Role of CSBP/p38/RK
cytokine
John
signaling
p38,
Beecham
or
RK,
has
been
laboratories
kinase
via
observed
by a wide
cal stress
and
proinflammatory
independently
cell systems
of stimuLi,
such
necrosis
tokine
mined
biosynthesis
inhibitors
to be
potent
and
factor.
signaling
now
been
inhibitors
cydeterof
These
inhibitors
will
involved
in inflam-
matory
responses.
In particular,
for the first
time
a
definitive
signal
transduction
pathway
can
be prescribed
to the action
of lipopolysaccharide
in cytokme production
in macrophages.
J. Leukoc.
Biol.
59:
152-157;
Key
i996.
Words:
