O
of	O
RXRE	O
transactivation	O
function	O
by	O
mitogen-activated	O
protein	O
kinase	O
pathways	O
.	O

We	O
have	O
previously	O
reported	O
that	O
the	O
activation	O
of	O
resting	O
human	B-cell_type
immature	I-cell_type
peripheral	I-cell_type
blood	I-cell_type
T	I-cell_type
(	I-cell_type
PBT	I-cell_type
)	I-cell_type
lymphocytes	I-cell_type
is	O
associated	O
with	O
the	O
loss	O
of	O
retinoid	B-protein
X	I-protein
receptor	I-protein
alpha	I-protein
(	O
RXRalpha	B-protein
)	O
expression	O
.	O

In	O
the	O
present	O
study	O
,	O
we	O
have	O
demonstrated	O
that	O
,	O
unlike	O
resting	O
cells	O
,	O
activation	O
of	O
cycling	B-cell_type
human	I-cell_type
mature	I-cell_type
PBT	I-cell_type
lymphocytes	I-cell_type
,	O
and	O
T	B-cell_line
lymphocyte	I-cell_line
leukemia	I-cell_line
cell	I-cell_line
lines	I-cell_line
is	O
accompanied	O
by	O
the	O
accumulation	O
of	O
RXRalpha	B-protein
mRNA	O
and	O
protein	O
.	O

Interestingly	O
,	O
cyclosporin	O
A	O
further	O
augmented	O
RXRalpha	B-protein
expression	O
,	O
indicating	O
the	O
involvement	O
of	O
calcineurin	O
pathways	O
in	O
the	O
process	O
.	O

9-cis	O
retinoic	O
acid	O
inhibited	O
the	O
accumulation	O
,	O
suggesting	O
that	O
retinoids	O
can	O
regulate	O
the	O
synthesis	O
of	O
their	O
own	O
receptors	O
during	O
T	O
cell	O
activation	O
.	O

Transfection	O
analysis	O
in	O
Jurkat	B-cell_line
cells	I-cell_line
,	O
using	O
RXRE-dependent	O
reporter	O
assays	O
,	O
showed	O
that	O
RXRalpha	B-protein
accumulated	O
during	O
T	O
cell	O
activation	O
was	O
transcriptionally	O
inactive	O
.	O

To	O
investigate	O
the	O
mechanism	O
of	O
such	O
inhibition	O
,	O
the	O
role	O
of	O
two	O
mitogen-activated	O
protein	O
kinase	O
pathways	O
,	O
c-Jun	B-protein
N-terminal	I-protein
kinase	I-protein
(	O
JNK	B-protein
)	O
and	O
extracellular	B-protein
signal-regulated	I-protein
kinase	I-protein
(	O
ERK	B-protein
)	O
,	O
in	O
modulating	O
RXRE-dependent	O
transcription	O
,	O
was	O
explored	O
.	O

The	O
expression	O
of	O
constitutively	O
active	O
MAP/ERK	B-protein
kinase	I-protein
kinase	I-protein
1	I-protein
(	O
MEKK1	B-protein
)	O
inhibited	O
RXRE-dependent	O
transcription	O
,	O
whereas	O
dominant	B-protein
negative	I-protein
MEKK1	I-protein
increased	O
the	O
transcription	O
,	O
indicating	O
the	O
involvement	O
of	O
JNK	B-protein
signaling	O
pathways	O
in	O
the	O
process	O
.	O

In	O
contrast	O
,	O
expression	O
of	O
constitutively	B-protein
active	I-protein
MEK1	I-protein
,	O
which	O
activates	O
ERK	B-protein
pathway	O
,	O
enhanced	O
RXRE-dependent	O
activation	O
.	O

When	O
both	O
were	O
activated	O
simultaneously	O
,	O
JNK	B-protein
pathway	O
was	O
dominant	O
over	O
ERK	B-protein
pathway	O
and	O
resulted	O
in	O
inhibition	O
of	O
RXRE-mediated	O
transcription	O
.	O

These	O
data	O
demonstrate	O
a	O
dual	O
regulatory	O
control	O
of	O
RXRalpha	B-protein
expression	O
during	O
the	O
activation	O
of	O
resting	B-cell_type
and	I-cell_type
cycling	I-cell_type
T	I-cell_type
lymphocytes	I-cell_type
and	O
indicate	O
a	O
dynamic	O
balance	O
between	O
JNK	O
and	O
ERK	O
pathways	O
in	O
modulating	O
RXRE-mediated	O
transactivation	O
.	O

-DOCSTART-	O

The	O
proteasome	B-protein
regulates	O
receptor-mediated	O
endocytosis	O
of	O
interleukin-2	B-protein
.	O

Recent	O
studies	O
have	O
increasingly	O
implicated	O
the	O
proteasome	B-protein
in	O
the	O
regulation	O
of	O
cell	B-protein
surface	I-protein
receptors	I-protein
.	O

In	O
the	O
present	O
study	O
,	O
we	O
investigated	O
the	O
role	O
of	O
the	O
proteasome	B-protein
for	O
ligand-dependent	O
endocytosis	O
and	O
degradation	O
of	O
the	O
interleukin-2	B-protein
(	I-protein
IL-2	I-protein
)	I-protein
-interleukin-2	I-protein
receptor	I-protein
(	I-protein
IL-2R	I-protein
)	I-protein
complex	I-protein
.	O

Proteasome	O
inhibitors	O
impaired	O
internalization	O
of	O
IL-2.IL-2R	B-protein
and	O
prevented	O
the	O
lysosomal	O
degradation	O
of	O
this	O
cytokine	B-protein
.	O

Based	O
on	O
time-course	O
studies	O
,	O
proteasome	B-protein
activity	O
is	O
primarily	O
required	O
after	O
initial	O
endocytosis	O
of	O
the	O
IL-2.IL-2R	B-protein
.	O

Proteasome	O
function	O
was	O
also	O
necessary	O
for	O
the	O
lysosomal	O
degradation	O
of	O
IL-2	B-protein
internalized	O
by	O
IL-2R	B-protein
that	O
were	O
comprised	O
of	O
cytoplasmic	B-protein
tailless	I-protein
beta-	B-protein
or	I-protein
gamma	I-protein
c-subunits	I-protein
,	O
suggesting	O
that	O
the	O
target	O
protein	O
for	O
the	O
proteasome	B-protein
is	O
independent	O
of	O
either	O
the	O
cytoplasmic	B-protein
tail	I-protein
of	O
the	O
IL-2R	B-protein
beta-	B-protein
or	I-protein
gamma	I-protein
c-subunits	I-protein
and	O
their	O
associated	O
signaling	O
components	O
.	O

Therefore	O
,	O
a	O
functional	O
proteasome	B-protein
is	O
required	O
for	O
optimal	O
endocytosis	O
of	O
the	O
IL-2R/ligand	B-protein
complex	I-protein
and	O
is	O
essential	O
for	O
the	O
subsequent	O
lysosomal	O
degradation	O
of	O
IL-2	B-protein
,	O
possibly	O
by	O
regulating	O
trafficking	O
to	O
the	O
lysosome	O
.	O

-DOCSTART-	O

Functional	O
characterization	O
of	O
the	O
two	O
alternative	O
promoters	O
of	O
human	B-DNA
p45	I-DNA
NF-E2	I-DNA
gene	I-DNA
.	O

OBJECTIVE	O
:	O
The	O
transcription	B-protein
factor	I-protein
NF-E2	I-protein
,	O
a	O
heterodimeric	B-protein
protein	I-protein
complex	I-protein
composed	O
of	O
p45	B-protein
and	O
small	O
Maf	B-protein
family	I-protein
proteins	I-protein
,	O
is	O
considered	O
crucial	O
for	O
the	O
proper	O
differentiation	O
of	O
erythrocytes	B-cell_type
and	O
megakaryocytes	B-cell_type
in	O
vivo	O
.	O

We	O
report	O
the	O
results	O
of	O
studies	O
aimed	O
at	O
understanding	O
the	O
regulatory	O
mechanisms	O
controlling	O
p45	B-DNA
gene	I-DNA
expression	O
in	O
erythroid	B-cell_type
cells	I-cell_type
.	O

MATERIALS	O
AND	O
METHODS	O
:	O
Human	B-RNA
p45	I-RNA
mRNAs	I-RNA
have	O
two	O
alternative	O
isoforms	O
,	O
aNF-E2	B-RNA
and	O
fNF-E2	B-RNA
,	O
and	O
these	O
isoforms	O
are	O
transcribed	O
from	O
the	O
alternative	B-DNA
promoters	I-DNA
.	O

We	O
investigated	O
lineage-specific	O
expression	O
of	O
both	O
isomers	O
in	O
human	O
erythroid	B-cell_type
and	I-cell_type
megakaryocytic	I-cell_type
cells	I-cell_type
by	O
reverse	O
transcriptase	O
polymerase	O
chain	O
reaction	O
or	O
Northern	O
blot	O
analysis	O
.	O

For	O
functional	O
characterization	O
of	O
both	O
promoters	O
,	O
plasmids	O
in	O
which	O
reporter	B-DNA
genes	I-DNA
were	O
placed	O
under	O
the	O
control	O
of	O
a	O
series	O
of	O
truncated	O
or	O
mutated	O
promoter	B-DNA
fragments	I-DNA
were	O
transfected	O
to	O
human	B-cell_line
hematopoietic	I-cell_line
cell	I-cell_line
lines	I-cell_line
.	O

RESULTS	O
:	O
When	O
CD34	B-cell_type
(	I-cell_type
+	I-cell_type
)	I-cell_type
cells	I-cell_type
isolated	O
from	O
human	O
cord	O
blood	O
were	O
induced	O
to	O
unilineage	O
erythroid	O
or	O
megakaryocytic	O
differentiation	O
in	O
liquid	O
suspension	O
culture	O
,	O
both	O
transcripts	O
,	O
although	O
barely	O
detected	O
at	O
day	O
0	O
,	O
were	O
induced	O
in	O
both	O
erythroid	O
and	O
megakaryocytic	O
cultures	O
.	O

fNF-E2	B-RNA
mRNA	I-RNA
was	O
found	O
to	O
be	O
more	O
abundant	O
in	O
erythroid	B-cell_type
cells	I-cell_type
than	O
megakaryocytic	B-cell_type
cells	I-cell_type
at	O
day	O
7	O
of	O
culture	O
.	O

Although	O
both	O
isomers	O
were	O
expressed	O
in	O
human	B-cell_line
erythroid-megakaryocytic	I-cell_line
cell	I-cell_line
lines	I-cell_line
,	O
megakaryocytic	O
maturation	O
with	O
loss	O
of	O
erythroid	O
phenotype	O
induced	O
by	O
phorbol	O
12-myristate	O
13-acetate	O
(	O
PMA	O
)	O
resulted	O
in	O
exclusive	O
downregulation	O
of	O
fNF-E2	B-RNA
,	O
suggesting	O
that	O
fNF-E2	B-DNA
promoter	I-DNA
is	O
more	O
erythroid	O
specific	O
.	O

Functional	O
analysis	O
of	O
fNF-E2	B-DNA
promoter	I-DNA
showed	O
that	O
the	O
promoter	O
is	O
active	O
only	O
in	O
erythroid-megakaryocytic	B-cell_type
cells	I-cell_type
and	O
that	O
the	O
double	B-DNA
GATA	I-DNA
sit	I-DNA
e	O
in	O
the	O
proximal	O
region	O
is	O
necessary	O
for	O
its	O
efficient	O
activity	O
.	O

CONCLUSION	O
:	O
These	O
results	O
suggest	O
that	O
GATA	B-protein
proteins	I-protein
,	O
which	O
govern	O
the	O
differentiation	O
of	O
erythroid	B-cell_type
lineage	I-cell_type
cells	I-cell_type
,	O
are	O
required	O
for	O
full	O
promoter	O
activity	O
of	O
the	O
p45	B-DNA
gene	I-DNA
.	O

-DOCSTART-	O

Transcriptional	O
activation	O
of	O
heme	B-DNA
oxygenase-1	I-DNA
and	O
its	O
functional	O
significance	O
in	O
acetaminophen-induced	O
hepatitis	O
and	O
hepatocellular	O
injury	O
in	O
the	O
rat	O
.	O

BACKGROUND/AIM	O
:	O
Glutathione	O
depletion	O
contributes	O
to	O
acetaminophen	O
hepatotoxicity	O
and	O
is	O
known	O
to	O
induce	O
the	O
oxidative	O
stress	O
reactant	O
heme	B-protein
oxygenase-1	I-protein
.	O

The	O
metabolites	O
of	O
the	O
heme	O
oxygenase	O
pathway	O
,	O
biliverdin	O
,	O
carbon	O
monoxide	O
,	O
and	O
iron	O
may	O
modulate	O
acetaminophen	O
toxicity	O
.	O

The	O
aim	O
of	O
this	O
study	O
was	O
to	O
assess	O
cell-type	O
specific	O
expression	O
of	O
heme	B-DNA
oxygenase-1	I-DNA
and	O
its	O
impact	O
on	O
liver	O
injury	O
and	O
microcirculatory	O
disturbances	O
in	O
a	O
model	O
of	O
acetaminophen-induced	O
hepatitis	O
.	O

METHODS	O
:	O
Gene	O
expression	O
of	O
heme	B-DNA
oxygenase-1	I-DNA
was	O
studied	O
by	O
Northern-	O
and	O
Western	O
analysis	O
as	O
well	O
as	O
immunohistochemistry	O
.	O

The	O
time	O
course	O
of	O
heme	B-protein
oxygenase-1	I-protein
and	I-protein
-2	I-protein
,	O
cytokine-induced	B-protein
neutrophil	I-protein
chemoattractant-1	I-protein
,	O
and	O
intercellular	B-protein
adhesion	I-protein
molecule-1	I-protein
was	O
studied	O
by	O
Northern	O
analysis	O
.	O

DNA-binding	O
activity	O
of	O
nuclear	B-protein
factor-kappaB	I-protein
was	O
determined	O
by	O
electrophoretic	O
mobility	O
shift	O
assay	O
.	O

Sinusoidal	O
perfusion	O
and	O
leukocyte-endothelial	O
interactions	O
were	O
assessed	O
by	O
intravital	O
microscopy	O
.	O

RESULTS	O
:	O
Acetaminophen	O
caused	O
a	O
moderate	O
sinusoidal	O
perfusion	O
failure	O
(	O
-15	O
%	O
)	O
and	O
infiltration	O
of	O
neutrophils	B-cell_type
along	O
with	O
activation	O
of	O
nuclear	B-protein
factor-kappaB	I-protein
and	O
intercellular	B-protein
adhesion	I-protein
molecule-1	I-protein
and	O
cytokine-induced	B-protein
neutrophil	I-protein
chemoattractant-1	I-protein
mRNAs	O
.	O

Induction	O
of	O
heme	B-protein
oxygenase-1	I-protein
mRNA	O
and	O
protein	O
(	O
approximately	O
30-fold	O
)	O
in	O
hepatocytes	B-cell_type
and	O
non-parenchymal	B-cell_type
cells	I-cell_type
paralleled	O
the	O
inflammatory	O
response	O
.	O

Blockade	O
of	O
heme	O
oxygenase	O
activity	O
with	O
tin-protoporphyrin-IX	O
abrogated	O
acetaminophen-induced	O
hepatic	B-cell_type
neutrophil	I-cell_type
accumulation	O
and	O
nuclear	B-protein
factor-kappaB	I-protein
activation	O
,	O
but	O
failed	O
to	O
affect	O
sinusoidal	O
perfusion	O
and	O
liver	O
injury	O
.	O

CONCLUSIONS	O
:	O
The	O
inflammatory	O
response	O
associated	O
with	O
acetaminophen	O
hepatotoxicity	O
is	O
modulated	O
by	O
the	O
parallel	O
induction	O
of	O
the	O
heme	B-DNA
oxygenase-1	I-DNA
gene	I-DNA
.	O

However	O
,	O
heme	B-DNA
oxygenase-1	I-DNA
has	O
no	O
permissive	O
effect	O
on	O
sinusoidal	O
perfusion	O
and	O
does	O
not	O
affect	O
liver	O
injury	O
in	O
this	O
model	O
.	O

These	O
data	O
argue	O
against	O
a	O
central	O
role	O
of	O
nuclear	B-protein
factor-kappaB	I-protein
activation	O
and	O
neutrophil	O
infiltration	O
as	O
perpetuating	O
factors	O
of	O
liver	O
injury	O
in	O
acetaminophen	O
toxicity	O
.	O

-DOCSTART-	O

Tumor	B-protein
necrosis	I-protein
factor-alpha	I-protein
-induced	O
proliferation	O
requires	O
synthesis	O
of	O
granulocyte-macrophage	B-protein
colony-stimulating	I-protein
factor	I-protein
.	O

OBJECTIVE	O
:	O
Tumor	B-protein
necrosis	I-protein
factor-	I-protein
alpha	I-protein
(	O
TNF-alpha	B-protein
)	O
induces	O
a	O
variety	O
of	O
cellular	O
responses	O
,	O
some	O
of	O
them	O
being	O
at	O
least	O
seemingly	O
contradictory	O
.	O

Thus	O
,	O
we	O
set	O
out	O
to	O
find	O
differences	O
in	O
the	O
modes	O
of	O
proliferative	O
and	O
apoptotic	O
responses	O
to	O
TNF-	B-protein
alpha	I-protein
.	O

MATERIALS	O
AND	O
METHODS	O
:	O
We	O
screened	O
a	O
panel	O
of	O
acute	B-cell_line
myeloid	I-cell_line
leukemia-derived	I-cell_line
cell	I-cell_line
lines	I-cell_line
for	O
TNF-	B-protein
alpha	I-protein
-responsiveness	O
.	O

In	O
two	O
lines	O
(	O
OCI-AML-1	B-cell_line
,	O
OCI-AML-11	B-cell_line
)	O
,	O
TNF-	B-protein
alpha	I-protein
acted	O
as	O
an	O
apoptotic	O
agent	O
;	O
in	O
others	O
(	O
HU-3	B-cell_line
,	O
M-07e	B-cell_line
,	O
TF-1	B-cell_line
)	O
,	O
it	O
had	O
the	O
opposite	O
effect	O
,	O
preventing	O
apoptosis	O
and	O
inducing	O
proliferation	O
.	O

Direct	O
and	O
indirect	O
signaling	O
mechanisms	O
,	O
including	O
NF-kappaB	B-protein
activation	O
and	O
cytokine	O
synthesis	O
,	O
were	O
analyzed	O
.	O

RESULTS	O
:	O
All	O
cell	O
lines	O
tested	O
expressed	O
TNF-	B-protein
alpha	I-protein
receptors	I-protein
I	I-protein
and	I-protein
II	I-protein
and	O
responded	O
to	O
TNF-	B-protein
alpha	I-protein
by	O
upregulation	O
of	O
intercellular	B-protein
adhesion	I-protein
molecule-1	I-protein
.	O

In	O
contrast	O
to	O
granulocyte-macrophage	B-protein
colony-stimulating	I-protein
factor	I-protein
(	O
GM-CSF	B-protein
)	O
,	O
TNF-	B-protein
alpha	I-protein
did	O
not	O
activate	O
the	O
MAP	B-protein
kinase	I-protein
and	O
p70S6	O
kinase	O
pathways	O
.	O

Nevertheless	O
,	O
inhibitors	O
of	O
these	O
pathways	O
clearly	O
reduced	O
the	O
TNF-alpha	B-protein
-induced	O
cell	O
growth	O
,	O
indicating	O
that	O
TNF-	B-cell_type
alpha-proliferative	I-cell_type
cells	I-cell_type
produced	O
a	O
growth	O
factor	O
that	O
induced	O
proliferation	O
upon	O
stimulation	O
of	O
the	O
above	O
pathways	O
.	O

Anti-GM-CSF	B-protein
antibodies	I-protein
inhibited	O
the	O
TNF-alpha	B-protein
-induced	O
growth	O
,	O
suggesting	O
the	O
presence	O
of	O
an	O
autocrine	O
loop	O
for	O
cell	O
proliferation	O
mediated	O
by	O
GM-CSF	B-protein
.	O

Supporting	O
this	O
notion	O
,	O
TNF-alpha	B-protein
-induced	O
upregulation	O
of	O
GM-CSF	B-RNA
mRNA	I-RNA
levels	O
and	O
protein	O
secretion	O
in	O
the	O
TNF-alpha	B-protein
-proliferative	O
,	O
but	O
not	O
in	O
the	O
TNF-alpha-apoptotic	B-cell_line
cell	I-cell_line
lines	I-cell_line
.	O

CONCLUSION	O
:	O
These	O
data	O
identify	O
GM-CSF	B-protein
synthesis	O
as	O
an	O
early	O
and	O
essential	O
step	O
in	O
TNF-	O
alpha-induced	O
proliferation	O
.	O

We	O
show	O
for	O
the	O
first	O
time	O
that	O
TNF-alpha-treated	B-cell_line
cell	I-cell_line
lines	I-cell_line
producing	O
no	O
or	O
only	O
minimal	O
amounts	O
of	O
GM-CSF	B-protein
demonstrate	O
an	O
apoptotic	O
phenotype	O
,	O
while	O
cell	O
lines	O
with	O
high	O
GM-CSF	B-protein
expression	O
rates	O
can	O
escape	O
from	O
growth	O
arrest	O
or	O
even	O
apoptosis	O
.	O

In	O
this	O
context	O
,	O
we	O
discuss	O
arguments	O
pointing	O
at	O
NF-kappaB	B-protein
as	O
regulator	O
of	O
GM-CSF	B-protein
synthesis	O
and	O
thus	O
indirectly	O
as	O
regulator	O
for	O
the	O
escape	O
of	O
TNF-alpha	B-protein
-induced	O
apoptosis	O

-DOCSTART-	O

Glucocorticoid	B-protein
receptor	I-protein
content	O
of	O
T	B-cell_type
lymphocytes	I-cell_type
:	O
evidence	O
for	O
heterogeneity	O
.	O

Glucocorticoid	B-protein
receptors	I-protein
were	O
measured	O
in	O
T	B-cell_type
lymphocytes	I-cell_type
that	O
were	O
isolated	O
from	O
peripheral	O
blood	O
by	O
either	O
nylon	O
wool	O
filtration	O
or	O
E-rosette	O
sedimentation	O
.	O

T	B-cell_type
cells	I-cell_type
isolated	O
by	O
nylon	O
wool	O
filtration	O
specifically	O
bind	O
6.7	O
+/-	O
0.2	O
fmol	O
of	O
dexamethasone	O
per	O
million	O
cells	O
(	O
equivalent	O
to	O
4000	O
+/-	O
200	O
receptors	O
per	O
cell	O
)	O
,	O
whereas	O
T	B-cell_type
cells	I-cell_type
isolated	O
by	O
E-rosette	O
sedimentation	O
bind	O
12.0	O
+/-	O
0.7	O
fmol	O
of	O
dexamethasone	O
per	O
million	O
cells	O
(	O
equivalent	O
to	O
7200	O
+/-	O
400	O
receptors	O
per	O
cell	O
)	O
.	O

This	O
difference	O
in	O
the	O
amount	O
of	O
dexamethasone	O
bound	O
by	O
the	O
two	O
T	O
cell	O
preparations	O
was	O
significant	O
(	O
p	O
less	O
than	O
.001	O
)	O
and	O
was	O
present	O
immediately	O
after	O
cell	O
isolation	O
.	O

The	O
binding	O
affinities	O
of	O
the	O
different	O
T	B-cell_line
cell	I-cell_line
preparations	I-cell_line
for	O
dexamethasone	O
were	O
similar	O
.	O

T	O
cells	O
that	O
are	O
isolated	O
by	O
a	O
combination	O
of	O
nylon	O
wool	O
filtration	O
followed	O
by	O
E-rosette	O
sedimentation	O
bind	O
the	O
same	O
amount	O
of	O
dexamethasone	O
as	O
T	B-cell_type
cells	I-cell_type
isolated	O
by	O
nylon	O
wool	O
filtration	O
alone	O
.	O

T	B-cell_type
cells	I-cell_type
isolated	O
by	O
a	O
combination	O
of	O
E-rosette	O
sedimentation	O
following	O
by	O
nylon	O
wool	O
filtration	O
bind	O
less	O
dexamethasone	O
than	O
do	O
T	B-cell_type
cells	I-cell_type
isolated	O
by	O
E-rosette	O
sedimentation	O
alone	O
.	O

These	O
findings	O
suggest	O
that	O
T	B-cell_type
cells	I-cell_type
are	O
heterogeneous	O
with	O
respect	O
to	O
their	O
quantity	O
of	O
glucocorticoid	B-protein
receptors	I-protein
.	O

Isolation	O
of	O
T	B-cell_type
cells	I-cell_type
by	O
E-rosette	O
sedimentation	O
enriches	O
for	O
T	B-cell_type
cells	I-cell_type
that	O
have	O
a	O
greater	O
number	O
of	O
glucocorticoid	B-protein
receptors	I-protein
,	O
and	O
isolation	O
of	O
T	B-cell_type
cells	I-cell_type
by	O
nylon	O
wool	O
filtration	O
enriches	O
for	O
T	B-cell_type
cells	I-cell_type
that	O
have	O
a	O
lesser	O
number	O
of	O
glucocorticoid	B-protein
receptors	I-protein
.	O

-DOCSTART-	O

Glucocorticoid	B-protein
receptors	I-protein
and	O
glucocorticoid	O
sensitivity	O
of	O
human	B-cell_type
leukemic	I-cell_type
cells	I-cell_type
.	O

We	O
have	O
established	O
optimal	O
conditions	O
for	O
the	O
measurement	O
of	O
glucocorticoid	B-protein
receptors	I-protein
(	O
GR	B-protein
)	O
in	O
human	B-cell_type
white	I-cell_type
cells	I-cell_type
using	O
a	O
whole-cell	O
binding	O
assay	O
with	O
[	O
3H	O
]	O
dexamethasone	O
as	O
the	O
ligand	O
,	O
and	O
the	O
subsequent	O
determination	O
of	O
the	O
GR	B-protein
content	O
in	O
normal	B-cell_type
human	I-cell_type
lymphocytes	I-cell_type
and	O
in	O
leukemic	B-cell_type
cells	I-cell_type
of	O
patients	O
with	O
various	O
forms	O
of	O
acute	O
and	O
chronic	O
leukemia	O
.	O

A	O
number	O
of	O
leukemia	B-cell_line
cell	I-cell_line
lines	I-cell_line
in	O
continuous	O
culture	O
were	O
also	O
subjected	O
to	O
the	O
GR	B-protein
assay	O
,	O
and	O
the	O
results	O
were	O
correlated	O
with	O
the	O
sensitivity	O
of	O
these	O
cell	B-cell_line
lines	I-cell_line
to	O
glucocorticoid	O
steroids	O
in	O
vitro	O
.	O

The	O
GR	B-protein
content	O
of	O
normal	B-cell_type
human	I-cell_type
lymphocytes	I-cell_type
amounted	O
to	O
4	O
,	O
850	O
+/-	O
1	O
,	O
340	O
(	O
mean	O
+/-	O
SD	O
)	O
receptors/cell	O
.	O

The	O
mean	O
equilibrium	O
dissociation	O
constant	O
(	O
KD	O
)	O
of	O
the	O
interaction	O
of	O
[	O
3H	O
]	O
dexamethasone	O
with	O
the	O
GR	B-protein
was	O
1.2	O
x	O
10	O
(	O
-8	O
)	O
M	O
.	O

Steroidal	O
compounds	O
with	O
a	O
known	O
glucocorticoid	O
potency	O
effectively	O
competed	O
for	O
the	O
binding	O
,	O
whereas	O
steroids	O
devoid	O
of	O
glucocorticoid	O
activity	O
(	O
e.g.	O
estradiol-17	O
beta	O
and	O
testosterone	O
)	O
were	O
ineffective	O
.	O

The	O
GR	B-protein
content	O
of	O
the	O
blast	B-cell_type
cells	I-cell_type
obtained	O
from	O
eight	O
patients	O
suffering	O
from	O
acute	O
leukemia	O
and	O
four	O
patients	O
with	O
a	O
blast	O
crisis	O
of	O
chronic	O
myelocytic	O
leukemia	O
was	O
found	O
to	O
be	O
highly	O
variable	O
(	O
3	O
,	O
230-29	O
,	O
900	O
receptors/cell	O
)	O
,	O
while	O
the	O
lymphocytes	B-cell_type
of	O
six	O
patients	O
with	O
chronic	O
lymphatic	O
leukemia	O
contained	O
a	O
rather	O
stable	O
GR	B-protein
content	O
(	O
2	O
,	O
930-5	O
,	O
120	O
receptors/cell	O
)	O
,	O
which	O
was	O
comparable	O
with	O
that	O
of	O
normal	B-cell_type
lymphocytes	I-cell_type
.	O

GR	B-protein
was	O
identified	O
in	O
all	O
the	O
12	O
malignant	B-cell_line
continuous	I-cell_line
white	I-cell_line
cell	I-cell_line
lines	I-cell_line
studied	O
.	O

Large	O
cells	O
contained	O
more	O
GR	B-protein
than	O
the	O
smaller	O
ones	O
.	O

There	O
was	O
no	O
apparent	O
correlation	O
between	O
the	O
GR	B-protein
concentration	O
and	O
the	O
sensitivity	O
of	O
the	O
cells	O
in	O
vitro	O
to	O
glucocorticoids	O
as	O
judged	O
by	O
[	O
3H	O
]	O
thymidine	O
incorporation	O
studies	O
.	O

Distribution	O
of	O
the	O
surface	O
markers	O
in	O
the	O
leukemic	B-cell_line
cell	I-cell_line
lines	I-cell_line
did	O
not	O
relate	O
to	O
the	O
GR	B-protein
concentration	O
.	O

We	O
conclude	O
that	O
the	O
presence	O
of	O
GR	B-protein
is	O
probably	O
a	O
universal	O
feature	O
of	O
the	O
leukemic	B-cell_type
cells	I-cell_type
,	O
and	O
,	O
from	O
a	O
clinical	O
standpoint	O
,	O
probably	O
does	O
not	O
alone	O
imply	O
steroid	O
responsiveness	O
.	O

-DOCSTART-	O

Evidence	O
for	O
a	O
steroid	B-protein
receptor	I-protein
in	O
rheumatoid	B-cell_type
synovial	I-cell_type
tissue	I-cell_type
cells	I-cell_type
.	O

One	O
mechanism	O
by	O
which	O
glucocorticoids	O
could	O
exert	O
their	O
anti-inflammatory	O
action	O
is	O
via	O
rapidly	O
saturable	O
,	O
stereo-specific	B-protein
cytoplasmic	I-protein
protein	I-protein
receptors	I-protein
.	O

This	O
report	O
is	O
of	O
an	O
investigation	O
into	O
such	O
a	O
possibility	O
in	O
synovial	B-cell_type
cells	I-cell_type
.	O

Synovium	O
,	O
obtained	O
from	O
knee	O
joints	O
of	O
rheumatoid	O
patients	O
undergoing	O
surgery	O
,	O
was	O
incubated	O
with	O
clostridiopeptidase	B-protein
A	I-protein
and	O
trypsin-EDTA	B-protein
to	O
obtain	O
cell	O
suspensions	O
.	O

These	O
,	O
together	O
with	O
cells	O
obtained	O
from	O
synovial	O
fluid	O
aspirated	O
from	O
patients	O
with	O
rheumatoid	O
arthritis	O
,	O
were	O
identified	O
by	O
electron	O
microscopy	O
.	O

Duplicate	O
samples	O
of	O
these	O
cell	O
suspensions	O
were	O
incubated	O
with	O
increasing	O
concentrations	O
of	O
H3Dexamethasone	O
(	O
1	O
x	O
10	O
(	O
-10	O
)	O
M-1	O
x	O
10	O
(	O
-9	O
)	O
M	O
)	O
for	O
30	O
minutes	O
at	O
37	O
degrees	O
C	O
.	O

Analysis	O
of	O
the	O
proportion	O
of	O
steroid	O
bound	O
to	O
whole	B-cell_type
cells	I-cell_type
showed	O
evidence	O
for	O
specific	O
,	O
rapidly	O
saturable	O
,	O
receptors	O
in	O
the	O
cells	O
obtained	O
from	O
synovial	O
tissue	O
,	O
but	O
this	O
was	O
not	O
found	O
in	O
synovial	B-cell_type
fluid	I-cell_type
cells	I-cell_type
.	O

Electron	O
micrographs	O
showed	O
that	O
cells	O
obtained	O
from	O
synovial	O
tissue	O
consisted	O
of	O
synovial	B-cell_type
fibroblast	I-cell_type
-	O
and	O
macrophage-types	B-cell_type
,	O
lymphocytes	B-cell_type
,	O
monocytes	B-cell_type
and	O
macrophages	B-cell_type
.	O

Polymorphonuclear	B-cell_type
leucocytes	I-cell_type
appeared	O
to	O
be	O
absent	O
.	O

However	O
,	O
in	O
synovial	B-cell_type
fluid	I-cell_type
cell	I-cell_type
type	I-cell_type
polymorphonuclear	I-cell_type
leucocytes	I-cell_type
were	O
the	O
predominant	O
cell	O
type	O
.	O

We	O
concluded	O
from	O
this	O
,	O
that	O
one	O
or	O
more	O
of	O
the	O
cell	O
types	O
present	O
in	O
synovial	O
tissue	O
contain	O
a	O
specific	B-protein
steroid	I-protein
receptor	I-protein
,	O
but	O
that	O
this	O
is	O
lacking	O
in	O
synovial	B-cell_type
fluid	I-cell_type
polymorphonuclear	I-cell_type
leucocytes	I-cell_type
.	O

-DOCSTART-	O

Clinical	O
implications	O
of	O
glucocorticoid	B-protein
receptors	I-protein
in	O
human	O
leukemia	O
.	O

Glucorticoid	B-protein
receptors	I-protein
were	O
studied	O
in	O
various	O
populations	O
of	O
normal	O
human	B-cell_type
peripheral	I-cell_type
blood	I-cell_type
lymphocytes	I-cell_type
and	O
leukemic	B-cell_type
lymphoblasts	I-cell_type
.	O

Normal	B-cell_type
lymphocytes	I-cell_type
contain	O
low	O
levels	O
of	O
glucocorticoid	B-protein
receptor	I-protein
(	O
approximately	O
2	O
,	O
500	O
sites/cell	O
)	O
which	O
are	O
identical	O
in	O
T-	O
and	O
non-T-fractions	O
.	O

Phytohemagglutinin	B-protein
treatment	O
increases	O
levels	O
about	O
3-fold	O
.	O

Leukemic	B-cell_type
lymphoblasts	I-cell_type
contain	O
larger	O
numbers	O
of	O
receptor	O
sites	O
.	O

Presence	O
of	O
receptor	O
is	O
correlated	O
with	O
in	O
vitro	O
sensitivitiy	O
to	O
glucocorticoids	O
and	O
in	O
vivo	O
response	O
to	O
therapy	O
.	O

Quantity	O
of	O
receptor	O
is	O
also	O
correlated	O
with	O
complete	O
remission	O
duration	O
independently	O
of	O
leukemic	B-cell_type
cell	I-cell_type
type	I-cell_type
(	O
T	O
or	O
null	O
)	O
,	O
initial	O
WBC	B-cell_type
,	O
or	O
age	O
of	O
patient	O
.	O

Quantitative	O
determination	O
of	O
glucocorticoid	B-protein
receptor	I-protein
levels	O
in	O
acute	O
lymphoblastic	O
leukemia	O
may	O
be	O
of	O
value	O
both	O
as	O
an	O
independent	O
prognostic	O
variable	O
and	O
in	O
suggesting	O
which	O
patients	O
should	O
receive	O
glucocorticoid	O
therapy	O
.	O

-DOCSTART-	O

Functional	O
and	O
physical	O
interaction	O
of	O
protein-tyrosine	B-protein
kinases	I-protein
Fyn	I-protein
and	I-protein
Csk	I-protein
in	O
the	O
T-cell	O
signaling	O
system	O
.	O

The	O
Src-like	B-protein
protein-tyrosine	I-protein
kinase	I-protein
Fyn	B-protein
is	O
associated	O
with	O
T-cell	B-protein
antigen	I-protein
receptor	I-protein
.	O

Transient	O
expression	O
of	O
actively	O
mutated	O
Fyn	B-protein
,	O
having	O
Phe-528	O
instead	O
of	O
Tyr-528	O
or	O
Thr-338	O
instead	O
of	O
Ile-338	O
,	O
in	O
Jurkat	B-cell_type
T-cells	I-cell_type
stimulated	B-DNA
the	I-DNA
serum	I-DNA
response	I-DNA
element	I-DNA
(	O
SRE	B-DNA
)	O
,	O
12-O-tetradecanoyl-phorbol-13-acetate	B-DNA
response	I-DNA
element	I-DNA
,	O
cyclic	B-DNA
AMP	I-DNA
response	I-DNA
element	I-DNA
,	O
and	O
c-fos	B-DNA
promoter	I-DNA
.	O

The	O
stimulation	O
of	O
SRE	B-DNA
was	O
particularly	O
prominent	O
not	O
only	O
with	O
active	O
Fyn	B-protein
but	O
also	O
with	O
normal	B-protein
(	I-protein
wild-type	I-protein
)	I-protein
Fyn	I-protein
.	O

SRE	B-DNA
was	O
also	O
stimulated	O
by	O
both	O
normal	B-protein
and	I-protein
active	I-protein
Lck	I-protein
.	O

Furthermore	O
,	O
normal	B-protein
and	I-protein
active	I-protein
Fyn	I-protein
stimulated	O
transcription	O
from	O
the	O
IL-2	B-DNA
gene	I-DNA
promoter	I-DNA
when	O
transfected	O
cells	O
were	O
stimulated	O
by	O
concanavalin	B-protein
A	I-protein
plus	O
12-O-tetradecanoylphorbol-13-acetate	O
.	O

Under	O
the	O
same	O
conditions	O
,	O
Lck	B-protein
did	O
not	O
stimulate	O
IL-2	B-DNA
promoter	I-DNA
unless	O
it	O
was	O
activated	O
by	O
mutation	O
.	O

Interestingly	O
,	O
a	O
mutant	B-protein
Fyn	I-protein
,	O
which	O
has	O
deletions	O
within	O
the	O
SH2	B-protein
region	I-protein
and	O
so	O
is	O
able	O
to	O
transform	O
chicken	B-cell_type
embryo	I-cell_type
fibroblasts	I-cell_type
,	O
did	O
not	O
stimulate	O
either	O
the	O
c-fos	B-DNA
or	I-DNA
IL-2	I-DNA
promoter	I-DNA
,	O
suggesting	O
the	O
importance	O
of	O
this	O
region	O
in	O
T-cell	B-cell_type
signaling	O
.	O

Csk	B-protein
,	O
which	O
phosphorylates	O
tyrosine	O
residues	O
in	O
the	O
negative	O
regulatory	O
sites	O
of	O
Src	B-protein
family	I-protein
kinases	I-protein
,	O
down-regulated	O
Fyn-	O
and	O
Lck-mediated	O
stimulation	O
of	O
the	O
serum	B-DNA
response	I-DNA
element	I-DNA
and	O
Fyn-mediated	O
enhancement	O
of	O
IL-2	B-DNA
promoter	I-DNA
activity	O
.	O

These	O
data	O
suggest	O
that	O
Fyn	B-protein
and	O
Lck	B-protein
,	O
whose	O
activities	O
are	O
regulated	O
by	O
Csk	B-protein
,	O
are	O
involved	O
in	O
different	O
phases	O
of	O
T-cell	B-cell_type
activation	O
.	O

-DOCSTART-	O

A	O
novel	O
NF-kappa	B-protein
B	I-protein
complex	I-protein
containing	O
p65	B-protein
homodimers	I-protein
:	O
implications	O
for	O
transcriptional	O
control	O
at	O
the	O
level	O
of	O
subunit	O
dimerization	O
.	O

The	O
predominant	O
inducible	O
form	O
of	O
the	O
NF-kappa	B-protein
B	I-protein
transcription	I-protein
factor	I-protein
is	O
a	O
heteromeric	B-protein
complex	I-protein
containing	O
two	O
Rel-related	B-protein
DNA-binding	I-protein
subunits	I-protein
,	O
termed	B-protein
p65	I-protein
and	O
p50	B-protein
.	O

Prior	O
transfection	O
studies	O
have	O
shown	O
that	O
when	O
these	O
p65	B-protein
and	I-protein
p50	I-protein
subunits	I-protein
are	O
expressed	O
independently	O
as	O
stable	B-protein
homodimers	I-protein
,	O
p65	B-protein
stimulates	O
kappa	O
B-directed	O
transcription	O
,	O
whereas	O
p50	B-protein
functions	O
as	O
a	O
kappa	O
B-specific	O
repressor	O
.	O

While	O
authentic	O
p50	B-protein
homodimers	I-protein
(	O
previously	O
termed	O
KBF1	B-protein
)	O
have	O
been	O
detected	O
in	O
nuclear	O
extracts	O
from	O
nontransfected	O
cells	O
,	O
experimental	O
evidence	O
supporting	O
the	O
existence	O
of	O
p65	B-protein
homodimers	I-protein
in	O
vivo	O
was	O
lacking	O
.	O

We	O
now	O
provide	O
direct	O
biochemical	O
evidence	O
for	O
the	O
presence	O
of	O
an	O
endogenous	O
pool	O
of	O
inducible	O
p65	B-protein
homodimers	I-protein
in	O
intact	O
human	B-cell_type
T	I-cell_type
cells	I-cell_type
.	O

As	O
with	O
the	O
prototypical	O
NF-kappa	B-protein
B	I-protein
p50-p65	I-protein
heterodimer	I-protein
,	O
this	O
novel	O
p65	B-protein
homodimeric	I-protein
form	I-protein
of	O
NF-kappa	B-protein
B	I-protein
is	O
functionally	O
sequestered	O
in	O
the	O
cytoplasm	O
but	O
rapidly	O
appears	O
in	O
the	O
nuclear	O
compartment	O
following	O
cellular	O
stimulation	O
.	O

Site-directed	O
mutagenesis	O
studies	O
indicate	O
that	O
the	O
homodimerization	O
function	O
of	O
p65	B-protein
is	O
dependent	O
upon	O
the	O
presence	O
of	O
cysteine	O
216	O
and	O
a	O
conserved	O
recognition	O
motif	O
for	O
protein	B-protein
kinase	I-protein
A	I-protein
(	O
RRPS	B-protein
;	O
amino	O
acids	O
273	O
to	O
276	O
)	O
,	O
both	O
of	O
which	O
reside	O
within	O
a	O
91-amino-acid	O
segment	O
of	O
the	O
Rel	B-protein
homology	I-protein
domain	I-protein
that	O
mediates	O
self-association	O
.	O

In	O
contrast	O
,	O
mutations	O
at	O
these	O
two	O
sites	O
do	O
not	O
affect	O
heterodimerization	O
of	O
p65	B-protein
with	O
p50	B-protein
or	O
its	O
functional	O
interaction	O
with	O
I	O
kappa	O
B	O
alpha	O
.	O

These	O
later	O
findings	O
indicate	O
that	O
neither	O
homo-	O
nor	O
heterodimer	O
formation	O
is	O
an	O
absolute	O
prerequisite	O
for	O
I	B-protein
kappa	I-protein
B	I-protein
alpha	I-protein
recognition	O
of	O
p65	B-protein
.	O

Taken	O
together	O
with	O
prior	O
in	O
vivo	O
transcription	O
studies	O
,	O
these	O
results	O
suggest	O
that	O
the	O
biological	O
activities	O
of	O
p65	B-protein
and	I-protein
p50	I-protein
homodimers	I-protein
are	O
independently	O
regulated	O
,	O
thereby	O
providing	O
an	O
integrated	O
and	O
flexible	O
control	O
mechanism	O
for	O
the	O
rapid	O
activation	O
and	O
repression	O
of	O
NF-kappa	B-protein
B	I-protein
/	O
Rel	B-protein
-directed	O
gene	O
expression	O
.	O

-DOCSTART-	O

Carrier	O
determination	O
for	O
X-linked	O
agammaglobulinemia	O
using	O
X	O
inactivation	O
analysis	O
of	O
purified	B-cell_type
B	I-cell_type
cells	I-cell_type
.	O

We	O
report	O
the	O
development	O
of	O
a	O
relatively	O
quick	O
and	O
simple	O
method	O
for	O
the	O
assessment	O
of	O
X	O
inactivation	O
status	O
for	O
carrier	O
determination	O
in	O
families	O
affected	O
by	O
X-linked	O
agammaglobulinemia	O
(	O
XLA	O
)	O
.	O

This	O
method	O
utilises	O
an	O
immunomagnetic	O
separation	O
technique	O
for	O
B	B-cell_type
cell	I-cell_type
purification	O
and	O
a	O
polymerase	O
chain	O
reaction	O
(	O
PCR	O
)	O
based	O
assay	O
for	O
the	O
determination	O
of	O
methylation	O
status	O
at	O
the	O
androgen	B-DNA
receptor	I-DNA
(	I-DNA
AR	I-DNA
)	I-DNA
gene	I-DNA
locus	I-DNA
to	O
assess	O
whether	O
X	O
inactivation	O
is	O
random	O
or	O
non-random	O
at	O
this	O
locus	O
.	O

We	O
report	O
the	O
results	O
we	O
have	O
obtained	O
using	O
this	O
assay	O
to	O
investigate	O
females	O
known	O
to	O
be	O
carriers	O
of	O
various	O
X-linked	O
immunodeficiency	O
disorders	O
.	O

In	O
addition	O
,	O
we	O
investigated	O
four	O
females	O
from	O
different	O
families	O
affected	O
by	O
XLA	O
,	O
two	O
of	O
whom	O
were	O
of	O
unknown	O
carrier	O
status	O
,	O
and	O
we	O
discuss	O
the	O
results	O
obtained	O
with	O
this	O
and	O
other	O
X-inactivation	O
assays	O
.	O

A	O
similar	O
assay	O
has	O
recently	O
been	O
described	O
by	O
Allen	O
et	O
al.	O
(	O
1992	O
)	O
and	O
applied	O
to	O
members	O
of	O
one	O
family	O
affected	O
by	O
XLA	O
.	O

-DOCSTART-	O

Effects	O
of	O
IL-4	B-protein
and	O
Fc	B-protein
gamma	I-protein
receptor	I-protein
II	I-protein
engagement	O
on	O
Egr-1	B-protein
expression	O
during	O
stimulation	O
of	O
B	B-cell_type
lymphocytes	I-cell_type
by	O
membrane	O
immunoglobulin	O
crosslinking	O
.	O

Egr-1	B-DNA
is	O
an	O
immediate	B-DNA
early	I-DNA
gene	I-DNA
that	O
is	O
rapidly	O
upregulated	O
in	O
response	O
to	O
mitogenic	O
signals	O
induced	O
by	O
antigen	O
receptor	O
crosslinking	O
on	O
murine	B-cell_type
B	I-cell_type
lymphocytes	I-cell_type
.	O

It	O
has	O
been	O
shown	O
that	O
levels	O
of	O
Egr-1	B-DNA
expression	O
are	O
closely	O
correlated	O
with	O
B	B-cell_type
cell	I-cell_type
proliferation	O
in	O
several	O
models	O
of	O
B	O
cell	O
activation	O
and	O
tolerance	O
.	O

We	O
compared	O
the	O
expression	O
of	O
Egr-1	B-DNA
during	O
B	O
cell	O
stimulation	O
with	O
Fab'2	B-protein
and	O
IgG	B-protein
anti-immunoglobulin	I-protein
(	O
anti-Ig	B-protein
)	O
,	O
since	O
it	O
is	O
known	O
that	O
Fab'2	B-protein
anti-Ig	I-protein
is	O
mitogenic	O
while	O
IgG	B-protein
anti-Ig	I-protein
is	O
not	O
,	O
owing	O
to	O
a	O
dominant	O
inhibitory	O
effect	O
of	O
crosslinking	O
the	O
B	B-protein
cell	I-protein
Fc	I-protein
gamma	I-protein
RII	I-protein
to	O
membrane	B-protein
Ig	I-protein
.	O

While	O
mitogenic	O
doses	O
of	O
Fab'2	B-protein
anti-Ig	I-protein
induce	O
large	O
and	O
rapid	O
increases	O
in	O
Egr-1	O
expression	O
,	O
IgG	B-protein
anti-Ig	I-protein
results	O
in	O
smaller	O
increases	O
in	O
Egr-1	B-RNA
mRNA	I-RNA
,	O
comparable	O
to	O
that	O
seen	O
with	O
submitogenic	O
concentrations	O
of	O
Fab'2	B-protein
anti-Ig	I-protein
.	O

However	O
,	O
the	O
correlation	O
between	O
Egr-1	O
expression	O
and	O
B	O
cell	O
proliferation	O
breaks	O
down	O
when	O
IL-4	B-protein
is	O
added	O
as	O
a	O
co-mitogen	O
to	O
induce	O
B	O
cell	O
proliferation	O
with	O
IgG	B-protein
anti-Ig	I-protein
or	O
submitogenic	O
concentrations	O
of	O
Fab'2	B-protein
anti-Ig	I-protein
.	O

No	O
corresponding	O
increases	O
in	O
Egr-1	B-RNA
mRNA	I-RNA
levels	O
are	O
observed	O
when	O
IL-4	B-protein
is	O
added	O
.	O

Therefore	O
,	O
IL-4	B-protein
overcomes	O
Fc	O
receptor-mediated	O
inhibition	O
of	O
B	B-cell_type
cell	I-cell_type
proliferation	O
without	O
affecting	O
inhibition	O
of	O
Egr-1	B-RNA
mRNA	I-RNA
induction	O
,	O
as	O
demonstrated	O
earlier	O
for	O
c-myc	B-RNA
mRNA	I-RNA
in	O
this	O
system	O
.	O

-DOCSTART-	O

Identification	O
of	O
a	O
novel	O
cyclosporin-sensitive	B-DNA
element	I-DNA
in	O
the	O
human	B-DNA
tumor	I-DNA
necrosis	I-DNA
factor	I-DNA
alpha	I-DNA
gene	I-DNA
promoter	I-DNA
.	O

Tumor	B-protein
necrosis	I-protein
factor	I-protein
alpha	I-protein
(	O
TNF-alpha	B-protein
)	O
,	O
a	O
cytokine	B-protein
with	O
pleiotropic	O
biological	O
effects	O
,	O
is	O
produced	O
by	O
a	O
variety	O
of	O
cell	O
types	O
in	O
response	O
to	O
induction	O
by	O
diverse	O
stimuli	O
.	O

In	O
this	O
paper	O
,	O
TNF-alpha	B-RNA
mRNA	I-RNA
is	O
shown	O
to	O
be	O
highly	O
induced	O
in	O
a	O
murine	B-cell_line
T	I-cell_line
cell	I-cell_line
clone	I-cell_line
by	O
stimulation	O
with	O
T	B-protein
cell	I-protein
receptor	I-protein
(	O
TCR	B-protein
)	O
ligands	O
or	O
by	O
calcium	O
ionophores	O
alone	O
.	O

Induction	O
is	O
rapid	O
,	O
does	O
not	O
require	O
de	O
novo	O
protein	O
synthesis	O
,	O
and	O
is	O
completely	O
blocked	O
by	O
the	O
immunosuppressant	O
cyclosporin	O
A	O
(	O
CsA	O
)	O
.	O

We	O
have	O
identified	O
a	O
human	B-DNA
TNF-alpha	I-DNA
promoter	I-DNA
element	I-DNA
,	O
kappa	B-DNA
3	I-DNA
,	O
which	O
plays	O
a	O
key	O
role	O
in	O
the	O
calcium-mediated	O
inducibility	O
and	O
CsA	O
sensitivity	O
of	O
the	O
gene	O
.	O

In	O
electrophoretic	O
mobility	O
shift	O
assays	O
,	O
an	O
oligonucleotide	O
containing	O
kappa	B-DNA
3	I-DNA
forms	O
two	O
DNA	O
protein	O
complexes	O
with	O
proteins	O
that	O
are	O
present	O
in	O
extracts	O
from	O
unstimulated	B-cell_type
T	I-cell_type
cells	I-cell_type
.	O

These	O
complexes	O
appear	O
in	O
nuclear	O
extracts	O
only	O
after	O
T	O
cell	O
stimulation	O
.	O

Induction	O
of	O
the	O
inducible	O
nuclear	B-protein
complexes	I-protein
is	O
rapid	O
,	O
independent	O
of	O
protein	O
synthesis	O
,	O
and	O
blocked	O
by	O
CsA	O
,	O
and	O
thus	O
,	O
exactly	O
parallels	O
the	O
induction	O
of	O
TNF-alpha	B-RNA
mRNA	I-RNA
by	O
TCR	O
ligands	O
or	O
by	O
calcium	O
ionophore	O
.	O

Our	O
studies	O
indicate	O
that	O
the	O
kappa	B-protein
3	I-protein
binding	I-protein
factor	I-protein
resembles	O
the	O
preexisting	O
component	O
of	O
nuclear	B-protein
factor	I-protein
of	O
activated	B-cell_type
T	I-cell_type
cells	I-cell_type
.	O

Thus	O
,	O
the	O
TNF-alpha	B-DNA
gene	I-DNA
is	O
an	O
immediate	O
early	O
gene	O
in	O
activated	O
T	B-cell_type
cells	I-cell_type
and	O
provides	O
a	O
new	O
model	O
system	O
in	O
which	O
to	O
study	O
CsA-sensitive	B-DNA
gene	I-DNA
induction	O
in	O
activated	B-cell_type
T	I-cell_type
cells	I-cell_type
.	O

-DOCSTART-	O

Differences	O
in	O
expression	O
of	O
transcription	B-protein
factor	I-protein
AP-1	I-protein
in	O
human	B-cell_line
promyelocytic	I-cell_line
HL-60	I-cell_line
cells	I-cell_line
during	O
differentiation	O
towards	O
macrophages	B-cell_type
versus	O
granulocytes	B-cell_type
.	O

Commitment	O
of	O
HL-60	B-cell_line
cells	I-cell_line
to	O
macrophage	O
or	O
granulocytic	O
differentiation	O
was	O
achieved	O
by	O
incubation	O
with	O
4	O
beta-phorbol	O
12-myristate	O
13-acetate	O
(	O
PMA	O
)	O
for	O
30-60	O
min	O
or	O
with	O
dimethyl	O
sulphoxide	O
(	O
DMSO	O
)	O
for	O
24	O
h	O
respectively	O
.	O

The	O
commitment	O
stage	O
towards	O
PMA-induced	O
macrophage	O
differentiation	O
was	O
associated	O
with	O
increases	O
in	O
jun	O
B	O
and	O
c-fos	O
mRNA	O
levels	O
,	O
as	O
well	O
as	O
with	O
an	O
increase	O
in	O
the	O
binding	O
activity	O
of	O
transcription	B-protein
factor	I-protein
AP-1	I-protein
.	O

Nevertheless	O
,	O
gel	O
retardation	O
analysis	O
indicated	O
that	O
the	O
AP-1	B-protein
activity	O
detected	O
in	O
untreated	O
cells	O
was	O
drastically	O
reduced	O
during	O
the	O
commitment	O
stage	O
of	O
DMSO-induced	O
HL-60	O
differentiation	O
towards	O
granulocytes	B-cell_type
.	O

When	O
HL-60	B-cell_line
cells	I-cell_line
were	O
treated	O
with	O
sodium	O
butyrate	O
,	O
which	O
induced	O
monocytic	O
differentiation	O
,	O
a	O
remarkable	O
increase	O
in	O
AP-1	O
binding	O
activity	O
was	O
detected	O
.	O

Treatment	O
of	O
HL-60	B-cell_line
cells	I-cell_line
with	O
1	O
alpha	O
,	O
25-dihydroxyvitamin	O
D3	O
,	O
another	O
monocytic	O
differentiation	O
agent	O
,	O
induced	O
a	O
weak	O
,	O
but	O
appreciable	O
,	O
increase	O
in	O
AP-1	B-protein
activity	O
.	O

Furthermore	O
,	O
addition	O
of	O
sodium	O
butyrate	O
or	O
1	O
alpha	O
,	O
25-dihydroxyvitamin	O
D3	O
to	O
HL-60	B-cell_line
cells	I-cell_line
induced	O
the	O
expression	O
of	O
c-fos	B-DNA
,	I-DNA
c-jun	I-DNA
,	I-DNA
jun	I-DNA
B	I-DNA
and	I-DNA
jun	I-DNA
D	I-DNA
proto-oncogenes	I-DNA
.	O

In	O
contrast	O
,	O
when	O
HL-60	B-cell_line
cells	I-cell_line
were	O
treated	O
with	O
retinoic	O
acid	O
,	O
a	O
granulocytic	O
differentiation	O
inducer	O
,	O
no	O
enhanced	O
AP-1	O
binding	O
activity	O
was	O
observed	O
,	O
and	O
only	O
a	O
weak	O
increase	O
in	O
jun	B-RNA
D	I-RNA
mRNA	I-RNA
level	O
was	O
detected	O
.	O

These	O
data	O
indicate	O
that	O
formation	O
of	O
AP-1	B-protein
is	O
not	O
required	O
for	O
the	O
induction	O
of	O
HL-60	B-cell_line
differentiation	O
towards	O
granulocytes	B-cell_type
,	O
whereas	O
induction	O
of	O
monocytic	O
differentiation	O
is	O
correlated	O
with	O
an	O
increase	O
in	O
AP-1	B-protein
activity	O
.	O

The	O
differential	O
expression	O
of	O
AP-1	B-protein
activity	O
may	O
be	O
critical	O
in	O
the	O
differentiation	O
of	O
HL-60	B-cell_line
cells	I-cell_line
towards	O
monocytic	B-cell_type
or	I-cell_type
granulocytic	I-cell_type
lineages	I-cell_type

-DOCSTART-	O

Glucocorticoid	B-protein
receptors	I-protein
and	O
sensitivity	O
in	O
leukemias	O
.	O

In	O
an	O
attempt	O
to	O
investigate	O
the	O
utility	O
of	O
glucocorticoid	B-protein
receptor	I-protein
determination	O
to	O
predict	O
clinical	O
responsiveness	O
in	O
human	O
leukemias	O
we	O
have	O
studied	O
glucocorticoid	B-protein
receptors	I-protein
in	O
the	O
leukemic	B-cell_type
cells	I-cell_type
from	O
46	O
patients	O
and	O
in	O
the	O
lymphocytes	B-cell_type
from	O
18	O
normal	O
donors	O
.	O

In	O
the	O
normal	B-cell_type
lymphocytes	I-cell_type
there	O
were	O
3	O
,	O
875	O
(	O
Median	O
)	O
specific	O
binding	O
sites	O
per	O
cell	O
.	O

The	O
blasts	B-cell_type
from	O
17	O
patients	O
with	O
ANLL	O
had	O
on	O
average	O
higher	O
levels	O
of	O
binding	B-protein
sites	I-protein
per	O
cell	O
(	O
Median	O
=	O
7	O
,	O
250	O
,	O
range	O
:	O
0	O
to	O
15	O
,	O
295	O
)	O
than	O
the	O
other	O
leukemias	O
.	O

Of	O
the	O
15	O
patients	O
with	O
CLL	O
,	O
six	O
had	O
received	O
glucocorticoid	O
treatment	O
for	O
3	O
to	O
5	O
years	O
.	O

Their	O
lymphocytes	B-cell_type
had	O
lower	O
number	O
of	O
receptors	O
(	O
Median	O
=	O
2	O
,	O
000	O
)	O
than	O
the	O
other	O
cases	O
which	O
were	O
newly	O
diagnosed	O
(	O
Median	O
=	O
4	O
,	O
500	O
)	O
.	O

Four	O
patients	O
had	O
ALL/AUL	O
,	O
three	O
patients	O
had	O
blast	O
crisis	O
as	O
terminal	O
phase	O
of	O
CML	O
,	O
and	O
seven	O
had	O
leukemic	O
Non-Hodgkin	O
lymphomas	O
(	O
Median	O
=	O
3	O
,	O
500	O
sites/cell	O
)	O
.	O

In	O
24	O
patients	O
we	O
have	O
also	O
studied	O
the	O
in	O
vitro	O
sensitivity	O
of	O
the	O
leukemic	B-cell_type
cells	I-cell_type
to	O
dexamethasone	O
.	O

There	O
was	O
no	O
marked	O
correlation	O
between	O
glucocorticoid	B-protein
receptor	I-protein
levels	O
and	O
in	O
vitro	O
sensitivity	O
.	O

An	O
attempt	O
to	O
correlate	O
receptor	O
levels	O
with	O
clinical	O
responsiveness	O
demonstrated	O
that	O
glucocorticoid	B-protein
receptor	I-protein
determination	O
might	O
be	O
of	O
value	O
in	O
patients	O
with	O
lymphoid	O
malignancies	O
but	O
probably	O
not	O
in	O
patients	O
with	O
other	O
leukemias	O
.	O

-DOCSTART-	O

'Activation-labile	B-protein
'	I-protein
glucocorticoid-receptor	I-protein
complexes	I-protein
of	O
a	O
steroid-resistant	O
variant	O
of	O
CEM-C7	B-cell_line
human	I-cell_line
lymphoid	I-cell_line
cells	I-cell_line
.	O

For	O
cytoplasmic	B-protein
glucocorticoid-receptor	I-protein
complexes	I-protein
to	O
enter	O
and	O
accumulate	O
in	O
the	O
nucleus	O
a	O
temperature-dependent	O
event	O
,	O
'activation	O
'	O
is	O
required	O
.	O

Activation	O
can	O
be	O
achieved	O
in	O
vitro	O
by	O
increased	O
ionic	O
strength	O
,	O
dilution	O
or	O
gel	O
filtration	O
and	O
is	O
manifested	O
by	O
an	O
increased	O
affinity	O
of	O
steroid-receptor	B-protein
complex	I-protein
for	O
DNA	B-DNA
and	O
an	O
altered	O
elution	O
profile	O
from	O
ion-exchange	O
resins	O
.	O

Munck	O
and	O
Foley	O
have	O
shown	O
that	O
activated	B-protein
complexes	I-protein
isolated	O
from	O
thymocytes	O
elute	O
from	O
DEAE-cellulose	O
in	O
a	O
manner	O
identical	O
to	O
complexes	O
activated	O
in	O
vitro	O
.	O

We	O
report	O
here	O
that	O
DEAE-cellulose	O
chromatography	O
of	O
steroid-receptor	B-protein
complexes	I-protein
from	O
CEM-C7	B-cell_line
,	O
a	O
cloned	B-cell_line
human	I-cell_line
leukaemic	I-cell_line
T-cell	I-cell_line
line	I-cell_line
sensitive	O
to	O
the	O
cytolytic	O
action	O
of	O
glucocorticoids	O
,	O
and	O
its	O
steroid-resistant	B-cell_line
subclone	I-cell_line
4R4	I-cell_line
demonstrated	O
that	O
steroid	B-protein
receptors	I-protein
of	O
clone	B-cell_line
4R4	I-cell_line
can	O
not	O
form	O
stable	O
activated	O
complexes	O
.	O

This	O
defines	O
a	O
new	O
defect	O
in	O
receptor	O
action	O
,	O
activation	O
lability	O
(	O
r+act1	O
)	O
,	O
which	O
is	O
unlike	O
either	O
the	O
r-	B-cell_line
,	I-cell_line
r+nt-	I-cell_line
,	I-cell_line
or	I-cell_line
r+nti	I-cell_line
phenotypes	I-cell_line
previously	O
described	O
for	O
mouse	B-cell_line
lymphoid	I-cell_line
variants	I-cell_line
.	O

-DOCSTART-	O

Granulocytes	B-cell_type
in	O
the	O
endometrium	O
of	O
post-partum	O
women	O
.	O

Endometrial	O
samples	O
of	O
women	O
at	O
various	O
stages	O
of	O
gonadal	O
activity	O
after	O
parturition	O
were	O
examined	O
for	O
the	O
presence	O
and	O
numbers	O
of	O
endometrial	B-cell_type
granulocytes	I-cell_type
.	O

Although	O
samples	O
at	O
all	O
the	O
stages	O
contained	O
significant	O
numbers	O
of	O
the	O
granulocytes	B-cell_type
(	O
i.e.	O
greater	O
than	O
7/high-power	O
field	O
)	O
,	O
the	O
100	O
%	O
values	O
for	O
late-proliferative	O
and	O
adaptation	O
hyperplasia	O
were	O
significantly	O
higher	O
than	O
the	O
values	O
for	O
the	O
resting	O
(	O
81.8	O
%	O
)	O
,	O
early	O
(	O
82.4	O
%	O
)	O
and	O
mid-	O
(	O
87.9	O
%	O
)	O
proliferative	O
and	O
secretory	O
(	O
83.3	O
%	O
)	O
phases	O
.	O

We	O
suggest	O
that	O
this	O
correlates	O
with	O
the	O
suggestion	O
that	O
the	O
granulocytes	B-cell_type
constitute	O
a	O
receptor	O
system	O
for	O
oestrogens	O
.	O

-DOCSTART-	O

Interaction	O
of	O
glucocorticoids	O
with	O
macrophages	B-cell_type
.	O

Identification	O
of	O
glucocorticoid	B-protein
receptors	I-protein
in	O
monocytes	B-cell_type
and	O
macrophages	B-cell_type
.	O

Glucocorticoid	O
binding	O
was	O
measured	O
in	O
resident	O
and	O
thioglycollate-elicited	B-cell_type
mouse	I-cell_type
peritoneal	I-cell_type
macrophages	I-cell_type
,	O
rabbit	B-cell_type
alveolar	I-cell_type
macrophages	I-cell_type
,	O
and	O
human	B-cell_type
monocytes	I-cell_type
.	O

Two	O
assays	O
of	O
binding	O
were	O
used	O
--	O
an	O
assay	O
with	O
intact	B-cell_type
cells	I-cell_type
in	O
suspension	O
or	O
monolayers	O
,	O
and	O
an	O
assay	O
of	O
cytosol	O
and	O
nuclear	O
forms	O
of	O
glucocorticoid	B-protein
receptors	I-protein
.	O

The	O
mononuclear	B-cell_type
phagocytes	I-cell_type
contained	O
approximately	O
equal	O
to	O
4	O
--	O
10	O
X	O
10	O
(	O
3	O
)	O
high	B-protein
affinity	I-protein
receptor	I-protein
sites	I-protein
per	O
cell	O
,	O
with	O
dissociation	O
constants	O
of	O
approximately	O
equal	O
to	O
2	O
--	O
8	O
nM	O
dexamethasone	O
.	O

The	O
binding	O
to	O
the	O
saturable	B-protein
sites	I-protein
was	O
specific	O
for	O
steroids	O
with	O
glucocorticoid	O
or	O
antiglucocorticoid	O
activity	O
.	O

Cortisol	O
,	O
corticosterone	O
,	O
and	O
progesterone	O
competed	O
with	O
dexamethasone	O
for	O
binding	O
,	O
whereas	O
estradiol	O
,	O
dihydrotestosterone	O
,	O
and	O
11-epicortisol	O
competed	O
very	O
little	O
.	O

Binding	O
of	O
dexamethasone	O
to	O
cytosol	O
and	O
nuclear	O
forms	O
of	O
the	O
receptor	B-protein
complex	I-protein
and	O
temperature-sensitive	O
translocation	O
of	O
cytosol	O
forms	O
to	O
nuclear	O
forms	O
were	O
shown	O
.	O

At	O
37	O
degrees	O
C	O
the	O
predominant	O
form	O
of	O
the	O
hormone-receptor	B-protein
complex	I-protein
was	O
nuclear	O
.	O

These	O
results	O
demonstrate	O
that	O
corticosteroids	O
interact	O
with	O
macrophages	B-cell_type
at	O
physiological	O
concentrations	O
.	O

-DOCSTART-	O

Nitric	O
oxide	O
signaling	O
:	O
a	O
possible	O
role	O
for	O
G	B-protein
proteins	I-protein
.	O

We	O
have	O
previously	O
reported	O
various	O
inductive	O
effects	O
of	O
nitric	O
oxide	O
on	O
human	B-cell_type
PBMC	I-cell_type
.	O

We	O
describe	O
a	O
novel	O
and	O
potentially	O
important	O
mechanism	O
of	O
nitric	O
oxide	O
signaling-through	O
direct	O
activation	O
of	O
guanine	B-protein
nucleotide-binding	I-protein
proteins	I-protein
(	O
G	B-protein
proteins	I-protein
)	O
.	O

We	O
have	O
found	O
that	O
nitric	O
oxide	O
treatment	O
of	O
membranes	O
isolated	O
from	O
fresh	B-cell_type
human	I-cell_type
PBMC	I-cell_type
enhances	O
the	O
ability	O
of	O
these	O
membranes	O
to	O
hydrolyze	O
[	O
gamma-32P	O
]	O
GTP	O
and	O
bind	O
[	O
gamma-35S	O
]	O
GTP	O
.	O

In	O
addition	O
,	O
treatment	O
of	O
whole	O
cells	O
with	O
nitric	O
oxide	O
yielded	O
membranes	O
with	O
enhanced	O
GTPase	O
activity	O
.	O

Furthermore	O
,	O
the	O
GTPase	O
activity	O
of	O
pure	O
,	O
recombinant	O
Gs	B-protein
alpha	I-protein
,	O
Gi	B-protein
alpha	I-protein
1	I-protein
,	O
and	O
p21ras	B-protein
was	O
greatly	O
enhanced	O
by	O
nitric	O
oxide	O
.	O

In	O
support	O
of	O
the	O
existence	O
of	O
this	O
pathway	O
in	O
whole	O
cells	O
,	O
we	O
found	O
that	O
the	O
G	O
protein	O
inhibitor	O
,	O
GDP-beta-S	O
,	O
blocked	O
NF-kappa	B-protein
B	I-protein
translocation	O
induced	O
by	O
nitric	O
oxide	O
or	O
LPS	O
in	O
permeabilized	B-cell_type
cells	I-cell_type
.	O

In	O
addition	O
,	O
nitric	O
oxide	O
greatly	O
reduced	O
the	O
pertussis	O
toxin-mediated	O
ADP-ribosylation	O
of	O
45-	B-protein
and	I-protein
41-kDa	I-protein
proteins	I-protein
in	O
membranes	O
of	O
these	O
cells	O
.	O

Because	O
G	B-protein
proteins	I-protein
play	O
a	O
central	O
role	O
in	O
many	O
diverse	O
signaling	O
systems	O
,	O
activation	O
by	O
an	O
endogenous	O
and	O
inducible	O
oxidant	O
may	O
represent	O
a	O
novel	O
signaling	O
pathway	O
.	O

-DOCSTART-	O

The	O
granulocyte-macrophage	B-DNA
colony-stimulating	I-DNA
factor	I-DNA
promoter	I-DNA
cis-acting	I-DNA
element	I-DNA
CLE0	B-DNA
mediates	O
induction	O
signals	O
in	O
T	B-cell_type
cells	I-cell_type
and	O
is	O
recognized	O
by	O
factors	O
related	O
to	O
AP1	B-protein
and	O
NFAT	B-protein
.	O

Expression	O
of	O
the	O
granulocyte-macrophage	B-DNA
colony-stimulating	I-DNA
factor	I-DNA
(	I-DNA
GM-CSF	I-DNA
)	I-DNA
gene	I-DNA
in	O
T	O
cells	O
is	O
activated	O
by	O
the	O
combination	O
of	O
phorbol	O
ester	O
(	O
phorbol	O
myristate	O
acetate	O
)	O
and	O
calcium	O
ionophore	O
(	O
A23187	O
)	O
,	O
which	O
mimic	O
antigen	O
stimulation	O
through	O
the	O
T-cell	B-protein
receptor	I-protein
.	O

We	O
have	O
previously	O
shown	O
that	O
a	O
fragment	O
containing	O
bp	B-DNA
-95	I-DNA
to	I-DNA
+27	I-DNA
of	O
the	O
mouse	B-DNA
GM-CSF	I-DNA
promoter	I-DNA
can	O
confer	O
inducibility	O
to	O
reporter	B-DNA
genes	I-DNA
in	O
the	O
human	B-cell_line
Jurkat	I-cell_line
T-cell	I-cell_line
line	I-cell_line
.	O

Here	O
we	O
use	O
an	O
in	O
vitro	O
transcription	O
system	O
to	O
demonstrate	O
that	O
a	O
cis-acting	B-DNA
element	I-DNA
(	O
positions	B-DNA
-54	I-DNA
to	I-DNA
-40	I-DNA
)	O
,	O
referred	O
to	O
as	O
CLE0	B-DNA
,	O
is	O
a	O
target	O
for	O
the	O
induction	O
signals	O
.	O

We	O
observed	O
induction	O
with	O
templates	O
containing	O
intact	O
CLE0	B-DNA
but	O
not	O
with	O
templates	O
with	O
deleted	B-DNA
or	I-DNA
mutated	I-DNA
CLE0	I-DNA
.	O

We	O
also	O
observed	O
that	O
two	O
distinct	O
signals	O
were	O
required	O
for	O
the	O
stimulation	O
through	O
CLE0	B-DNA
,	O
since	O
only	O
extracts	O
from	O
cells	O
treated	O
with	O
both	O
phorbol	O
myristate	O
acetate	O
and	O
A23187	O
supported	O
optimal	O
induction	O
.	O

Stimulation	O
probably	O
was	O
mediated	O
by	O
CLE0-binding	B-protein
proteins	I-protein
because	O
depletion	O
of	O
these	O
proteins	O
specifically	O
reduced	O
GM-CSF	B-protein
transcription	O
.	O

One	O
of	O
the	O
binding	B-protein
factors	I-protein
possessed	O
biochemical	O
and	O
immunological	O
features	O
identical	O
to	O
those	O
of	O
the	O
transcription	B-protein
factor	I-protein
AP1	I-protein
.	O

Another	O
factor	O
resembled	O
the	O
T-cell-specific	B-protein
factor	I-protein
NFAT	I-protein
.	O

The	O
characteristics	O
of	O
these	O
two	O
factors	O
are	O
consistent	O
with	O
their	O
involvement	O
in	O
GM-CSF	O
induction	O
.	O

The	O
presence	O
of	O
CLE0-like	B-DNA
elements	I-DNA
in	O
the	O
promoters	B-DNA
of	O
interleukin-3	B-DNA
(	I-DNA
IL-3	I-DNA
)	I-DNA
,	I-DNA
IL-4	I-DNA
,	I-DNA
IL-5	I-DNA
,	I-DNA
GM-CSF	I-DNA
,	I-DNA
and	I-DNA
NFAT	I-DNA
sites	I-DNA
in	O
the	O
IL-2	B-DNA
promoter	I-DNA
suggests	O
that	O
the	O
factors	O
we	O
detected	O
,	O
or	O
related	O
factors	O
that	O
recognize	O
these	O
sites	O
,	O
may	O
account	O
for	O
the	O
coordinate	O
induction	O
of	O
these	O
genes	O
during	O
T-cell	B-cell_type
activation	O
.	O

-DOCSTART-	O

Identification	O
and	O
characterization	O
of	O
an	O
Alu-containing	B-DNA
,	I-DNA
T-cell-specific	I-DNA
enhancer	I-DNA
located	O
in	O
the	O
last	O
intron	B-DNA
of	O
the	O
human	B-DNA
CD8	I-DNA
alpha	I-DNA
gene	I-DNA
.	O

Expression	O
of	O
the	O
human	B-DNA
CD8	I-DNA
alpha	I-DNA
gene	I-DNA
is	O
restricted	O
to	O
cells	O
of	O
the	O
lymphoid	B-cell_type
lineage	I-cell_type
and	O
developmentally	O
regulated	O
during	O
thymopoiesis	O
.	O

As	O
an	O
initial	O
step	O
towards	O
understanding	O
the	O
molecular	O
basis	O
for	O
tissue-specific	O
expression	O
of	O
this	O
gene	O
,	O
we	O
surveyed	O
the	O
surrounding	O
chromatin	O
structure	O
for	O
potential	O
cis-acting	B-DNA
regulatory	I-DNA
regions	I-DNA
by	O
DNase	B-protein
I	I-protein
hypersensitivity	O
mapping	O
and	O
found	O
four	O
hypersensitive	O
sites	O
,	O
three	O
of	O
which	O
were	O
T	O
cell	O
restricted	O
.	O

By	O
using	O
a	O
reporter-based	O
expression	O
approach	O
,	O
a	O
T-cell-specific	B-DNA
enhancer	I-DNA
was	O
identified	O
by	O
its	O
close	O
association	O
with	O
a	O
prominent	O
T-cell-restricted	B-DNA
hypersensitive	I-DNA
sites	I-DNA
in	O
the	O
last	O
intron	O
of	O
the	O
CD8	B-DNA
alpha	I-DNA
gene	I-DNA
.	O

Deletion	O
studies	O
demonstrated	O
that	O
the	O
minimal	O
enhancer	O
is	O
adjacent	O
to	O
a	O
negative	O
regulatory	O
element	O
.	O

DNA	O
sequence	O
analysis	O
of	O
the	O
minimal	B-DNA
enhancer	I-DNA
revealed	O
a	O
striking	O
cluster	O
of	O
consensus	B-DNA
binding	I-DNA
sites	I-DNA
for	O
Ets-1	B-protein
,	I-protein
TCF-1	I-protein
,	I-protein
CRE	I-protein
,	I-protein
GATA-3	I-protein
,	I-protein
LyF-1	I-protein
,	I-protein
and	I-protein
bHLH	I-protein
proteins	I-protein
which	O
were	O
verified	O
by	O
electrophoretic	O
mobility	O
shift	O
assays	O
.	O

In	O
addition	O
,	O
the	O
5	B-DNA
'	I-DNA
end	I-DNA
of	O
the	O
enhancer	B-DNA
was	O
composed	O
of	O
an	O
Alu	O
repeat	O
which	O
contained	O
the	O
GATA-3	B-DNA
,	I-DNA
bHLH	I-DNA
,	I-DNA
and	I-DNA
LyF-1	I-DNA
binding	I-DNA
sites	I-DNA
.	O

Site-directed	O
mutation	O
of	O
the	O
Ets-1	B-DNA
and	I-DNA
GATA-3	I-DNA
sites	I-DNA
dramatically	O
reduced	O
enhancer	O
activity	O
.	O

The	O
functional	O
importance	O
of	O
the	O
other	O
binding	O
sites	O
only	O
became	O
apparent	O
when	O
combinations	O
of	O
mutations	O
were	O
analyzed	O
.	O

Taken	O
together	O
,	O
these	O
results	O
suggest	O
that	O
the	O
human	B-DNA
CD8	I-DNA
alpha	I-DNA
gene	I-DNA
is	O
regulated	O
by	O
the	O
interaction	O
of	O
multiple	O
T-cell	B-protein
nuclear	I-protein
proteins	I-protein
with	O
a	O
transcriptional	O
enhancer	O
located	O
in	O
the	O
last	O
intron	B-DNA
of	O
the	O
gene	O
.	O

Comparison	O
of	O
the	O
CD8	B-DNA
alpha	I-DNA
enhancer	I-DNA
with	O
other	O
recently	O
identified	O
T-cell-specific	B-DNA
regulatory	I-DNA
elements	I-DNA
suggests	O
that	O
a	O
common	O
set	O
of	O
transcription	B-protein
factors	I-protein
regulates	O
several	O
T-cell	B-DNA
genes	I-DNA
.	O

-DOCSTART-	O

Molecular	O
regulation	O
of	O
the	O
human	B-DNA
IL-3	I-DNA
gene	I-DNA
:	O
inducible	O
T	O
cell-restricted	O
expression	O
requires	O
intact	O
AP-1	B-DNA
and	I-DNA
Elf-1	I-DNA
nuclear	I-DNA
protein	I-DNA
binding	I-DNA
sites	I-DNA
.	O

Interleukin	B-protein
3	I-protein
(	O
IL-3	B-protein
)	O
is	O
a	O
hematopoietic	B-protein
stem-cell	I-protein
growth	I-protein
and	I-protein
differentiation	I-protein
factor	I-protein
that	O
is	O
expressed	O
solely	O
in	O
activated	O
T	B-cell_type
and	I-cell_type
NK	I-cell_type
cells	I-cell_type
.	O

Studies	O
to	O
date	O
have	O
identified	O
elements	O
5	O
'	O
to	O
the	O
IL-3	B-DNA
coding	I-DNA
sequences	I-DNA
that	O
regulate	O
its	O
transcription	O
,	O
but	O
the	O
sequences	O
that	O
confer	O
T	O
cell-specific	O
expression	O
remain	O
to	O
be	O
clearly	O
defined	O
.	O

We	O
have	O
now	O
identified	O
DNA	B-DNA
sequences	I-DNA
that	O
are	O
required	O
for	O
T	O
cell-restricted	O
IL-3	B-DNA
gene	I-DNA
transcription	O
.	O

A	O
series	O
of	O
transient	O
transfections	O
performed	O
with	O
human	B-DNA
IL-3-chloramphenicol	I-DNA
acetyltransferase	I-DNA
(	I-DNA
CAT	I-DNA
)	I-DNA
reporter	I-DNA
plasmids	I-DNA
in	O
T	B-cell_type
and	I-cell_type
non-T	I-cell_type
cells	I-cell_type
revealed	O
that	O
a	O
plasmid	B-DNA
containing	O
319	O
bp	O
of	O
5	B-DNA
'	I-DNA
flanking	I-DNA
sequences	I-DNA
was	O
active	O
exclusively	O
in	O
T	B-cell_type
cells	I-cell_type
.	O

Deletion	O
analysis	O
revealed	O
that	O
T	B-cell_type
cell	I-cell_type
specificity	O
was	O
conferred	O
by	O
a	O
49-bp	B-DNA
fragment	I-DNA
(	O
bp	B-DNA
-319	I-DNA
to	I-DNA
-270	I-DNA
)	O
that	O
included	O
a	O
potential	B-DNA
binding	I-DNA
site	I-DNA
for	O
AP-1	B-protein
transcription	I-protein
factors	I-protein
6	O
bp	O
upstream	O
of	O
a	O
binding	B-DNA
site	I-DNA
for	I-DNA
Elf-1	I-DNA
,	O
a	O
member	O
of	O
the	O
Ets	B-protein
family	I-protein
of	O
transcription	B-protein
factors	I-protein
.	O

DNaseI	B-protein
footprint	O
and	O
electrophoretic	O
mobility	O
shift	O
assay	O
analyses	O
performed	O
with	O
MLA-144	B-cell_line
T	I-cell_line
cell	I-cell_line
nuclear	O
extracts	O
demonstrated	O
that	O
this	O
49-bp	B-DNA
region	I-DNA
contains	O
a	O
nuclear	B-DNA
protein	I-DNA
binding	I-DNA
region	I-DNA
that	O
includes	O
consensus	B-DNA
AP-1	I-DNA
and	I-DNA
Elf-1	I-DNA
binding	I-DNA
sites	I-DNA
.	O

In	O
addition	O
,	O
extracts	O
prepared	O
from	O
purified	B-cell_type
human	I-cell_type
T	I-cell_type
cells	I-cell_type
contained	O
proteins	O
that	O
bound	O
to	O
synthetic	O
oligonucleotides	O
corresponding	O
to	O
the	O
AP-1	B-DNA
and	I-DNA
Elf-1	I-DNA
binding	I-DNA
sites	I-DNA
.	O

In	O
vitro-transcribed	O
and	O
-translated	O
Elf-1	B-protein
protein	I-protein
bound	O
specifically	O
to	O
the	O
Elf-1	B-DNA
site	I-DNA
,	O
and	O
Elf-1	O
antisera	O
competed	O
and	O
super	O
shifted	O
nuclear	O
protein	O
complexes	O
present	O
in	O
MLA-144	B-cell_line
nuclear	O
extracts	O
.	O

Moreover	O
,	O
addition	O
of	O
anti-Jun	O
family	O
antiserum	O
in	O
electrophoretic	O
mobility	O
shift	O
assay	O
reactions	O
completely	O
blocked	O
formation	O
of	O
the	O
AP-1-related	B-protein
complexes	I-protein
.	O

Transient	O
transfection	O
studies	O
in	O
MLA-144	B-cell_line
T	I-cell_line
cells	I-cell_line
revealed	O
that	O
constructs	O
containing	O
mutations	O
in	O
the	O
AP-1	B-DNA
site	I-DNA
almost	O
completely	O
abolished	O
CAT	O
activity	O
while	O
mutation	O
of	O
the	O
Elf-1	B-DNA
site	I-DNA
or	O
the	O
NF-IL-3	B-DNA
site	I-DNA
,	O
a	O
previously	O
described	O
nuclear	B-DNA
protein	I-DNA
binding	I-DNA
site	I-DNA
(	O
bp.	B-DNA
-155	I-DNA
to	I-DNA
-148	I-DNA
)	O
in	O
the	O
IL-3	B-DNA
promoter	I-DNA
,	O
reduced	O
CAT	O
activity	O
to	O
<	O
25	O
%	O
of	O
the	O
activity	O
given	O
by	O
wild-type	O
constructs	O
.	O

We	O
conclude	O
that	O
expression	O
of	O
the	O
human	B-DNA
IL-3	I-DNA
gene	I-DNA
requires	O
the	O
AP-1	B-DNA
and	I-DNA
Elf-1	I-DNA
binding	I-DNA
sites	I-DNA
;	O
however	O
,	O
unlike	O
other	O
previously	O
characterized	O
cytokine	B-DNA
genes	I-DNA
such	O
as	O
IL-2	B-protein
,	O
the	O
AP-1	B-protein
and	I-protein
Elf-1	I-protein
factors	I-protein
can	O
bind	O
independently	O
in	O
the	O
IL-3	B-DNA
gene	I-DNA
.	O

(	O
ABSTRACT	O
TRUNCATED	O
AT	O
400	O
WORDS	O
)	O

-DOCSTART-	O

Combination	O
IL-2	B-protein
and	O
IL-4	B-protein
reduces	O
glucocorticoid	O
receptor-binding	O
affinity	O
and	O
T	B-cell_type
cell	I-cell_type
response	O
to	O
glucocorticoids	O
.	O

The	O
mechanisms	O
contributing	O
to	O
persistent	O
T	B-cell_type
cell	I-cell_type
activation	O
and	O
poor	O
response	O
to	O
glucocorticoids	O
in	O
chronic	O
inflammatory	O
illnesses	O
such	O
as	O
steroid	O
resistant	O
(	O
SR	O
)	O
asthma	O
are	O
poorly	O
defined	O
.	O

We	O
examined	O
the	O
possibility	O
that	O
certain	O
cytokines	O
,	O
specifically	O
IL-2	B-protein
and	O
IL-4	B-protein
,	O
could	O
affect	O
T	O
cell	O
response	O
to	O
glucocorticoids	O
.	O

A	O
[	O
3H	O
]	O
dexamethasone	O
radioligand-binding	O
assay	O
was	O
used	O
to	O
measure	O
the	O
number	O
of	O
glucocorticoid	B-protein
receptors	I-protein
(	O
GR	B-protein
)	O
and	O
dissociation	O
constant	O
(	O
Kd	O
)	O
in	O
PBMC	B-cell_type
from	O
normal	O
donors	O
and	O
patients	O
with	O
SR	O
asthma	O
,	O
cultured	O
in	O
the	O
absence	O
and	O
presence	O
of	O
these	O
cytokines	B-protein
.	O

PBMC	O
from	O
normal	O
donors	O
incubated	O
for	O
48	O
h	O
in	O
the	O
presence	O
of	O
combination	O
IL-2	B-protein
+	O
IL-4	B-protein
had	O
nuclear	O
GR	O
with	O
significantly	O
reduced	O
binding	O
affinity	O
(	O
GR	O
Kd	O
=	O
36.1	O
+/-	O
1.63	O
nM	O
,	O
mean	O
+/-	O
SEM	O
;	O
p	O
=	O
0.0001	O
)	O
as	O
compared	O
with	O
PBMC	B-cell_type
incubated	O
with	O
medium	O
alone	O
(	O
GR	O
Kd	O
=	O
6.74	O
+/-	O
0.46	O
nM	O
)	O
.	O

The	O
cytosolic	O
GR	O
Kd	O
remained	O
unchanged	O
.	O

However	O
,	O
when	O
PBMC	B-cell_type
were	O
incubated	O
with	O
IL-2	B-protein
alone	O
or	O
IL-4	B-protein
alone	O
,	O
no	O
change	O
in	O
GR-binding	O
affinity	O
was	O
observed	O
.	O

Furthermore	O
,	O
when	O
T	B-cell_type
cells	I-cell_type
and	O
non-T	B-cell_type
cells	I-cell_type
were	O
individually	O
stimulated	O
with	O
combination	O
IL-2	O
+	O
IL-4	O
,	O
a	O
significant	O
reduction	O
in	O
GR-binding	O
affinity	O
was	O
observed	O
only	O
in	O
the	O
T	B-cell_type
cell	I-cell_type
population	I-cell_type
(	O
p	O
=	O
0.0001	O
)	O
.	O

The	O
IL-2	O
+	O
IL-4-induced	O
alteration	O
in	O
PBMC	O
GR	O
Kd	O
was	O
associated	O
with	O
an	O
increase	O
in	O
GR	O
number	O
(	O
8348	O
+/-	O
964	O
vs	O
1710	O
+/-	O
228	O
sites/cell	O
;	O
p	O
=	O
0.0003	O
)	O
.	O

More	O
importantly	O
,	O
the	O
alteration	O
in	O
PBMC	O
GR-binding	O
affinity	O
with	O
IL-2	B-protein
+	O
IL-4	B-protein
was	O
associated	O
with	O
a	O
functional	O
change	O
in	O
T	O
cell	O
response	O
to	O
methylprednisolone	O
MPN	O
,	O
i.e.	O
,	O
a	O
reduced	O
inhibitory	O
effect	O
of	O
MPN	O
on	O
PMA/ionomycin-induced	O
T	B-cell_type
cell	I-cell_type
proliferation	O
.	O

These	O
effects	O
of	O
IL-2	B-protein
+	O
IL-4	B-protein
on	O
PBMC	B-cell_type
GR	B-protein
affinity	O
and	O
response	O
to	O
MPN	O
were	O
blocked	O
by	O
co-incubation	O
with	O
IFN-gamma	B-protein
.	O

Freshly	O
isolated	O
PBMC	B-cell_type
from	O
four	O
patients	O
with	O
SR	O
asthma	O
had	O
a	O
significantly	O
reduced	O
GR-binding	O
affinity	O
(	O
Kd	O
=	O
40.0	O
+/-	O
2.68	O
nM	O
;	O
p	O
=	O
0.0001	O
)	O
when	O
compared	O
with	O
seven	O
normal	O
subjects	O
(	O
7.15	O
+/-	O
0.41	O
nM	O
)	O
.	O

The	O
altered	O
PBMC	O
GR	O
binding	O
from	O
patients	O
with	O
SR	O
asthma	O
reversed	O
to	O
normal	O
when	O
incubated	O
with	O
medium	O
alone	O
,	O
but	O
was	O
sustained	O
with	O
IL-2	B-protein
+	O
IL-4	B-protein
.	O

These	O
observations	O
suggest	O
that	O
with	O
persistent	O
inflammation	O
certain	O
cytokines	B-protein
may	O
contribute	O
to	O
an	O
impaired	O
response	O
to	O
glucocorticoids	O
.	O

Furthermore	O
,	O
the	O
effects	O
of	O
IL-2	B-protein
and	O
IL-4	B-protein
were	O
blocked	O
by	O
IFN-gamma	B-protein
.	O

-DOCSTART-	O

Characterization	O
of	O
the	O
human	B-DNA
CD4	I-DNA
gene	I-DNA
promoter	I-DNA
:	O
transcription	O
from	O
the	O
CD4	B-DNA
gene	I-DNA
core	I-DNA
promoter	I-DNA
is	O
tissue-specific	O
and	O
is	O
activated	O
by	O
Ets	B-protein
proteins	I-protein
.	O

We	O
analyzed	O
the	O
5	B-DNA
'	I-DNA
transcription	I-DNA
control	I-DNA
sequences	I-DNA
of	O
the	O
human	B-DNA
CD4	I-DNA
gene	I-DNA
.	O

We	O
located	O
the	O
transcription	B-DNA
initiation	I-DNA
site	I-DNA
and	O
showed	O
that	O
the	O
CD4	B-DNA
core	I-DNA
promoter	I-DNA
(	O
positions	B-DNA
-40	I-DNA
to	I-DNA
+16	I-DNA
)	O
lacks	O
a	O
classical	O
``	O
TATA	O
''	O
or	O
initiator	B-DNA
positioning	I-DNA
consensus	I-DNA
sequence	I-DNA
but	O
directs	O
precise	O
and	O
efficient	O
transcription	O
when	O
coupled	O
to	O
the	O
ubiquitously	O
active	O
simian	B-DNA
virus	I-DNA
40	I-DNA
enhancer	I-DNA
.	O

The	O
transcriptional	O
activity	O
of	O
the	O
CD4	B-DNA
gene	I-DNA
promoter	I-DNA
correlated	O
with	O
CD4	O
expression	O
in	O
various	O
cell	O
types	O
.	O

Interestingly	O
,	O
the	O
CD4	B-DNA
core	I-DNA
promoter	I-DNA
also	O
displayed	O
a	O
tissue-specific	O
transcriptional	O
activity	O
.	O

Within	O
this	O
fragment	O
,	O
three	O
nucleic	O
acid	O
sequences	O
are	O
completely	O
conserved	O
in	O
the	O
murine	B-DNA
CD4	I-DNA
gene	I-DNA
.	O

One	O
of	O
these	O
sequences	O
contains	O
a	O
perfect	B-DNA
ETS	I-DNA
consensus	I-DNA
sequence	I-DNA
.	O

Another	O
ETS	B-DNA
consensus	I-DNA
sequence	I-DNA
is	O
located	O
1060	O
nt	O
upstream	O
.	O

Electrophoretic-mobility-shift	O
assays	O
showed	O
that	O
the	O
core	O
promoter	O
ETS	O
motif	O
binds	O
an	O
Ets-related	B-protein
protein	I-protein
specifically	O
expressed	O
at	O
high	O
levels	O
in	O
CD4+	B-cell_type
cells	I-cell_type
.	O

Moreover	O
,	O
in	O
CD4-	B-cell_type
cells	I-cell_type
,	O
overexpression	O
of	O
Ets-1	B-protein
or	O
Ets-2	B-protein
efficiently	O
and	O
specifically	O
activated	O
transcription	O
from	O
the	O
CD4	B-DNA
promoter	I-DNA
and	O
core	B-DNA
promoter	I-DNA
.	O

These	O
data	O
indicate	O
that	O
Ets	B-protein
transcription	I-protein
factors	I-protein
play	O
a	O
central	O
role	O
in	O
controlling	O
CD4	O
gene	O
expression	O
,	O
by	O
binding	O
to	O
both	O
a	O
classical	B-DNA
remote	I-DNA
site	I-DNA
and	O
an	O
unusual	O
proximal	B-DNA
activator	I-DNA
sequence	I-DNA

-DOCSTART-	O

Glucocorticoid	O
receptor	O
activation	O
and	O
inactivation	O
in	O
c	O
ultured	B-cell_line
human	I-cell_line
lymphocytes	I-cell_line
.	O

Although	O
glucocorticoids	O
are	O
not	O
cytolytic	O
for	O
and	O
do	O
not	O
inhibit	O
the	O
growth	O
of	O
the	O
IM-9	B-cell_line
line	I-cell_line
of	O
cultured	B-cell_line
human	I-cell_line
lymphoblasts	I-cell_line
,	O
these	O
cells	O
have	O
a	O
high	O
steroid-binding	O
capacity	O
.	O

We	O
have	O
used	O
IM-9	B-cell_type
cells	I-cell_type
in	O
order	O
to	O
examine	O
whether	O
unoccupied	B-protein
glucocorticoid	I-protein
receptors	I-protein
are	O
inactivated	O
and	O
activated	O
in	O
intact	B-cell_type
cells	I-cell_type
.	O

when	O
IM-9	B-cell_type
cells	I-cell_type
are	O
incubated	O
in	O
glucose-free	O
medium	O
in	O
a	O
nitrogen	O
atmosphere	O
,	O
both	O
their	O
ability	O
to	O
bind	O
triamcinolone	O
acetonide	O
and	O
their	O
ATP	O
levels	O
decline	O
and	O
,	O
when	O
glucose	O
and	O
oxygen	O
are	O
reintroduced	O
,	O
ATP	O
levels	O
and	O
receptor	O
activity	O
return	O
.	O

The	O
specific	O
glucocorticoid-binding	O
activity	O
of	O
cytosol	O
prepared	O
from	O
cells	O
exposed	O
to	O
various	O
degrees	O
of	O
energy	O
limitation	O
is	O
directly	O
correlated	O
with	O
the	O
ATP	O
content	O
.	O

Receptor	O
activation	O
in	O
intact	B-cell_type
cells	I-cell_type
is	O
rapid	O
and	O
independent	O
of	O
protein	O
synthesis	O
.	O

Cytosol	O
prepared	O
from	O
inactivated	O
cells	O
can	O
not	O
be	O
activated	O
by	O
addition	O
of	O
ATP	O
.	O

The	O
inactivation	O
of	O
glucocorticoid	B-protein
receptors	I-protein
that	O
occurs	O
when	O
cytosol	O
from	O
normal	O
IM-9	B-cell_line
cells	I-cell_line
is	O
incubated	O
at	O
25	O
degrees	O
C	O
is	O
inhibited	O
by	O
molybdate	O
,	O
vanadate	O
,	O
fluoride	O
,	O
ATP	O
,	O
and	O
several	O
other	O
nucleotides	O
.	O

The	O
experiments	O
with	O
intact	B-cell_type
human	I-cell_type
lymphoblasts	I-cell_type
suggest	O
that	O
assays	O
of	O
specific	O
glucocorticoid-binding	O
capacity	O
do	O
not	O
necessarily	O
reflect	O
the	O
cellular	O
content	O
of	O
receptor	B-protein
protein	I-protein
.	O

-DOCSTART-	O

Glucocorticoids	O
and	O
lymphocytes	B-cell_type
.	O

II	O
.	O

Cell	O
cycle-dependent	O
changes	O
in	O
glucocorticoid	B-protein
receptor	I-protein
content	O
.	O

To	O
study	O
variations	O
in	O
glucocorticoid	B-protein
receptor	I-protein
levels	O
during	O
the	O
cell	O
cycle	O
,	O
we	O
have	O
separated	O
mitogen-stimulated	B-cell_line
human	I-cell_line
peripheral	I-cell_line
lymphocytes	I-cell_line
and	O
rat	B-cell_type
lymph	I-cell_type
node	I-cell_type
cells	I-cell_type
by	O
unit	O
gravity	O
sedimentation	O
and	O
measured	O
glucocorticoid	O
binding	O
in	O
the	O
resultant	O
fractions	O
.	O

By	O
morphologic	O
criteria	O
and	O
thymidine	O
incorporation	O
,	O
the	O
fractions	O
were	O
separated	O
into	O
populations	O
of	O
G0	B-cell_type
and	I-cell_type
G1	I-cell_type
phase	I-cell_type
and	I-cell_type
S	I-cell_type
and	I-cell_type
post-S	I-cell_type
phase	I-cell_type
cells	I-cell_type
.	O

A	O
2-	O
to	O
3-fold	O
increase	O
in	O
glucocorticoid	O
receptor	O
sites	O
per	O
cell	O
,	O
for	O
cells	O
in	O
the	O
S	O
and	O
post-S	O
phase	O
over	O
those	O
in	O
G0	O
and	O
G1	O
,	O
was	O
observed	O
with	O
both	O
nonstimulated	B-cell_type
rat	I-cell_type
lymph	I-cell_type
node	I-cell_type
cell	I-cell_type
suspensions	I-cell_type
and	O
concanavalin	B-cell_line
A-stimulated	I-cell_line
human	I-cell_line
peripheral	I-cell_line
lymphocytes	I-cell_line
.	O

These	O
observations	O
together	O
with	O
those	O
from	O
other	O
studies	O
indicate	O
that	O
formation	O
of	O
new	O
glucocorticoid	B-protein
receptors	I-protein
near	O
the	O
S	O
phase	O
may	O
be	O
a	O
general	O
phenomenon	O
in	O
proliferating	B-cell_type
cells	I-cell_type
.	O

We	O
propose	O
that	O
this	O
increase	O
in	O
glucocorticoid	B-protein
receptors	I-protein
during	O
the	O
cell	O
cycle	O
may	O
explain	O
the	O
increase	O
in	O
glucocorticoid	B-protein
receptors	I-protein
in	O
mitogen-stimulated	B-cell_type
lymphocytes	I-cell_type
.	O

-DOCSTART-	O

The	O
leukocyte	B-cell_type
migration	O
inhibition	O
response	O
to	O
certain	O
breast	B-protein
cancer-related	I-protein
antigens	I-protein
(	O
MCF-7	B-protein
and	O
MuMTV	B-protein
)	O
:	O
their	O
potential	O
as	O
discriminants	O
.	O

Certain	O
oncogenic	O
viruses	O
have	O
been	O
implicated	O
in	O
human	O
breast	O
cancer	O
,	O
including	O
the	O
murine	O
mammary	O
tumor	O
virus	O
(	O
MuMTV	O
)	O
and	O
the	O
Mason-Pfizer	O
monkey	O
virus	O
(	O
MPMV	O
)	O
.	O

We	O
have	O
used	O
the	O
leukocyte	O
migration	O
inhibition	O
(	O
LMI	O
)	O
response	O
to	O
assay	O
the	O
response	O
to	O
several	O
potential	O
breast	O
cancer-related	O
antigens	O
,	O
including	O
MuMTV	B-protein
,	O
MPMV	B-protein
,	O
and	O
a	O
breast	B-cell_line
cancer	I-cell_line
cultured	I-cell_line
cell	I-cell_line
line	I-cell_line
,	O
MCF-7	B-cell_line
,	O
in	O
96	O
breast	O
cancer	O
patients	O
,	O
in	O
32	O
women	O
with	O
benign	O
breast	O
disease	O
,	O
and	O
in	O
67	O
normal	O
women	O
.	O

The	O
lowest	O
tenth	O
percentile	O
of	O
control	O
(	O
LMI	O
)	O
responses	O
was	O
used	O
as	O
the	O
cutoff	O
point	O
to	O
designate	O
responders	O
.	O

Breast	O
cancer	O
patients	O
showed	O
significant	O
responses	O
to	O
MuMTV	B-protein
(	O
49	O
%	O
and	O
to	O
MCF-7	O
(	O
50	O
%	O
)	O
,	O
but	O
not	O
to	O
MPMV	B-protein
(	O
29	O
%	O
)	O
.	O

In	O
a	O
paired-antigen	O
study	O
using	O
MuMTV	B-protein
and	O
MCF-7	B-protein
,	O
75	O
%	O
of	O
the	O
breast	O
cancer	O
patients	O
responded	O
,	O
versus	O
18	O
%	O
of	O
the	O
normal	O
women	O
(	O
P	O
less	O
than	O
0.0050	O
)	O
.	O

The	O
potential	O
for	O
this	O
assay	O
to	O
distinguish	O
``	O
normal	O
''	O
from	O
``	O
breast	O
cancer	O
''	O
was	O
analyzed	O
using	O
a	O
migration	O
index	O
derived	O
from	O
discriminant	O
analysis	O
.	O

The	O
ability	O
of	O
the	O
assay	O
to	O
discriminate	O
``	O
normal	O
''	O
from	O
``	O
cancer	O
''	O
was	O
significant	O
(	O
P	O
less	O
than	O
0.001	O
)	O
and	O
showed	O
a	O
sensitivity	O
of	O
detecting	O
``	O
cancer	O
''	O
of	O
75	O
%	O
.	O

The	O
overall	O
responses	O
to	O
MuMTV	B-protein
and	O
MCF-7	B-protein
were	O
analyzed	O
with	O
reference	O
to	O
certain	O
prognostic	O
factors	O
,	O
but	O
showed	O
no	O
relation	O
to	O
age	O
,	O
menstrual	O
status	O
,	O
estrogen	O
receptor	O
status	O
,	O
or	O
stage	O
of	O
disease	O
.	O

The	O
above	O
reactions	O
suggest	O
that	O
a	O
large	O
proportion	O
of	O
breast	O
cancer	O
patients	O
exhibit	O
presensitization	O
to	O
antigenfs	O
found	O
in	O
MuMTV	B-protein
and	O
MCF-7	B-protein
,	O
which	O
may	O
be	O
cross-reactive	O
with	O
antigens	O
in	O
the	O
primary	O
cancer	O
.	O

These	O
responses	O
appear	O
to	O
be	O
independent	O
of	O
major	O
prognostic	O
variables	O
.	O

Further	O
refinement	O
of	O
this	O
assay	O
may	O
yield	O
one	O
which	O
is	O
more	O
highly	O
discriminating	O
for	O
breast	O
cancer	O
.	O

-DOCSTART-	O

Chronic	O
lymphatic	O
leukaemia	O
:	O
cellular	O
effects	O
of	O
glucocorticoids	O
in	O
vitro	O
.	O

Glucocorticoid	B-protein
receptor	I-protein
levels	O
and	O
steroid	O
induced	O
inhibition	O
of	O
nucleic	O
acid	O
precursors	O
have	O
been	O
examined	O
in	O
lymphocytes	B-cell_type
from	O
27	O
patients	O
at	O
different	O
stages	O
of	O
chronic	O
lymphatic	O
leukaemia	O
.	O

No	O
correlation	O
can	O
be	O
found	O
between	O
the	O
level	O
of	O
glucocorticoid	B-protein
receptors	I-protein
and	O
the	O
stage	O
of	O
the	O
disease	O
.	O

On	O
the	O
other	O
hand	O
,	O
a	O
significant	O
difference	O
(	O
P	O
less	O
than	O
0.02	O
)	O
was	O
found	O
between	O
stage	O
O	O
and	O
stage	O
III/IV	O
patients	O
,	O
in	O
terms	O
of	O
the	O
in	O
vitro	O
effect	O
of	O
dexamethasone	O
on	O
[	O
3H	O
]	O
uridine	O
incorporation	O
.	O

-DOCSTART-	O

Pyrrolidine	O
dithiocarbamate	O
inhibits	O
NF-kappa	B-protein
B	I-protein
mobilization	O
and	O
TNF	O
production	O
in	O
human	B-cell_type
monocytes	I-cell_type
.	O

The	O
human	B-DNA
TNF	I-DNA
promoter	I-DNA
contains	O
four	O
potential	O
nuclear	B-DNA
factor-kappa	I-DNA
B	I-DNA
(	I-DNA
NF-kappa	I-DNA
B	I-DNA
)	I-DNA
-binding	I-DNA
sites	I-DNA
,	O
with	O
the	O
strongest	O
binding	O
seen	O
for	O
the	O
-605	O
motif	O
.	O

Nuclear	O
extracts	O
from	O
unstimulated	B-cell_line
cells	I-cell_line
of	O
the	O
human	B-cell_line
monocytic	I-cell_line
cell	I-cell_line
line	I-cell_line
,	O
Mono	B-cell_line
Mac	I-cell_line
6	I-cell_line
,	O
contain	O
one	O
specific	B-protein
binding	I-protein
protein	I-protein
(	O
complex	B-protein
II	I-protein
)	O
,	O
consistent	O
with	O
a	O
constitutive	B-protein
p50	I-protein
homodimer	I-protein
.	O

Stimulation	O
of	O
Mono	B-cell_line
Mac	I-cell_line
6	I-cell_line
cells	I-cell_line
with	O
LPS	O
will	O
increase	O
complex	B-protein
II	I-protein
and	O
will	O
strongly	O
induce	O
a	O
second	O
specific	O
complex	O
(	O
complex	B-protein
I	I-protein
)	O
,	O
which	O
represents	O
the	O
p50/65	B-protein
heterodimer	I-protein
.	O

Treatment	O
of	O
Mono	B-cell_type
Mac	I-cell_type
6	I-cell_type
cells	I-cell_type
with	O
pyrrolidine-dithiocarbamate	O
(	O
PDTC	O
)	O
at	O
300	O
microM	O
will	O
block	O
the	O
LPS-induced	B-protein
complex	I-protein
I	I-protein
almost	O
completely	O
and	O
will	O
reduce	O
complex	B-protein
II	I-protein
to	O
the	O
constitutive	O
level	O
.	O

Binding	O
activity	O
of	O
other	O
nuclear	B-protein
factors	I-protein
that	O
recognize	O
the	O
SP-1	O
and	O
c/EBP	O
motifs	O
of	O
the	O
human	B-DNA
TNF	I-DNA
promoter	I-DNA
is	O
not	O
affected	O
by	O
such	O
treatment	O
.	O

Northern	O
blot	O
analysis	O
demonstrates	O
that	O
PDTC	O
treatment	O
will	O
strongly	O
reduce	O
LPS-induced	B-protein
TNF	I-protein
transcripts	I-protein
.	O

Secreted	O
TNF	B-protein
protein	I-protein
as	O
detected	O
in	O
the	O
Wehi	O
164S/ActD	O
bioassay	O
and	O
in	O
a	O
sandwich	O
immunoassay	O
was	O
similarly	O
reduced	O
by	O
PDTC	O
.	O

Kinetic	O
analyses	O
show	O
that	O
after	O
LPS	O
stimulation	O
,	O
NF-kappa	B-protein
B	I-protein
will	O
peak	O
at	O
1	O
h	O
,	O
TNF	O
transcript	O
prevalence	O
at	O
2	O
h	O
,	O
and	O
TNF	B-protein
protein	I-protein
at	O
4	O
h	O
.	O

PDTC	O
did	O
not	O
shift	O
this	O
response	O
to	O
LPS	O
to	O
a	O
later	O
time	O
,	O
but	O
suppressed	O
NF-kappa	O
B	O
mobilization	O
,	O
TNF	B-protein
transcripts	I-protein
,	O
and	O
TNF	B-protein
protein	I-protein
over	O
the	O
entire	O
8-h	O
observation	O
period	O
.	O

Analysis	O
of	O
freshly	O
isolated	O
,	O
LPS-stimulated	B-cell_type
blood	I-cell_type
monocytes	I-cell_type
showed	O
a	O
similar	O
blockade	O
of	O
NF-kappa	B-protein
B	I-protein
.	O

Furthermore	O
,	O
in	O
these	O
primary	O
cells	O
,	O
induction	O
of	O
TNF	B-protein
transcripts	I-protein
,	O
as	O
determined	O
by	O
Northern	O
blot	O
analysis	O
and	O
by	O
quantitative	O
polymerase	O
chain	O
reaction	O
,	O
was	O
prevented	O
by	O
PDTC	O
as	O
was	O
TNF	B-protein
protein	I-protein
production	O
.	O

These	O
data	O
show	O
that	O
dithiocarbamates	O
can	O
profoundly	O
affect	O
cytokine	O
expression	O
and	O
suggest	O
that	O
NF-kappa	B-protein
B	I-protein
is	O
involved	O
in	O
LPS-induced	O
TNF	O
gene	O
expression	O
in	O
human	B-cell_type
monocytes	I-cell_type
.	O

-DOCSTART-	O

Comparing	O
regions	O
of	O
the	O
Epstein-Barr	B-protein
virus	I-protein
ZEBRA	I-protein
protein	I-protein
which	O
function	O
as	O
transcriptional	B-DNA
activating	I-DNA
sequences	I-DNA
in	O
Saccharomyces	O
cerevisiae	O
and	O
in	O
B	B-cell_type
cells	I-cell_type
.	O

The	O
ZEBRA	B-protein
protein	I-protein
activates	O
expression	O
of	O
Epstein-Barr	O
virus	O
early-lytic-cycle	B-DNA
genes	I-DNA
in	O
human	B-cell_type
B	I-cell_type
lymphocytes	I-cell_type
.	O

Here	O
it	O
is	O
shown	O
that	O
ZEBRA	B-protein
also	O
behaves	O
as	O
a	O
sequence-specific	B-protein
transcriptional	I-protein
activator	I-protein
in	O
Saccharomyces	O
cerevisiae	O
.	O

Deletional	O
mutagenesis	O
defined	O
three	O
regions	O
of	O
ZEBRA	B-protein
that	O
participate	O
in	O
activation	O
in	O
S.	O
cerevisiae	O
.	O

These	O
regions	O
are	O
designated	O
YI	B-protein
(	O
amino	B-protein
acids	I-protein
[	I-protein
aa	I-protein
]	I-protein
1	I-protein
to	I-protein
25	I-protein
)	O
,	O
YII	B-protein
(	O
aa	B-protein
51	I-protein
to	I-protein
102	I-protein
)	O
,	O
and	O
YIII	B-protein
(	O
aa	B-protein
228	I-protein
to	I-protein
245	I-protein
)	O
.	O

Two	O
of	O
the	O
three	O
regions	O
of	O
the	O
native	O
ZEBRA	B-protein
protein	I-protein
act	O
together	O
to	O
mediate	O
activation	O
when	O
assayed	O
on	O
ZEBRA	B-protein
binding	I-protein
sites	I-protein
.	O

However	O
,	O
when	O
fused	O
to	O
the	O
DNA	B-protein
binding	I-protein
domain	I-protein
of	O
GAL	B-protein
4	O
and	O
assayed	O
on	O
GAL4	B-protein
binding	I-protein
sites	I-protein
,	O
regions	B-protein
YII	I-protein
and	I-protein
YIII	I-protein
were	O
each	O
sufficient	O
to	O
confer	O
activation	O
in	O
S.	O
cerevisiae	O
.	O

Regions	O
of	O
ZEBRA	B-protein
which	O
affected	O
activation	O
in	O
S.	O
cerevisiae	O
were	O
also	O
required	O
in	O
human	B-cell_type
B	I-cell_type
lymphocytes	I-cell_type
.	O

The	O
amino-terminal	B-protein
region	I-protein
of	O
ZEBRA	B-protein
(	O
aa	B-protein
1	I-protein
to	I-protein
98	I-protein
)	O
was	O
required	O
for	O
activation	O
both	O
in	O
S.	O
cerevisiae	O
and	O
in	O
human	B-cell_type
B	I-cell_type
cells	I-cell_type
;	O
deletion	O
of	O
the	O
carboxy-terminal	B-protein
18	I-protein
aa	I-protein
also	O
significantly	O
reduced	O
activation	O
in	O
both	O
cell	O
types	O
.	O

Thus	O
,	O
the	O
behavior	O
of	O
ZEBRA	B-protein
in	O
human	B-cell_type
B	I-cell_type
cells	I-cell_type
and	O
S.	O
cerevisiae	O
suggests	O
that	O
the	O
protein	O
contains	O
universal	B-protein
activation	I-protein
motifs	I-protein
that	O
interact	O
with	O
conserved	O
components	O
of	O
the	O
transcription	O
machinery	O
.	O

However	O
,	O
certain	O
deletion	B-protein
mutants	I-protein
of	O
ZEBRA	B-protein
containing	O
mutations	O
in	O
the	O
N-terminal	B-protein
region	I-protein
exhibited	O
discordant	O
behaviors	O
in	O
S.	O
cerevisiae	O
and	O
in	O
B	O
cells	O
.	O

For	O
example	O
,	O
deletion	O
of	O
ZEBRA	B-protein
aa	I-protein
26	I-protein
to	I-protein
51	I-protein
impaired	O
activation	O
to	O
a	O
great	O
extent	O
in	O
B	B-cell_type
cells	I-cell_type
but	O
had	O
little	O
or	O
no	O
effect	O
in	O
S.	O
cerevisiae	O
.	O

The	O
discordant	O
mutants	O
may	O
reflect	O
interactions	O
with	O
a	O
variable	B-protein
domain	I-protein
of	O
a	O
conserved	O
component	O
or	O
unique	O
interactions	O
with	O
specialized	O
components	O
of	O
the	O
basal	O
transcription	O
apparatus	O
in	O
different	O
cells	O
.	O

-DOCSTART-	O

Functional	O
interaction	O
of	O
the	O
v-Rel	B-protein
and	I-protein
c-Rel	I-protein
oncoproteins	I-protein
with	O
the	O
TATA-binding	B-protein
protein	I-protein
and	O
association	O
with	O
transcription	B-protein
factor	I-protein
IIB	I-protein
.	O

Rel	B-protein
family	I-protein
proteins	I-protein
regulate	O
the	O
expression	O
of	O
genes	O
linked	O
to	O
kappa	O
B-binding	O
motifs	O
.	O

Little	O
is	O
known	O
,	O
however	O
,	O
of	O
the	O
mechanism	O
by	O
which	O
they	O
enhance	O
transcription	O
.	O

We	O
have	O
investigated	O
the	O
ability	O
of	O
the	O
v-Rel	B-protein
and	I-protein
c-Rel	I-protein
oncoproteins	I-protein
to	O
interact	O
with	O
components	O
of	O
the	O
basal	O
transcription	O
machinery	O
.	O

Here	O
we	O
report	O
that	O
both	O
the	O
acidic	O
transcription	O
activation	O
domain	O
mapping	O
to	O
the	O
unique	O
C	B-protein
terminus	I-protein
of	O
chicken	B-protein
c-Rel	I-protein
and	O
the	O
F9	B-protein
cell-specific	I-protein
activation	I-protein
region	I-protein
common	O
to	O
both	O
v-Rel	B-protein
and	O
c-Rel	B-protein
interact	O
with	O
the	O
TATA-binding	B-protein
protein	I-protein
(	O
TBP	B-protein
)	O
and	O
transcription	B-protein
factor	I-protein
IIB	I-protein
(	O
TFIIB	B-protein
)	O
in	O
vitro	O
and	O
in	O
vivo	O
.	O

We	O
also	O
demonstrate	O
that	O
TPB	B-protein
interaction	O
with	O
Rel	B-protein
activation	I-protein
regions	I-protein
leads	O
to	O
synergistic	O
activation	O
of	O
transcription	O
of	O
a	O
kappa	B-DNA
B-linked	I-DNA
reporter	I-DNA
gene	I-DNA
.	O

Combined	O
with	O
the	O
observation	O
that	O
the	O
mouse	B-protein
c-Rel	I-protein
and	I-protein
human	I-protein
RelA	I-protein
proteins	I-protein
also	O
interact	O
with	O
TBP	B-protein
and	O
TFIIB	B-protein
in	O
vitro	O
,	O
these	O
results	O
suggest	O
that	O
association	O
with	O
basal	B-protein
transcription	I-protein
factors	I-protein
is	O
important	O
for	O
the	O
transcriptional	O
activities	O
of	O
Rel	B-protein
family	I-protein
proteins	I-protein
.	O

-DOCSTART-	O

p21ras	B-protein
and	O
calcineurin	O
synergize	O
to	O
regulate	O
the	O
nuclear	B-protein
factor	I-protein
of	O
activated	O
T	B-cell_type
cells	I-cell_type
.	O

In	O
T	B-cell_type
lymphocytes	I-cell_type
,	O
triggering	O
of	O
the	O
T	B-protein
cell	I-protein
receptor	I-protein
(	O
TCR	B-protein
)	O
induces	O
several	O
signaling	O
cascades	O
which	O
ultimately	O
synergize	O
to	O
induce	O
the	O
activity	O
of	O
the	O
nuclear	B-protein
factor	I-protein
of	O
activated	O
T	B-cell_type
cells	I-cell_type
(	O
NFAT	B-cell_line
)	O
,	O
a	O
DNA	B-protein
binding	I-protein
complex	I-protein
critical	O
to	O
the	O
inducibility	O
and	O
T	O
cell	O
specificity	O
of	O
the	O
T	B-protein
cell	I-protein
growth	I-protein
factor	I-protein
interleukin	B-protein
2	I-protein
.	O

One	O
immediate	O
consequence	O
of	O
T	O
cell	O
activation	O
via	O
the	O
TCR	B-protein
is	O
an	O
increase	O
in	O
cytosolic	O
calcium	O
.	O

Calcium	O
signals	O
are	O
important	O
for	O
NFAT	B-cell_line
induction	O
,	O
and	O
recent	O
studies	O
have	O
identified	O
calcineurin	B-protein
,	O
a	O
calcium-calmodulin	B-protein
dependent	I-protein
serine-threonine	I-protein
phosphatase	I-protein
,	O
as	O
a	O
prominent	O
component	O
of	O
the	O
calcium	O
signaling	O
pathway	O
in	O
T	B-cell_type
cells	I-cell_type
.	O

A	O
second	O
important	O
molecule	O
in	O
TCR	O
signal	O
transduction	O
is	O
the	O
guanine	B-protein
nucleotide	I-protein
binding	I-protein
protein	I-protein
,	O
p21ras	B-protein
,	O
which	O
is	O
coupled	O
to	O
the	O
TCR	B-protein
by	O
a	O
protein	B-protein
tyrosine	I-protein
kinase	I-protein
dependent	O
mechanism	O
.	O

The	O
experiments	O
presented	O
here	O
show	O
that	O
expression	O
by	O
transfection	O
of	O
mutationally	O
activated	O
calcineurin	B-protein
or	O
activated	O
p21ras	B-protein
alone	O
is	O
insufficient	O
for	O
NFAT	B-cell_type
transactivation	O
.	O

However	O
,	O
coexpression	O
of	O
the	O
activated	O
calcineurin	O
with	O
activated	O
p21ras	O
could	O
mimic	O
TCR	O
signals	O
in	O
NFAT	B-cell_type
induction	O
.	O

These	O
data	O
identify	O
calcineurin	B-protein
and	O
p21ras	B-protein
as	O
cooperative	O
partners	O
in	O
T	B-cell_type
cell	I-cell_type
activation	O
.	O

-DOCSTART-	O

Expression	O
of	O
the	O
chicken	B-protein
GATA	I-protein
factor	I-protein
family	I-protein
during	O
early	O
erythroid	O
development	O
and	O
differentiation	O
.	O

The	O
DNA	O
motif	O
WGATAR	O
has	O
been	O
identified	O
within	O
transcriptional	B-DNA
regulatory	I-DNA
domains	I-DNA
of	O
globin	B-DNA
and	O
other	O
erythroid-specific	B-DNA
genes	I-DNA
and	O
the	O
activator	B-protein
proteins	I-protein
that	O
bind	O
to	O
this	O
regulatory	B-DNA
element	I-DNA
,	O
the	O
GATA	B-protein
factors	I-protein
,	O
belong	O
to	O
a	O
multi-gene	B-protein
family	I-protein
that	O
is	O
expressed	O
in	O
chicken	B-cell_type
erythroid	I-cell_type
cells	I-cell_type
.	O

Here	O
we	O
show	O
that	O
,	O
as	O
in	O
chickens	O
,	O
multiple	O
members	O
of	O
the	O
GATA	B-protein
factor	I-protein
family	I-protein
are	O
expressed	O
in	O
human	B-cell_type
and	I-cell_type
murine	I-cell_type
erythroid	I-cell_type
cells	I-cell_type
.	O

During	O
the	O
early	O
stages	O
of	O
chicken	O
embryogenesis	O
(	O
well	O
before	O
blood	O
island	O
formation	O
)	O
,	O
each	O
of	O
the	O
GATA	B-protein
family	I-protein
members	I-protein
is	O
transcribed	O
with	O
a	O
unique	O
temporal	O
and	O
spatial	O
pattern	O
.	O

In	O
the	O
primitive	B-cell_type
erythroid	I-cell_type
lineage	I-cell_type
,	O
transcription	O
of	O
the	O
embryonic	B-DNA
epsilon-globin	I-DNA
gene	I-DNA
parallels	O
GATA-1	O
expression	O
while	O
the	O
switch	O
to	O
beta-globin	O
transcription	O
in	O
definitive	B-cell_type
erythroid	I-cell_type
cells	I-cell_type
is	O
directly	O
preceded	O
by	O
a	O
pronounced	O
increase	O
in	O
GATA-3	B-protein
accumulation	O
.	O

The	O
timing	O
and	O
pattern	O
of	O
expression	O
of	O
these	O
different	O
mRNAs	B-RNA
during	O
avian	O
erythroid	O
development	O
and	O
differentiation	O
suggests	O
that	O
temporally	O
regulated	O
changes	O
in	O
GATA	B-protein
factor	I-protein
expression	O
are	O
required	O
for	O
vertebrate	O
hematopoiesis	O
.	O

-DOCSTART-	O

Molecular	O
regulation	O
of	O
human	B-protein
interleukin	I-protein
2	I-protein
and	O
T-cell	B-cell_type
function	O
by	O
interleukin	B-protein
4	I-protein
.	O

Distinct	O
functional	O
T-cell	B-cell_type
subsets	I-cell_type
,	O
differing	O
in	O
the	O
patterns	O
of	O
lymphokines	B-protein
produced	O
,	O
regulate	O
cell-mediated	O
and	O
humoral	O
immune	O
responses	O
.	O

The	O
two	O
major	O
types	O
and	O
their	O
principal	O
products	O
,	O
interleukin	B-protein
4	I-protein
and	O
interferon	B-protein
gamma	I-protein
(	O
IL-4	B-protein
and	O
IFN-gamma	B-protein
)	O
,	O
are	O
reciprocally	O
negatively	O
interactive	O
.	O

To	O
analyze	O
the	O
molecular	O
mechanism	O
of	O
IL-4-mediated	O
suppression	O
of	O
cell-mediated	O
immunity	O
we	O
studied	O
its	O
effects	O
on	O
expression	O
of	O
interleukin	B-protein
2	I-protein
(	O
IL-2	B-protein
)	O
and	O
IFN-gamma	B-protein
.	O

IL-4	B-protein
pretreatment	O
of	O
Jurkat	B-cell_type
cells	I-cell_type
prior	O
to	O
stimulation	O
resulted	O
in	O
a	O
decrease	O
in	O
transcription	O
of	O
the	O
IL2	B-DNA
gene	I-DNA
.	O

IL-4	B-protein
suppressed	O
IL-2	O
and	O
IFN-gamma	O
mRNA	O
levels	O
in	O
primary	B-cell_type
human	I-cell_type
T	I-cell_type
cells	I-cell_type
,	O
and	O
addition	O
of	O
anti-CD28	B-protein
antibodies	I-protein
relieved	O
this	O
suppression	O
.	O

Using	O
enhancer-reporter	B-DNA
constructs	I-DNA
,	O
IL-4	B-protein
specifically	O
down-regulated	O
the	O
NFIL-2B	B-DNA
element	I-DNA
.	O

Electrophoretic	O
mobility	O
shift	O
assays	O
using	O
a	O
DNA	O
oligomer	O
containing	O
the	O
NFIL-2B	B-DNA
binding	I-DNA
site	I-DNA
indicated	O
that	O
IL-4	B-protein
inhibited	O
the	O
NFIL-2B	B-protein
complex	I-protein
and	O
that	O
the	O
NFIL-2B	B-protein
DNA	I-protein
binding	I-protein
factor	I-protein
is	O
distinct	O
from	O
AP-	B-protein
1	O
.	O

These	O
results	O
suggest	O
that	O
IL-4	B-protein
may	O
regulate	O
development	O
and	O
function	O
of	O
T-cell	B-cell_type
subsets	I-cell_type
involved	O
in	O
cell-mediated	O
immunity	O
in	O
part	O
by	O
inhibiting	B-protein
factors	I-protein
required	O
for	O
transcription	O
of	O
the	O
IL2	B-DNA
gene	I-DNA

-DOCSTART-	O

Immunochemical	O
differences	O
between	O
glucocorticoid	B-protein
receptors	I-protein
from	O
corticoid-sensitive	B-cell_type
and	I-cell_type
-resistant	I-cell_type
malignant	I-cell_type
lymphocytes	I-cell_type
.	O

We	O
have	O
explored	O
the	O
possibility	O
of	O
using	O
antibodies	B-protein
against	O
purified	O
rat	O
liver	O
glucocorticoid	B-protein
receptors	I-protein
to	O
study	O
the	O
immunochemical	O
properties	O
of	O
glucocorticoid	B-protein
receptors	I-protein
from	O
murine	B-cell_type
and	I-cell_type
human	I-cell_type
malignant	I-cell_type
lymphocytes	I-cell_type
.	O

For	O
this	O
purpose	O
,	O
purified	B-protein
immune	I-protein
immunoglobulin	I-protein
G	I-protein
was	O
covalently	O
linked	O
to	O
Sepharose	B-protein
CL-4B	I-protein
.	O

We	O
then	O
examined	O
the	O
ability	O
of	O
the	O
affinity	O
gel	O
to	O
recognize	O
cytosolic	B-protein
[	I-protein
3H	I-protein
]	I-protein
triamcinolone	I-protein
acetonide-receptor	I-protein
complexes	I-protein
from	O
the	O
corticoid-sensitive	B-cell_line
(	I-cell_line
CS	I-cell_line
)	I-cell_line
and	I-cell_line
-resistant	I-cell_line
strains	I-cell_line
of	I-cell_line
mouse	I-cell_line
lymphoma	I-cell_line
P1798	I-cell_line
,	O
from	O
CS	B-cell_type
lymphocytes	I-cell_type
of	O
patients	O
with	O
chronic	O
lymphatic	O
leukemia	O
,	O
and	O
from	O
a	O
CS	B-cell_line
clone	I-cell_line
of	O
human	B-cell_type
leukemic	I-cell_type
lymphoblasts	I-cell_type
in	O
tissue	O
culture	O
(	O
CH6	B-cell_line
)	O
.	O

Mouse	O
thymus	O
was	O
used	O
as	O
a	O
source	O
of	O
glucocorticoid	B-protein
receptor	I-protein
from	O
normal	B-cell_type
CS	I-cell_type
lymphocytes	I-cell_type
.	O

Whereas	O
the	O
immunoaffinity	O
column	O
retained	O
70	O
to	O
84	O
%	O
of	O
the	O
58-	O
to	O
62-A	O
(	O
Stokes	O
radius	O
)	O
[	B-protein
3H	I-protein
]	I-protein
triamcinolone	I-protein
acetonide-receptor	I-protein
complexes	I-protein
characteristic	O
of	O
the	O
CS	B-cell_line
mouse	I-cell_line
and	I-cell_line
human	I-cell_line
lymphocytes	I-cell_line
,	O
it	O
failed	O
to	O
recognize	O
the	O
27-	O
to	O
28-A	O
(	O
Stokes	O
radius	O
)	O
glucocorticoid	B-protein
receptor	I-protein
present	O
in	O
corticoid-resistant	O
mouse	B-cell_line
lymphoma	I-cell_line
P1798	I-cell_line
cells	I-cell_line
.	O

Therefore	O
,	O
under	O
appropriate	O
experimental	O
conditions	O
,	O
it	O
was	O
possible	O
to	O
demonstrate	O
cross-reactivity	O
between	O
the	O
antiserum	O
against	O
rat	B-protein
liver	I-protein
glucocorticoid	I-protein
receptor	I-protein
and	O
the	O
58-	O
to	O
62-A	O
(	O
Stokes	O
radius	O
)	O
glucocorticoid	B-protein
receptor	I-protein
from	O
species	O
as	O
diverse	O
as	O
mouse	O
and	O
humans	O
.	O

-DOCSTART-	O

Heterogeneity	O
of	O
the	O
in	O
vitro	O
responses	O
to	O
glucocorticoids	O
in	O
acute	O
leukemia	O
.	O

In	O
leukocyte	B-cell_type
population	I-cell_type
freshly	O
isolated	O
from	O
the	O
blood	O
of	O
26	O
patients	O
with	O
acute	O
leukemia	O
,	O
we	O
have	O
measured	O
several	O
parameters	O
including	O
glucocorticoid	B-protein
receptors	I-protein
,	O
nucleoside	O
incorporation	O
,	O
percentage	O
of	O
cells	O
in	O
S	O
phase	O
,	O
and	O
steroid-induced	O
cell	O
lysis	O
.	O

In	O
addition	O
,	O
in	O
some	O
cases	O
,	O
the	O
short-term	O
response	O
to	O
steroid	O
therapy	O
was	O
determined	O
.	O

Although	O
,	O
in	O
all	O
the	O
patients	O
studied	O
,	O
leukocytes	B-cell_type
were	O
found	O
to	O
contain	O
glucocorticoid	B-protein
receptors	I-protein
,	O
we	O
failed	O
to	O
demonstrate	O
any	O
correlation	O
between	O
the	O
level	O
of	O
binding	O
sites	O
and	O
the	O
in	O
vitro	O
or	O
in	O
vivo	O
response	O
to	O
glucocorticoids	O
.	O

This	O
absence	O
of	O
correlation	O
could	O
be	O
in	O
part	O
explained	O
by	O
the	O
marked	O
heterogeneity	O
of	O
the	O
steroid	O
response	O
demonstrated	O
in	O
leukocyte	O
subpopulations	O
.	O

It	O
appears	O
,	O
however	O
,	O
that	O
the	O
degree	O
of	O
steroid	O
action	O
in	O
vitro	O
as	O
well	O
as	O
the	O
extent	O
of	O
spontaneous	O
and	O
dexamethasone-induced	O
cell	O
death	O
may	O
be	O
related	O
to	O
the	O
number	O
of	O
cells	O
in	O
the	O
S	O
phase	O
of	O
the	O
cell	O
cycle	O
.	O

-DOCSTART-	O

Glucocorticoid	B-protein
receptors	I-protein
in	O
cytosol	O
and	O
nuclear	O
extract	O
of	O
human	B-cell_type
leukocytes	I-cell_type
.	O

Cortisol	O
binding	O
by	O
cytosol	O
and	O
0.4	O
M	O
KCl	O
extract	O
of	O
the	O
nuclear	O
fraction	O
of	O
human	B-cell_type
leukocytes	I-cell_type
were	O
studied	O
by	O
gel	O
chromatography	O
and	O
ion	O
exchange	O
filtration	O
on	O
DEAE	O
cellulose	O
.	O

The	O
cytoplasmic	B-protein
cortisol	I-protein
binding	I-protein
protein	I-protein
has	O
a	O
molecular	O
weight	O
95	O
000	O
and	O
the	O
soluble	B-protein
nuclear	I-protein
binding	I-protein
protein	I-protein
50	O
000	O
.	O

The	O
absence	O
of	O
the	O
uptake	O
of	O
radioactive	O
cortisol	O
by	O
isolated	O
nuclei	O
and	O
the	O
apparent	O
requirement	O
of	O
the	O
cytosol	O
for	O
glucocorticoid	O
specific	O
binding	O
in	O
nuclear	O
receptor	O
sites	O
was	O
observed	O
.	O

The	O
association	O
constant	O
characterising	O
the	O
binding	O
of	O
cortisol	O
to	O
cytosol	O
was	O
KA	O
=	O
3.5	O
.	O
10	O
(	O
9	O
)	O
l/mol	O
.	O

-DOCSTART-	O

Corticosteroid-induced	O
lymphopenia	O
,	O
immunosuppression	O
,	O
and	O
body	O
defense	O
.	O

The	O
apparent	O
paradox	O
of	O
heightened	O
adrenal	O
corticosteroid	O
levels	O
associated	O
with	O
reduction	O
in	O
the	O
competence	O
of	O
the	O
body	O
's	O
defensive	O
apparatus	O
to	O
cope	O
with	O
exposure	O
to	O
new	O
microbial	B-protein
antigens	I-protein
is	O
considered	O
.	O

The	O
question	O
is	O
asked	O
how	O
this	O
lowered	O
defensive	O
capability	O
,	O
which	O
occurs	O
in	O
the	O
face	O
of	O
a	O
threat	O
to	O
body	O
integrity	O
,	O
is	O
consistent	O
with	O
Cannon	O
's	O
principals	O
of	O
the	O
``	O
wisdom	O
of	O
the	O
body.	O
''	O

The	O
suggestion	O
is	O
offered	O
that	O
the	O
immunologic	O
response	O
to	O
self-antigens	B-protein
exposed	O
by	O
disease	O
or	O
trauma	O
may	O
be	O
suppressed	O
by	O
corticosteroid	O
to	O
offset	O
the	O
likelihood	O
of	O
autoimmune	O
attack	O
.	O

-DOCSTART-	O

Protein	B-protein
tyrosine	I-protein
kinase	I-protein
activation	O
is	O
required	O
for	O
lipopolysaccharide	O
induction	O
of	O
cytokines	B-protein
in	O
human	B-cell_type
blood	I-cell_type
monocytes	I-cell_type
.	O

Bacterial	O
LPS	O
induce	O
production	O
of	O
cytokines	B-protein
such	O
as	O
IL-1	B-protein
,	O
IL-6	B-protein
,	O
and	O
TNF	B-protein
in	O
mononuclear	B-cell_type
phagocytes	I-cell_type
,	O
and	O
this	O
represents	O
a	O
central	O
component	O
in	O
the	O
pathogenesis	O
of	O
septic	O
shock	O
syndrome	O
.	O

However	O
,	O
the	O
mechanisms	O
by	O
which	O
LPS	O
activates	O
these	O
cells	O
to	O
express	O
cytokines	B-protein
are	O
not	O
completely	O
characterized	O
.	O

The	O
present	O
study	O
addressed	O
the	O
role	O
of	O
different	O
protein	B-protein
kinases	I-protein
in	O
the	O
LPS	O
induction	O
of	O
cytokines	B-protein
.	O

It	O
is	O
shown	O
that	O
LPS	O
induced	O
a	O
12-	O
to	O
16-fold	O
increase	O
in	O
IL-1	O
beta	O
,	O
IL-6	O
,	O
and	O
TNF-alpha	O
mRNA	O
levels	O
,	O
and	O
this	O
was	O
completely	O
or	O
more	O
than	O
80	O
%	O
blocked	O
by	O
the	O
protein	O
tyrosine	O
kinase	O
specific	O
inhibitors	O
herbimycin	O
A	O
and	O
genistein	O
at	O
the	O
concentrations	O
of	O
1.7	O
and	O
37	O
microM	O
,	O
respectively	O
.	O

Protein	B-protein
kinase	I-protein
C	I-protein
inhibition	O
by	O
staurosporine	O
reduced	O
LPS	O
induction	O
of	O
TNF-alpha	B-protein
,	O
whereas	O
it	O
had	O
no	O
effects	O
on	O
IL-6	B-protein
and	O
IL-1	O
beta	O
.	O

Inhibition	O
of	O
protein	B-protein
kinase	I-protein
A	I-protein
by	O
H89	O
reduced	O
IL-6	B-RNA
mRNA	I-RNA
levels	O
but	O
did	O
not	O
detectably	O
change	O
IL-1	O
beta	O
or	O
TNF-alpha	O
mRNA	O
levels	O
.	O

In	O
contrast	O
,	O
LPS	O
did	O
not	O
increase	O
leukemia	B-RNA
inhibitory	I-RNA
factor	I-RNA
mRNA	I-RNA
,	O
which	O
was	O
constitutively	O
expressed	O
and	O
not	O
significantly	O
reduced	O
by	O
these	O
inhibitors	O
.	O

In	O
addition	O
to	O
cytokine	B-RNA
mRNA	I-RNA
levels	O
,	O
LPS-induced	O
IL-6	O
protein	O
synthesis	O
and	O
IL-6	O
bioactivity	O
were	O
also	O
reduced	O
to	O
baseline	O
levels	O
by	O
the	O
PTK	O
inhibitors	O
herbimycin	O
A	O
and	O
genistein	O
.	O

Both	O
PTK	O
inhibitors	O
also	O
reduced	O
the	O
LPS	O
activation	O
of	O
nuclear	B-protein
factor-kappa	I-protein
B	I-protein
(	O
NF-kappa	B-protein
B	I-protein
)	O
,	O
which	O
is	O
a	O
transcription	B-protein
factor	I-protein
involved	O
in	O
the	O
expression	O
of	O
cytokine	B-RNA
genes	I-RNA
such	O
as	O
IL-6	B-DNA
and	O
TNF-alpha	B-DNA
.	O

The	O
activation	O
of	O
NF-kappa	B-protein
B	I-protein
was	O
also	O
reduced	O
by	O
H89	O
,	O
whereas	O
staurosporine	O
had	O
no	O
effect	O
on	O
this	O
response	O
.	O

In	O
summary	O
,	O
these	O
findings	O
suggest	O
that	O
protein	B-protein
kinase	I-protein
C	I-protein
and	O
protein	B-protein
kinase	I-protein
A	I-protein
appear	O
to	O
have	O
selective	O
effects	O
in	O
the	O
LPS	O
induction	O
of	O
cytokines	B-protein
,	O
whereas	O
PTK	B-protein
is	O
required	O
for	O
LPS	O
induction	O
of	O
a	O
broad	O
spectrum	O
of	O
cytokines	B-protein
and	O
NF-kappa	B-protein
B	I-protein
activation	O
in	O
monocytes	B-cell_type
.	O

-DOCSTART-	O

In	O
vivo	O
control	O
of	O
NF-kappa	B-protein
B	I-protein
activation	O
by	O
I	B-protein
kappa	I-protein
B	I-protein
alpha	I-protein
.	O

The	O
transcription	B-protein
factor	I-protein
NF-kappa	B-protein
B	I-protein
is	O
stored	O
in	O
the	O
cytoplasm	O
in	O
complexes	O
with	O
the	O
inhibitor	B-protein
protein	I-protein
I	I-protein
kappa	I-protein
B	I-protein
alpha	I-protein
.	O

It	O
has	O
been	O
shown	O
in	O
vitro	O
that	O
dissociation	O
of	O
I	B-protein
kappa	I-protein
B	I-protein
alpha	I-protein
from	O
these	O
complexes	O
results	O
in	O
active	B-protein
NF-kappa	I-protein
B	I-protein
.	O

In	O
this	O
report	O
we	O
show	O
that	O
lipopolysaccharide	O
(	O
LPS	O
)	O
-induced	O
activation	O
of	O
B	B-cell_type
or	I-cell_type
pre-B	I-cell_type
cells	I-cell_type
results	O
in	O
loss	O
of	O
I	B-protein
kappa	I-protein
B	I-protein
alpha	I-protein
from	O
NF-kappa	B-protein
B	I-protein
complexes	I-protein
in	O
vivo	O
.	O

Many	O
liberated	O
NF-kappa	B-protein
B	I-protein
dimers	I-protein
reached	O
the	O
nucleus	O
,	O
where	O
increased	O
c-rel	B-protein
,	O
p65	O
and	O
p50	O
were	O
detected	O
by	O
immunoblotting	O
and	O
by	O
DNA	O
binding	O
assays	O
.	O

Some	O
liberated	B-protein
dimers	I-protein
were	O
retained	O
in	O
the	O
cytoplasm	O
,	O
however	O
,	O
through	O
binding	O
to	O
newly	O
synthesized	O
I	B-protein
kappa	I-protein
B	I-protein
alpha	I-protein
,	O
a	O
finding	O
which	O
strongly	O
suggests	O
(	O
i	O
)	O
that	O
the	O
LPS-induced	O
signal	O
causes	O
dissociation	O
of	O
complexes	O
rather	O
than	O
preventing	O
their	O
association	O
and	O
(	O
ii	O
)	O
that	O
dissociation	O
results	O
from	O
modification	O
of	O
I	B-protein
kappa	I-protein
B	I-protein
alpha	I-protein
and	O
not	O
of	O
c-rel	B-protein
or	O
p65	B-protein
.	O

No	O
effect	O
of	O
LPS	O
treatment	O
was	O
detected	O
on	O
p105	B-protein
or	O
p100	B-protein
,	O
which	O
also	O
retain	O
rel	B-protein
family	I-protein
members	I-protein
in	O
the	O
cytoplasm	O
.	O

Quite	O
unexpectedly	O
,	O
we	O
also	O
found	O
that	O
in	O
unstimulated	B-cell_type
cells	I-cell_type
there	O
is	O
a	O
constant	O
ongoing	O
process	O
of	O
degradation	O
and	O
replacement	O
of	O
complexed	B-protein
I	I-protein
kappa	I-protein
B	I-protein
alpha	I-protein
.	O

We	O
propose	O
that	O
this	O
turnover	O
results	O
in	O
the	O
low	O
level	O
of	O
active	B-protein
NF-kappa	I-protein
B	I-protein
presumably	O
necessary	O
even	O
in	O
the	O
unstimulated	O
cell	O
,	O
and	O
that	O
the	O
high	O
rate	O
of	O
synthesis	O
of	O
I	B-protein
kappa	I-protein
B	I-protein
alpha	I-protein
provides	O
the	O
ability	O
to	O
turn	O
off	O
NF-kappa	B-protein
B	I-protein
activity	O
rapidly	O
as	O
soon	O
as	O
the	O
activating	O
signal	O
ceases	O
.	O

-DOCSTART-	O

Identification	O
of	O
a	O
killer	B-DNA
cell-specific	I-DNA
regulatory	I-DNA
element	I-DNA
of	O
the	O
mouse	B-DNA
perforin	I-DNA
gene	I-DNA
:	O
an	O
Ets-binding	O
site-homologous	O
motif	O
that	O
interacts	O
with	O
Ets-related	B-protein
proteins	I-protein
.	O

The	O
gene	O
encoding	O
the	O
cytolytic	B-protein
protein	I-protein
perforin	B-protein
is	O
selectively	O
expressed	O
by	O
activated	B-cell_type
killer	I-cell_type
lymphocytes	I-cell_type
.	O

To	O
understand	O
the	O
mechanisms	O
underlying	O
the	O
cell-type-specific	O
expression	O
of	O
this	O
gene	O
,	O
we	O
have	O
characterized	O
the	O
regulatory	O
functions	O
and	O
the	O
DNA-protein	O
interactions	O
of	O
the	O
5'-flanking	B-DNA
region	I-DNA
of	O
the	O
mouse	B-DNA
perforin	I-DNA
gene	I-DNA
(	O
Pfp	B-DNA
)	O
.	O

A	O
region	O
extending	O
from	O
residues	B-DNA
+62	I-DNA
through	I-DNA
-141	I-DNA
,	O
which	O
possesses	O
the	O
essential	O
promoter	O
activity	O
,	O
and	O
regions	O
further	O
upstream	O
,	O
which	O
are	O
able	O
to	O
either	O
enhance	O
or	O
suppress	O
gene	O
expression	O
,	O
were	O
identified	O
.	O

The	O
region	O
between	O
residues	B-DNA
-411	I-DNA
and	I-DNA
-566	I-DNA
was	O
chosen	O
for	O
further	O
characterization	O
,	O
since	O
it	O
contains	O
an	O
enhancer-like	O
activity	O
.	O

We	O
have	O
identified	O
a	O
32-mer	O
sequence	O
(	O
residues	B-DNA
-491	I-DNA
to	I-DNA
-522	I-DNA
)	O
which	O
appeared	O
to	O
be	O
capable	O
of	O
enhancing	O
gene	O
expression	O
in	O
a	O
killer	O
cell-specific	O
manner	O
.	O

Within	O
this	O
segment	O
,	O
a	O
9-mer	O
motif	O
(	O
5'-ACAGGAAGT-3	O
'	O
,	O
residues	B-DNA
-505	I-DNA
to	I-DNA
-497	I-DNA
;	O
designated	O
NF-P	O
motif	O
)	O
,	O
which	O
is	O
highly	O
homologous	O
to	O
the	O
Ets	B-DNA
proto-oncoprotein-binding	I-DNA
site	I-DNA
,	O
was	O
found	O
to	O
interact	O
with	O
two	O
proteins	O
,	O
NF-P1	B-protein
and	O
NF-P2	B-protein
.	O

NF-P2	B-protein
appears	O
to	O
be	O
induced	O
by	O
reagents	O
known	O
to	O
up-regulate	O
the	O
perforin	O
message	O
level	O
and	O
is	O
present	O
exclusively	O
in	O
killer	B-cell_type
cells	I-cell_type
.	O

Electrophoretic	O
mobility	O
shift	O
assay	O
and	O
UV	O
cross-linking	O
experiments	O
revealed	O
that	O
NF-P1	B-protein
and	O
NF-P2	B-protein
may	O
possess	O
common	O
DNA-binding	B-protein
subunits	I-protein
.	O

However	O
,	O
the	O
larger	O
native	O
molecular	O
mass	O
of	O
NF-P1	B-protein
suggests	O
that	O
NF-P1	B-protein
contains	O
an	O
additional	O
non-DNA-binding	B-protein
subunit	I-protein
(	O
s	O
)	O
.	O

In	O
view	O
of	O
the	O
homology	O
between	O
the	O
NF-P	B-protein
motif	I-protein
and	O
other	O
Ets	B-protein
proto-oncoprotein-binding	I-protein
sites	I-protein
,	O
it	O
is	O
postulated	O
that	O
NF-P1	B-protein
and	O
NF-P2	B-protein
belong	O
to	O
the	O
Ets	B-protein
protein	I-protein
family	I-protein
.	O

Results	O
obtained	O
from	O
the	O
binding	O
competition	O
assay	O
,	O
nevertheless	O
,	O
suggest	O
that	O
NF-P1	B-protein
and	O
NF-P2	B-protein
are	O
related	O
to	O
but	O
distinct	O
from	O
Ets	B-protein
proteins	I-protein
,	O
e.g.	O
,	O
Ets-1	B-protein
,	O
Ets-2	B-protein
,	O
and	O
NF-AT/Elf-1	B-protein
,	O
known	O
to	O
be	O
expressed	O
in	O
T	B-cell_type
cells	I-cell_type
.	O

-DOCSTART-	O

The	O
role	O
of	O
NF-kappa	B-DNA
B1	I-DNA
(	I-DNA
p50/p105	I-DNA
)	I-DNA
gene	I-DNA
expression	O
in	O
activation	O
of	O
human	B-cell_type
blood	I-cell_type
T-lymphocytes	I-cell_type
via	O
CD2	B-protein
and	I-protein
CD28	I-protein
adhesion	I-protein
molecules	I-protein
.	O

Stimulation	O
of	O
primary	B-cell_type
human	I-cell_type
T-lymphocytes	I-cell_type
via	O
CD2	B-protein
and	I-protein
CD28	I-protein
adhesion	I-protein
molecules	I-protein
induces	O
a	O
long-lasting	O
proliferation	O
(	O
>	O
3	O
weeks	O
)	O
.	O

This	O
potent	O
activation	O
does	O
not	O
require	O
accessory	O
cells	O
,	O
such	O
as	O
monocytes	B-cell_type
,	O
but	O
depends	O
on	O
persistent	O
interleukin	B-protein
2	I-protein
(	O
IL-2	B-protein
)	O
secretion	O
and	O
receptivity	O
,	O
which	O
is	O
associated	O
with	O
high	O
and	O
prolonged	O
expression	O
of	O
the	O
inducible	O
CD25/IL-2	B-DNA
receptor	I-DNA
alpha	I-DNA
(	I-DNA
IL-2R	I-DNA
alpha	I-DNA
)	I-DNA
chain	I-DNA
gene	I-DNA
.	O

The	O
transcription	B-protein
factor	I-protein
NF-kappa	I-protein
B	I-protein
participates	O
in	O
the	O
regulation	O
of	O
both	O
IL-2	B-DNA
and	I-DNA
IL-2R	I-DNA
alpha	I-DNA
genes	I-DNA
,	O
as	O
well	O
as	O
multiple	B-DNA
cellular	I-DNA
genes	I-DNA
involved	O
in	O
T-cell	O
proliferation	O
.	O

To	O
evaluate	O
the	O
role	O
of	O
NF-kappa	B-protein
B	I-protein
in	O
human	B-cell_type
peripheral	I-cell_type
blood	I-cell_type
T-lymphocytes	I-cell_type
,	O
we	O
previously	O
analyzed	O
the	O
activation	O
of	O
NF-kappa	B-protein
B-related	I-protein
complexes	I-protein
in	O
response	O
to	O
CD2+CD28	O
costimulation	O
.	O

We	O
demonstrated	O
a	O
long-term	O
induction	O
of	O
p50/p65	B-protein
heterodimer	I-protein
,	O
a	O
putative	B-protein
p65/c-Rel	I-protein
heterodimer	I-protein
,	O
and	O
a	O
constitutive	O
nuclear	O
expression	O
of	O
KBF1/p50	B-protein
homodimers	I-protein
.	I-protein

As	O
the	O
role	O
of	O
p50	B-protein
remains	O
unclear	O
,	O
we	O
focused	O
our	O
present	O
study	O
on	O
NF-kappa	B-protein
B1	I-protein
(	O
p50/p105	B-protein
)	O
gene	O
regulation	O
.	O

Using	O
electrophoretic	O
mobility	O
shift	O
assays	O
and	O
Western	O
and	O
Northern	O
blot	O
analyses	O
,	O
we	O
studied	O
NF-kappa	B-DNA
B1	I-DNA
gene	I-DNA
expression	O
during	O
T-cell	O
stimulation	O
via	O
CD2+CD28	B-protein
.	O

We	O
observed	O
a	O
transient	O
4-	O
to	O
5-fold	O
increase	O
of	O
NF-kappa	O
B1	O
gene	O
expression	O
at	O
both	O
the	O
mRNA	O
and	O
protein	O
levels	O
,	O
lasting	O
for	O
at	O
least	O
24	O
h	O
.	O

p50	O
DNA-binding	O
activity	O
apparently	O
stays	O
highly	O
controlled	O
when	O
p105	O
expression	O
is	O
enhanced	O
by	O
a	O
physiological	O
stimulus	O
of	O
peripheral	B-cell_type
blood	I-cell_type
T-cells	I-cell_type
.	O

Partial	O
inhibition	O
of	O
p50	O
and	O
p105	O
expression	O
by	O
NF-kappa	B-protein
B1	I-protein
antisense	O
oligonucleotides	O
significantly	O
reduced	O
T-cell	O
proliferation	O
and	O
CD25/IL-2R	B-protein
alpha	I-protein
cell	O
surface	O
expression	O
.	O

(	O
ABSTRACT	O
TRUNCATED	O
AT	O
250	O
WORDS	O
)	O

-DOCSTART-	O

Vitamin	B-protein
D	I-protein
receptor	I-protein
quantitation	O
in	O
human	B-cell_type
blood	I-cell_type
mononuclear	I-cell_type
cells	I-cell_type
in	O
health	O
and	O
disease	O
.	O

Vitamin	B-protein
D	I-protein
receptor	I-protein
(	O
VDR	O
)	O
concentration	O
was	O
quantitated	O
in	O
human	B-cell_type
peripheral	I-cell_type
blood	I-cell_type
mononuclear	I-cell_type
cells	I-cell_type
(	O
PBMC	B-cell_type
)	O
from	O
patients	O
with	O
absorptive	O
hypercalciuria	O
(	O
AH	O
)	O
and	O
patients	O
with	O
high	O
1	O
,	O
25	O
(	O
OH	O
)	O
2D3	O
due	O
to	O
acquired	O
or	O
transient	O
disease	O
states	O
and	O
the	O
results	O
compared	O
to	O
those	O
in	O
normal	O
subjects	O
.	O

VDR	O
concentration	O
in	O
resting	B-cell_type
cells	I-cell_type
was	O
not	O
different	O
between	O
the	O
three	O
groups	O
and	O
represented	O
constitutive	O
receptor	O
expression	O
of	O
monocytes	B-cell_type
.	O

Following	O
activation	O
with	O
phytohemagglutinin	B-protein
,	O
patients	B-cell_type
with	O
hypercalcitriolemia	O
demonstrated	O
significantly	O
greater	O
VDR	O
concentrations	O
.	O

Patients	O
with	O
AH	O
demonstrated	O
a	O
normal	O
value	O
for	O
the	O
group	O
,	O
but	O
6	O
patients	O
had	O
significantly	O
greater	O
concentrations	O
of	O
VDR	B-protein
despite	O
normal	O
plasma	O
1	O
,	O
25	O
(	O
OH	O
)	O
2D3	O
in	O
four	O
of	O
the	O
patients	O
.	O

Proliferation	O
,	O
as	O
assessed	O
from	O
[	O
3H	O
]	O
thymidine	O
incorporation	O
was	O
inversely	O
correlated	O
with	O
serum	O
1	O
,	O
25	O
(	O
OH	O
)	O
2D3	O
and	O
was	O
significant	O
(	O
R	O
=	O
-0.299	O
,	O
p	O
=	O
0.048	O
)	O
.	O

Taken	O
together	O
,	O
the	O
results	O
suggest	O
that	O
PBMC	B-cell_type
provide	O
a	O
useful	O
system	O
for	O
studying	O
VDR	B-protein
status	O
in	O
transient	O
or	O
acquired	O
states	O
of	O
hypercalcitriolemia	O
.	O

Furthermore	O
,	O
the	O
studies	O
in	O
patients	O
with	O
absorptive	O
hypercalciuria	O
disclosed	O
it	O
to	O
be	O
a	O
heterogeneous	O
disorder	O
,	O
characterized	O
by	O
both	O
vitamin	O
D-dependent	O
and	O
D-independent	O
forms	O
of	O
receptor	O
up-regulation	O
.	O

-DOCSTART-	O

Reactive	O
oxygen	O
intermediates	O
activate	O
NF-kappa	B-protein
B	I-protein
in	O
a	O
tyrosine	O
kinase-dependent	O
mechanism	O
and	O
in	O
combination	O
with	O
vanadate	O
activate	O
the	O
p56lck	B-protein
and	I-protein
p59fyn	I-protein
tyrosine	I-protein
kinases	I-protein
in	O
human	B-cell_type
lymphocytes	I-cell_type
.	O

We	O
have	O
previously	O
observed	O
that	O
ionizing	O
radiation	O
induces	O
tyrosine	O
phosphorylation	O
in	O
human	B-cell_type
B-lymphocyte	I-cell_type
precursors	I-cell_type
by	O
stimulation	O
of	O
unidentified	O
tyrosine	B-protein
kinases	I-protein
and	O
this	O
phosphorylation	O
is	O
substantially	O
augmented	O
by	O
vanadate	O
.	O

Ionizing	O
radiation	O
generates	O
reactive	O
oxygen	O
intermediates	O
(	O
ROI	O
)	O
.	O

Because	O
H2O2	O
is	O
a	O
potent	O
ROI	O
generator	O
that	O
readily	O
crosses	O
the	O
plasma	O
membrane	O
,	O
we	O
used	O
H2O2	O
to	O
examine	O
the	O
effects	O
of	O
ROI	O
on	O
signal	O
transduction	O
.	O

We	O
now	O
provide	O
evidence	O
that	O
the	O
tyrosine	O
kinase	O
inhibitor	O
herbimycin	O
A	O
and	O
the	O
free	O
radical	O
scavenger	O
N-acetyl-cysteine	O
inhibit	O
both	O
radiation-induced	O
and	O
H2O2-induced	O
activation	O
of	O
NF-kappa	B-protein
B	I-protein
,	O
indicating	O
that	O
activation	O
triggered	O
by	O
ROI	O
is	O
dependent	O
on	O
tyrosine	O
kinase	O
activity	O
.	O

H2O2	O
was	O
found	O
to	O
stimulate	O
Ins-1	B-protein
,	I-protein
4	I-protein
,	I-protein
5-P3	I-protein
production	O
in	O
a	O
tyrosine	O
kinase-dependent	O
manner	O
and	O
to	O
induce	O
calcium	O
signals	O
that	O
were	O
greatly	O
augmented	O
by	O
vanadate	O
.	O

The	O
synergistic	O
induction	O
of	O
tyrosine	O
phosphorylation	O
by	O
H2O2	O
plus	O
vanadate	O
included	O
physiologically	O
relevant	O
proteins	O
such	O
as	O
PLC	B-protein
gamma	I-protein
1	I-protein
.	O

Although	O
treatment	O
of	O
cells	O
with	O
H2O2	O
alone	O
did	O
not	O
affect	O
the	O
activity	O
of	O
src	B-protein
family	I-protein
kinases	I-protein
,	O
treatment	O
with	O
H2O2	O
plus	O
vanadate	O
led	O
to	O
activation	O
of	O
the	O
p56lck	B-protein
and	I-protein
p59fyn	I-protein
tyrosine	I-protein
kinases	I-protein
.	O

The	O
combined	O
inhibition	O
of	O
phosphatases	B-protein
and	O
activation	O
of	O
kinases	B-protein
provides	O
a	O
potent	O
mechanism	O
for	O
the	O
synergistic	O
effects	O
of	O
H2O2	O
plus	O
vanadate	O
.	O

Induction	O
of	O
tyrosine	O
phosphorylation	O
by	O
ROI	O
may	O
thus	O
lead	O
to	O
many	O
of	O
the	O
pleiotropic	O
effects	O
of	O
ROI	O
in	O
lymphoid	B-cell_type
cells	I-cell_type
,	O
including	O
downstream	O
activation	O
of	O
PLC	B-protein
gamma	I-protein
1	I-protein
and	O
NF-kappa	B-protein
B	I-protein

-DOCSTART-	O

Influence	O
of	O
sex	B-protein
hormone	I-protein
binding	I-protein
globulin	I-protein
and	O
serum	B-protein
albumin	I-protein
on	O
the	O
conversion	O
of	O
androstenedione	O
to	O
testosterone	O
by	O
human	B-cell_type
erythrocytes	I-cell_type
.	O

The	O
influence	O
of	O
human	B-protein
serum	I-protein
albumin	I-protein
and	O
sex	B-protein
hormone	I-protein
binding	I-protein
globulin	I-protein
(	O
SHBG	B-protein
)	O
on	O
the	O
enzymic	O
conversion	O
of	O
androstenedione	O
to	O
testosterone	O
in	O
human	B-cell_type
erythrocytes	I-cell_type
was	O
investigated	O
in	O
vitro	O
.	O

Total	O
plasma	O
and	O
albumin	B-protein
delayed	O
the	O
conversion	O
rate	O
of	O
androstenedione	O
,	O
while	O
SHBG	B-protein
increased	O
it	O
markedly	O
.	O

The	O
effect	O
of	O
SHBG	B-protein
was	O
largely	O
abolished	O
by	O
heating	O
to	O
60	O
degrees	O
C	O
for	O
1	O
h	O
and	O
by	O
saturating	O
its	O
binding	O
sites	O
by	O
DHT	O
.	O

The	O
effect	O
of	O
both	O
proteins	O
was	O
found	O
to	O
be	O
related	O
to	O
their	O
concentration	O
.	O

It	O
appears	O
that	O
the	O
binding	O
sites	O
of	O
albumin	B-protein
provide	O
a	O
mechanism	O
for	O
retarding	O
androstenedione	O
uptake	O
by	O
the	O
erythrocytes	B-cell_type
and	O
that	O
the	O
high	O
binding	O
affinity	O
of	O
SHBG	B-protein
for	O
testosterone	O
facilitates	O
the	O
diffusion	O
of	O
this	O
steroid	O
out	O
of	O
the	O
cell	O
and	O
thus	O
,	O
displaces	O
the	O
chemical	O
equilibrium	O
within	O
the	O
cell	O
.	O

-DOCSTART-	O

Glucocorticoids	O
and	O
lymphocytes	B-cell_type
.	O

I	O
.	O

Increased	O
glucocorticoid	B-protein
receptor	I-protein
levels	O
in	O
antigen-stimulated	B-cell_line
lymphocytes	I-cell_line
.	O

Recently	O
a	O
2-	O
to	O
3-fold	O
increase	O
in	O
the	O
number	O
of	O
glucocorticoid	B-protein
receptors	I-protein
in	O
human	B-cell_type
peripheral	I-cell_type
lymphocytes	I-cell_type
has	O
been	O
noted	O
after	O
in	O
vitro	O
mitogen	O
stimulation	O
.	O

Here	O
,	O
we	O
extend	O
these	O
observations	O
to	O
in	O
vivo	O
immunization	O
.	O

After	O
unilateral	O
immunization	O
of	O
adrenalectomized	O
male	O
rats	O
,	O
a	O
50	O
%	O
increase	O
in	O
glucocorticoid	O
receptor	O
sites	O
per	O
cell	O
,	O
determined	O
by	O
binding	O
of	O
dexamethasone	O
,	O
was	O
observed	O
in	O
cell	O
suspensions	O
of	O
homolateral	O
lymph	O
nodes	O
over	O
those	O
from	O
the	O
contralateral	O
nonimmunized	O
side	O
of	O
the	O
same	O
animal	O
.	O

The	O
association	O
constant	O
for	O
dexamethasone	O
was	O
similar	O
in	O
both	O
groups	O
,	O
as	O
was	O
the	O
stereospecificity	O
for	O
various	O
steroids	O
,	O
the	O
time	O
course	O
of	O
cytoplasmic	O
and	O
nuclear	O
association	O
,	O
and	O
cytoplasmic-to-nuclear	O
translocation	O
.	O

Despite	O
a	O
50	O
%	O
increase	O
in	O
the	O
number	O
of	O
glucocorticoid	O
receptor	O
sites	O
per	O
cell	O
,	O
the	O
cells	O
from	O
the	O
homolateral	O
and	O
controlateral	O
lymph	O
nodes	O
were	O
equally	O
sensitive	O
to	O
the	O
inhibitory	O
effects	O
of	O
dexamethasone	O
,	O
as	O
determined	O
by	O
measurements	O
of	O
the	O
incorporation	O
of	O
radiolabeled	O
precursors	O
of	O
protein	O
,	O
RNA	O
,	O
and	O
DNA	O
,	O
or	O
measurements	O
of	O
in	O
vitro	O
cell	O
survival	O
.	O

-DOCSTART-	O

Glucocorticoid	O
receptors	O
and	O
actions	O
in	O
rat	B-cell_type
thymocytes	I-cell_type
and	O
immunologically	O
stimulated	O
human	B-cell_type
peripheral	I-cell_type
lymphocytes	I-cell_type
.	O

After	O
reviewing	O
briefly	O
our	O
earlier	O
studies	O
on	O
glucocorticoid	O
receptors	O
and	O
mechanisms	O
in	O
thymus	B-cell_type
cells	I-cell_type
,	O
we	O
have	O
outlined	O
results	O
from	O
the	O
following	O
two	O
areas	O
of	O
current	O
interest	O
in	O
our	O
laboratories	O
:	O
the	O
``	O
life-cycle	O
''	O
of	O
glucocorticoid	O
receptors	O
and	O
complexes	O
in	O
thymus	B-cell_type
cells	I-cell_type
,	O
and	O
the	O
levels	O
of	O
glucocorticoid	O
receptors	O
and	O
sensitivity	O
in	O
immunologically	B-cell_type
stimulated	I-cell_type
human	I-cell_type
peripheral	I-cell_type
lymphocytes	I-cell_type
.	O

Several	O
of	O
our	O
results	O
on	O
energetics	O
and	O
kinetics	O
of	O
hormone	O
binding	O
to	O
glucocorticoid	B-protein
receptors	I-protein
in	O
rat	B-cell_type
thymus	I-cell_type
cells	I-cell_type
seem	O
to	O
require	O
extension	O
of	O
the	O
simplest	O
model	O
of	O
hormone-receptor	O
transformations	O
in	O
intact	B-cell_type
cells	I-cell_type
.	O

ATP-depletion	O
experiments	O
suggest	O
the	O
existence	O
of	O
a	O
nonbinding	O
form	O
of	O
the	O
receptor	O
;	O
``	O
chase	O
''	O
experiments	O
suggest	O
reaction	O
of	O
hormone	O
directly	O
with	O
nuclear-bound	O
receptor	O
;	O
experiments	O
on	O
depletion	O
and	O
replenishment	O
of	O
cytoplasmic	B-protein
receptor	I-protein
using	O
cortisol	O
and	O
dexamethasone	O
suggest	O
the	O
existence	O
of	O
at	O
least	O
two	O
subpopulations	O
of	O
nuclear-bound	B-protein
hormone-receptor	I-protein
complex	I-protein
.	O

We	O
have	O
found	O
that	O
mitogen	O
or	O
immunologic	O
stimulation	O
of	O
human	B-cell_type
peripheral	I-cell_type
lymphocytes	I-cell_type
in	O
culture	O
leads	O
within	O
24	O
h	O
or	O
so	O
to	O
a	O
striking	O
increase	O
in	O
the	O
number	O
of	O
glucocorticoid	B-protein
receptor	I-protein
sites	O
per	O
cell	O
.	O

We	O
believe	O
this	O
increase	O
may	O
be	O
due	O
to	O
partial	O
synchronization	O
of	O
the	O
cell	O
population	O
in	O
a	O
phase	O
of	O
the	O
cell	O
cycle	O
in	O
which	O
receptor	O
content	O
is	O
high	O
.	O

Contrary	O
to	O
the	O
widely	O
held	O
view	O
that	O
mitogen-stimulated	B-cell_type
cells	I-cell_type
become	O
insensitive	O
to	O
glucocorticoids	O
,	O
our	O
experiments	O
show	O
that	O
with	O
respect	O
to	O
inhibition	O
of	O
thymidine	O
and	O
uridine	O
incorporation	O
and	O
glucose	O
uptake	O
,	O
the	O
cells	O
are	O
highly	O
sensitive	O
to	O
dexamethasone	O
at	O
24	O
,	O
48	O
,	O
and	O
72	O
h	O
after	O
stimulation	O
with	O
concanavalin	B-protein
A	I-protein
.	O

-DOCSTART-	O

Nuclear	O
glucocorticoid	O
binding	O
in	O
chronic	B-cell_type
lymphatic	I-cell_type
leukemia	I-cell_type
lymphocytes	I-cell_type
.	O

A	O
reliable	O
procedure	O
is	O
described	O
for	O
isolating	O
3H-triamcinolone	B-protein
acetonide	I-protein
(	I-protein
3H-TA	I-protein
)	I-protein
receptor	I-protein
complexes	I-protein
from	O
purified	O
chronic	O
lymphatic	B-cell_type
leukemia	I-cell_type
(	I-cell_type
CLL	I-cell_type
)	I-cell_type
lymphocyte	I-cell_type
nuclei	O
,	O
based	O
on	O
the	O
use	O
of	O
carbobenzoxy-L-phenylalanine	O
(	O
CBZ-L-phe	O
)	O
to	O
prevent	O
breakdown	O
of	O
hormone-receptor	B-protein
complexes	I-protein
during	O
extraction	O
of	O
nuclei	O
with	O
0.6M	O
KCl	O
.	O

Using	O
this	O
method	O
,	O
specific	O
nuclear	O
glucocorticoid	O
binding	O
was	O
demonstrated	O
in	O
14/14	O
patients	O
with	O
untreated	O
CLL	O
.	O

No	O
correlation	O
was	O
found	O
between	O
levels	O
of	O
nuclear-associated	O
3H-TA	O
and	O
peripheral	O
white	O
blood	O
cell	O
count	O
or	O
rosetting	O
ability	O
of	O
circulating	B-cell_type
lymphocytes	I-cell_type
.	O

-DOCSTART-	O

Multiple	O
closely-linked	O
NFAT/octamer	B-DNA
and	O
HMG	B-DNA
I	I-DNA
(	I-DNA
Y	I-DNA
)	I-DNA
binding	I-DNA
sites	I-DNA
are	O
part	O
of	O
the	O
interleukin-4	O
promoter	O
.	O

We	O
show	O
here	O
that	O
the	O
immediate	B-DNA
upstream	I-DNA
region	I-DNA
(	O
from	O
position	O
-12	O
to	O
-270	O
)	O
of	O
the	O
murine	B-DNA
interleukin	I-DNA
4	I-DNA
(	I-DNA
Il-4	I-DNA
)	I-DNA
gene	I-DNA
harbors	O
a	O
strong	O
cell-type	O
specific	O
transcriptional	B-DNA
enhancer	I-DNA
.	O

In	O
T	B-cell_type
lymphoma	I-cell_type
cells	I-cell_type
,	O
the	O
activity	O
of	O
the	O
Il-4	B-DNA
promoter/enhancer	I-DNA
is	O
stimulated	O
by	O
phorbol	O
esters	O
,	O
Ca++	O
ionophores	O
and	O
agonists	O
of	O
protein	B-protein
kinase	I-protein
A	I-protein
and	O
inhibited	O
by	O
low	O
doses	O
of	O
the	O
immunosuppressant	O
cyclosporin	O
A	O
.	O

The	O
Il-4	B-DNA
promoter/enhancer	I-DNA
is	O
transcriptionally	O
inactive	O
in	O
B	B-cell_type
lymphoma	I-cell_type
cells	I-cell_type
and	O
HeLa	B-cell_type
cells	I-cell_type
.	O

DNase	O
I	O
footprint	O
protection	O
experiments	O
revealed	O
six	O
sites	O
of	O
the	O
Il-4	B-DNA
promoter/enhancer	I-DNA
to	O
be	O
bound	O
by	O
nuclear	B-protein
proteins	I-protein
from	O
lymphoid	B-cell_type
and	I-cell_type
myeloid	I-cell_type
cells	I-cell_type
.	O

Among	O
them	O
are	O
four	O
purine	O
boxes	O
which	O
have	O
been	O
described	O
to	O
be	O
important	O
sequence	O
motifs	O
of	O
the	O
Il-2	B-DNA
promoter	I-DNA
.	O

They	O
contain	O
the	O
motif	O
GGAAA	O
and	O
are	O
recognized	O
by	O
the	O
inducible	B-protein
and	I-protein
cyclosporin	I-protein
A-sensitive	I-protein
transcription	I-protein
factor	I-protein
NFAT-1	I-protein
.	O

Three	O
of	O
the	O
Il-4	B-DNA
NFAT-1	I-DNA
sites	I-DNA
are	O
closely	O
linked	O
to	O
weak	O
binding	O
sites	O
of	O
Octamer	B-protein
factors	I-protein
.	O

Several	O
purine	O
boxes	O
and	O
an	O
AT-rich	B-DNA
protein-binding	I-DNA
site	I-DNA
of	O
the	O
Il-4	O
promoter	O
are	O
also	O
recognized	O
by	O
the	O
high	B-protein
mobility	I-protein
group	I-protein
protein	I-protein
HMG	B-protein
I	I-protein
(	I-protein
Y	I-protein
)	O
.	O

Whereas	O
the	O
binding	O
of	O
NFAT-1	B-protein
and	O
Octamer	B-protein
factors	I-protein
enhance	O
the	O
activity	O
of	O
the	O
Il-4	B-DNA
promoter	I-DNA
,	O
the	O
binding	O
of	O
HMG	O
I	O
(	O
Y	O
)	O
suppresses	O
its	O
activity	O
and	O
,	O
therefore	O
,	O
appears	O
to	O
be	O
involved	O
in	O
the	O
suppression	O
of	O
Il-4	B-protein
transcription	O
in	O
resting	B-cell_type
T	I-cell_type
lymphocytes	I-cell_type
.	O

-DOCSTART-	O

Reversibility	O
of	O
the	O
differentiated	O
state	O
in	O
somatic	B-cell_type
cells	I-cell_type
.	O

Analysis	O
of	O
de	O
novo	O
gene	O
activation	O
in	O
multinucleated	B-cell_type
heterokaryons	I-cell_type
has	O
shown	O
that	O
the	O
differentiated	O
state	O
,	O
although	O
stable	O
,	O
is	O
not	O
irreversible	O
,	O
and	O
can	O
be	O
reprogrammed	O
in	O
the	O
presence	O
of	O
appropriate	O
combinations	O
of	O
trans-acting	B-protein
regulatory	I-protein
molecules	I-protein
.	O

These	O
properties	O
have	O
been	O
exploited	O
to	O
design	O
strategies	O
for	O
identifying	O
novel	O
regulators	O
of	O
cellular	O
differentiation	O
.	O

-DOCSTART-	O

Phosphatidylcholine	O
hydrolysis	O
activates	O
NF-kappa	B-protein
B	I-protein
and	O
increases	O
human	O
immunodeficiency	O
virus	O
replication	O
in	O
human	B-cell_type
monocytes	I-cell_type
and	O
T	B-cell_type
lymphocytes	I-cell_type
.	O

We	O
have	O
tested	O
whether	O
breakdown	O
of	O
phosphatidylcholine	O
(	O
PC	O
)	O
initiated	O
by	O
exogenous	O
addition	O
of	O
a	O
PC-specific	B-protein
phospholipase	I-protein
C	I-protein
(	O
PC-PLC	B-protein
)	O
from	O
Bacillus	O
cereus	O
or	O
by	O
endogenous	O
overexpression	O
of	O
PC-PLC	B-protein
induces	O
functional	O
activation	O
of	O
NF-kappa	B-protein
B	I-protein
and	O
increases	O
human	B-DNA
immunodeficiency	I-DNA
virus	I-DNA
(	I-DNA
HIV	I-DNA
)	I-DNA
enhancer	I-DNA
activity	O
.	O

PC-PLC-activated	O
hydrolysis	O
of	O
PC	O
was	O
found	O
to	O
induce	O
bona	O
fide	O
p50/p65	B-protein
NF-kappa	I-protein
B	I-protein
binding	O
activity	O
in	O
three	O
different	O
cell	B-cell_line
lines	I-cell_line
of	O
human	O
or	O
murine	O
origin	O
.	O

No	O
significant	O
changes	O
in	O
the	O
turnover	O
of	O
other	O
cellular	O
phospholipids	O
were	O
detected	O
in	O
PC-PLC-treated	B-cell_line
cells	I-cell_line
.	O

Induction	O
of	O
NF-kappa	B-protein
B	I-protein
by	O
PC-PLC	B-protein
did	O
not	O
depend	O
on	O
de	O
novo	O
synthesis	O
of	O
proteins	O
or	O
autocrine	O
secretion	O
of	O
either	O
tumor	B-protein
necrosis	I-protein
factor	I-protein
or	O
interleukin	B-protein
1	I-protein
.	O

In	O
human	B-cell_line
monocytic	I-cell_line
and	I-cell_line
lymphoblastoid	I-cell_line
T-cell	I-cell_line
lines	I-cell_line
,	O
induction	O
of	O
NF-kappa	B-protein
B	I-protein
by	O
PC-PLC	B-protein
resulted	O
in	O
clear	O
induction	O
of	O
luciferase	B-protein
expression	O
vectors	O
placed	O
under	O
the	O
control	O
of	O
synthetic	B-DNA
kappa	I-DNA
B	I-DNA
enhancers	I-DNA
or	O
wild	O
type	O
,	O
but	O
not	O
kappa	O
B-mutated	O
,	O
HIV	B-DNA
long	I-DNA
terminal	I-DNA
repeat	I-DNA
constructs	I-DNA
.	O

HIV	O
replication	O
was	O
increased	O
by	O
PC-PLC	B-protein
in	O
chronically	O
infected	O
monocytes	B-cell_type
and	O
T	O
lymphocytes	B-cell_type
.	O

NF-kappa	O
B	O
activation	O
promoted	O
by	O
addition	O
of	O
exogenous	B-protein
PC-PLC	I-protein
correlated	O
with	O
an	O
intense	O
production	O
of	O
diacylglycerol	O
.	O

However	O
,	O
addition	O
of	O
a	O
phosphatidylinositol-specific	B-protein
PLC	I-protein
from	O
B.cereus	O
also	O
induced	O
diacylglycerol	O
but	O
did	O
not	O
activate	O
kappa	B-protein
B	I-protein
enhancer-directed	I-protein
vectors	I-protein
.	O

PC-PLC-induced	O
NF-kappa	O
B	O
activation	O
could	O
not	O
be	O
blocked	O
by	O
a	O
specific	O
inhibitor	O
of	O
phorbol	B-protein
ester-inducible	I-protein
protein	I-protein
kinases	I-protein
C	I-protein
.	O

These	O
results	O
indicate	O
that	O
a	O
cellular	O
transduction	O
pathway	O
,	O
dependent	O
on	O
specific	O
PC	O
breakdown	O
,	O
is	O
functional	O
in	O
T	B-cell_type
lymphocytes	I-cell_type
and	O
monocytes	B-cell_type
and	O
may	O
be	O
used	O
by	O
various	O
transmembrane	O
receptors	O
to	O
activate	O
HIV	O
transcription	O
through	O
NF-kappa	O
B-dependent	O
induction	O
of	O
the	O
HIV	B-DNA
enhancer	I-DNA
.	O

-DOCSTART-	O

Novel	O
mechanism	O
for	O
inhibition	O
of	O
human	B-cell_type
T	I-cell_type
cells	I-cell_type
by	O
glucocorticoids	O
.	O

Glucocorticoids	O
inhibit	O
signal	O
transduction	O
through	O
IL-2	B-protein
receptor	I-protein
.	O

Interaction	O
of	O
IL-2	B-protein
with	O
its	O
high	B-protein
affinity	I-protein
membrane	I-protein
receptor	I-protein
complex	I-protein
(	O
IL-2R	B-protein
)	O
present	O
on	O
activated	O
T	B-cell_type
lymphocytes	I-cell_type
induces	O
cell	O
proliferation	O
and	O
mediates	O
effector	O
functions	O
.	O

Glucocorticoids	O
inhibit	O
IL-2	B-protein
production	O
by	O
inhibiting	O
TCR-mediated	O
signal	O
transduction	O
.	O

We	O
asked	O
whether	O
they	O
also	O
inhibit	O
the	O
action	O
of	O
IL-2	B-protein
by	O
inhibiting	O
signal	O
transduction	O
through	O
IL-2R	B-protein
.	O

Human	B-cell_type
peripheral	I-cell_type
blood	I-cell_type
T	I-cell_type
cells	I-cell_type
,	O
stimulated	O
with	O
PMA	O
for	O
48	O
h	O
(	O
PMA	B-cell_line
blasts	I-cell_line
)	O
,	O
were	O
incubated	O
with	O
IL-2	B-protein
in	O
the	O
presence	O
of	O
incremental	O
dosages	O
of	O
dexamethasone	O
(	O
Dex	O
;	O
10	O
(	O
-5	O
)	O
-10	O
(	O
-9	O
)	O
M	O
)	O
.	O

Dex	O
inhibited	O
the	O
IL-2-dependent	O
proliferation	O
of	O
PMA	B-cell_line
blasts	I-cell_line
in	O
a	O
dose-dependent	O
fashion	O
(	O
IC50	O
,	O
5	O
x	O
10	O
(	O
-8	O
)	O
M	O
)	O
.	O

Cell	O
surface	O
expression	O
of	O
IL-2R	B-protein
alpha-	I-protein
and	I-protein
beta-chains	I-protein
as	O
determined	O
by	O
immunofluorescence	O
analysis	O
was	O
not	O
affected	O
by	O
Dex	O
.	O

In	O
addition	O
,	O
Scatchard	O
plot	O
analysis	O
of	O
125I-labeled	B-protein
IL-2	I-protein
showed	O
that	O
Dex	O
did	O
not	O
affect	O
the	O
binding	O
of	O
IL-2	B-protein
,	O
thus	O
suggesting	O
that	O
inhibition	O
is	O
due	O
to	O
a	O
postreceptor	O
effect	O
.	O

Inhibition	O
of	O
T	O
cell	O
proliferation	O
by	O
Dex	O
was	O
associated	O
with	O
decreased	O
IL-2-dependent	O
tyrosine	O
phosphorylation	O
of	O
several	O
intracellular	B-protein
proteins	I-protein
and	O
decreased	O
phosphorylation	O
of	O
the	O
retinoblastoma	B-protein
gene	I-protein
product	I-protein
Rb	I-protein
,	O
a	O
protein	O
essential	O
for	O
controlling	O
the	O
progression	O
of	O
cells	O
through	O
the	O
cell	O
cycle	O
.	O

IL-2-dependent	O
IL-2R	O
alpha	O
expression	O
in	O
PMA	B-cell_line
blasts	I-cell_line
and	O
NF-kB	B-protein
induction	O
in	O
resting	B-cell_type
human	I-cell_type
T	I-cell_type
cells	I-cell_type
were	O
also	O
inhibited	O
by	O
Dex	O
.	O

These	O
results	O
demonstrate	O
that	O
glucocorticoids	O
inhibit	O
preactivated	B-cell_line
T	I-cell_line
cells	I-cell_line
by	O
down-regulating	O
signal	O
transduction	O
through	O
IL-2R	B-protein
.	O

-DOCSTART-	O

Chronic	O
human	O
immunodeficiency	O
virus	O
type	O
1	O
infection	O
stimulates	O
distinct	O
NF-kappa	O
B/rel	O
DNA	O
binding	O
activities	O
in	O
myelomonoblastic	B-cell_type
cells	I-cell_type
.	O

The	O
relationship	O
between	O
human	O
immunodeficiency	O
virus	O
type	O
1	O
(	O
HIV-1	O
)	O
infection	O
and	O
the	O
induction	O
of	O
NF-kappa	O
B	O
binding	O
activity	O
was	O
examined	O
in	O
a	O
myeloid	B-cell_line
cell	I-cell_line
model	I-cell_line
of	O
HIV-1	O
infection	O
derived	O
from	O
the	O
PLB-985	B-cell_line
cell	I-cell_line
line	I-cell_line
.	O

Chronic	O
infection	O
of	O
PLB-985	B-cell_line
cells	I-cell_line
led	O
to	O
increased	O
monocyte-specific	O
surface	O
marker	O
expression	O
,	O
increased	O
c-fms	O
gene	O
transcription	O
,	O
and	O
morphological	O
alterations	O
consistent	O
with	O
differentiation	O
along	O
the	O
monocytic	O
pathway	O
.	O

PLB-IIIB	B-cell_line
cells	I-cell_line
displayed	O
a	O
constitutive	O
NF-kappa	O
B-like	O
binding	O
activity	O
that	O
was	O
distinct	O
from	O
that	O
induced	O
by	O
tumor	B-protein
necrosis	I-protein
factor	I-protein
alpha	I-protein
or	O
phorbol	O
12-myristate	O
13-acetate	O
treatment	O
of	O
the	O
parental	B-cell_line
PLB-985	I-cell_line
cell	I-cell_line
line	I-cell_line
.	O

This	O
unique	O
DNA	O
binding	O
activity	O
consisted	O
of	O
proteins	O
of	O
70	B-protein
,	I-protein
90	I-protein
,	I-protein
and	I-protein
100	I-protein
kDa	I-protein
with	O
a	O
high	O
degree	O
of	O
binding	O
specificity	O
for	O
the	O
NF-kappa	B-protein
B	I-protein
site	I-protein
within	O
the	O
PRDII	B-protein
domain	I-protein
of	O
beta	B-protein
interferon	I-protein
.	O

In	O
this	O
report	O
,	O
we	O
characterize	O
the	O
nature	O
of	O
these	O
proteins	O
and	O
demonstrate	O
that	O
binding	O
of	O
these	O
proteins	O
is	O
also	O
induced	O
following	O
Sendai	O
paramyxovirus	O
infection	O
.	O

The	O
70-kDa	B-protein
protein	I-protein
corresponds	O
to	O
the	O
NF-kappa	B-protein
B	I-protein
RelA	I-protein
(	I-protein
p65	I-protein
)	I-protein
subunit	I-protein
,	O
which	O
is	O
activated	O
in	O
response	O
to	O
an	O
acute	O
paramyxovirus	O
infection	O
or	O
a	O
chronic	O
HIV-1	O
infection	O
.	O

Virus	O
infection	O
does	O
not	O
appear	O
to	O
alter	O
the	O
amount	O
of	O
RelA	B-protein
(	O
p65	B-protein
)	O
or	O
NFKB1	B-protein
(	O
p50	B-protein
)	O
but	O
rather	O
affects	O
the	O
capacity	O
of	O
I	B-protein
kappa	I-protein
B	I-protein
alpha	I-protein
to	O
sequester	O
RelA	B-protein
(	I-protein
p65	I-protein
)	I-protein
,	O
therefore	O
leading	O
to	O
constitutive	O
levels	O
of	O
RelA	O
DNA	O
binding	O
activity	O
and	O
to	O
increased	O
levels	O
of	O
NF-kappa	O
B-dependent	O
gene	O
activity	O
.	O

The	O
virally	O
induced	O
90-	B-protein
to	I-protein
100-kDa	I-protein
proteins	I-protein
have	O
a	O
distinct	O
binding	O
specificity	O
for	O
the	O
PRDII	B-DNA
domain	I-DNA
and	O
an	O
AT-rich	B-DNA
sequence	I-DNA
but	O
do	O
not	O
cross-react	O
with	O
NF-kappa	O
B	O
subunit-specific	O
antisera	O
directed	O
against	O
NFKB1	B-protein
(	O
p105	B-protein
or	O
p50	B-protein
)	O
,	O
NFKB2	B-protein
(	O
p100	B-protein
or	O
p52	B-protein
)	O
,	O
RelA	B-protein
(	O
p65	B-protein
)	O
,	O
or	O
c-rel	B-protein
.	O

DNA	O
binding	O
of	O
the	O
90-	B-protein
to	I-protein
100-kDa	I-protein
proteins	I-protein
was	O
not	O
inhibited	O
by	O
recombinant	B-protein
I	I-protein
kappa	I-protein
B	I-protein
alpha/MAD-3	I-protein
and	O
was	O
resistant	O
to	O
tryptic	O
digestion	O
,	O
suggesting	O
that	O
these	O
proteins	O
may	O
not	O
be	O
NF-kappa	B-protein
B	I-protein
related	I-protein
.	O

Transient	O
cotransfection	O
experiments	O
demonstrated	O
that	O
RelA	O
and	O
NFKB1	O
expression	O
maximally	O
stimulated	O
HIV-1	B-DNA
LTR-	I-DNA
and	I-DNA
NF-kappa	I-DNA
B-dependent	I-DNA
reporter	I-DNA
genes	I-DNA
;	O
differences	O
in	O
NF-kappa	O
B-like	O
binding	O
activity	O
were	O
also	O
reflected	O
in	O
higher	O
constitutive	O
levels	O
of	O
NF-kappa	O
B-regulated	O
gene	O
expression	O
in	O
HIV-1-infected	B-cell_line
myeloid	I-cell_line
cells	I-cell_line
.	O

-DOCSTART-	O

Presence	O
of	O
estrogen-binding	O
sites	O
on	O
macrophage-like	B-cell_type
synoviocytes	I-cell_type
and	O
CD8+	B-cell_line
,	I-cell_line
CD29+	I-cell_line
,	I-cell_line
CD45RO+	I-cell_line
T	I-cell_line
lymphocytes	I-cell_line
in	O
normal	O
and	O
rheumatoid	O
synovium	O
.	O

OBJECTIVE	O
.	O

To	O
study	O
the	O
presence	O
of	O
estrogen-binding	O
sites	O
(	O
EBS	O
)	O
in	O
the	O
synovial	O
tissues	O
of	O
male	O
and	O
female	O
patients	O
with	O
rheumatoid	O
arthritis	O
(	O
RA	O
)	O
and	O
in	O
age-	O
and	O
sex-matched	O
healthy	O
controls	O
.	O

METHODS	O
.	O

Both	O
type	O
1	O
(	O
high	O
affinity	O
,	O
low	O
binding	O
capacity	O
)	O
and	O
type	O
2	O
(	O
reduced	O
affinity	O
,	O
higher	O
binding	O
capacity	O
)	O
EBS	O
were	O
investigated	O
in	O
both	O
soluble	O
and	O
nuclear	O
fractions	O
of	O
homogenized	O
synovial	O
tissue	O
samples	O
by	O
a	O
dextran-coated	O
charcoal	O
method	O
.	O

To	O
determine	O
what	O
type	O
of	O
synovial	B-cell_type
cell	I-cell_type
was	O
positive	O
for	O
EBS	O
,	O
cryosections	O
of	O
synovial	O
tissues	O
were	O
immunostained	O
with	O
a	O
specific	O
monoclonal	B-protein
anti-estrogen	I-protein
receptor	I-protein
antibody	I-protein
(	O
anti-ER	B-protein
MAb	I-protein
)	O
using	O
both	O
immunofluorescence	O
and	O
immunoperoxidase	O
techniques	O
.	O

Double	O
immunostaining	O
with	O
the	O
anti-ER	B-protein
MAb	I-protein
and	O
with	O
specific	B-protein
MAb	I-protein
to	O
detect	O
different	O
macrophage	B-protein
antigens	I-protein
(	O
Ber-MAC3	B-protein
,	O
MAC387	B-protein
,	O
CD68	B-protein
)	O
and	O
CD8+	B-cell_line
T	I-cell_line
cell	I-cell_line
subsets	I-cell_line
(	O
CD29+	B-cell_line
,	O
CD45RO+	B-cell_line
and	O
CD29-	B-cell_line
,	O
CD45RO-	B-cell_line
)	O
was	O
performed	O
.	O

RESULTS	O
.	O

Higher	O
affinity	O
EBS	O
were	O
found	O
mostly	O
in	O
nuclear	O
cell	O
fractions	O
of	O
either	O
RA	O
or	O
control	O
synovial	O
tissues	O
(	O
28	O
of	O
the	O
33	O
)	O
.	O

These	O
EBS	O
were	O
present	O
to	O
a	O
lesser	O
extent	O
in	O
soluble	O
cell	O
fractions	O
(	O
11	O
of	O
the	O
33	O
)	O
.	O

Immunostaining	O
showed	O
the	O
estrogen	B-cell_type
receptor-positive	I-cell_type
cells	I-cell_type
to	O
be	O
the	O
macrophage-like	B-cell_type
synoviocytes	I-cell_type
and	O
the	O
CD8+	B-cell_line
,	O
CD29+	B-cell_line
T	I-cell_line
cells	I-cell_line
both	O
in	O
RA	O
and	O
in	O
control	O
synovial	O
tissues	O
.	O

Higher	O
nuclear	O
content	O
of	O
EBS	O
was	O
consistent	O
with	O
more	O
intense	O
nuclear	O
staining	O
of	O
synoviocytes	B-cell_type
and	O
T	B-cell_type
cells	I-cell_type
.	O

CONCLUSION	O
.	O

It	O
is	O
conceivable	O
that	O
the	O
immunomodulatory	O
activity	O
exerted	O
by	O
estrogens	O
is	O
at	O
least	O
partly	O
mediated	O
through	O
their	O
interaction	O
with	O
EBS	O
that	O
are	O
present	O
on	O
macrophage-like	B-cell_type
synoviocytes	I-cell_type
,	O
functioning	O
as	O
antigen-processing	B-cell_type
and	I-cell_type
antigen-presenting	I-cell_type
cells	I-cell_type
,	O
and	O
on	O
antigen-experienced	B-cell_line
(	I-cell_line
memory	I-cell_line
)	I-cell_line
CD8+	I-cell_line
T	I-cell_line
lymphocytes	I-cell_line
(	O
CD29+	B-cell_line
,	O
CD45RO+	B-cell_line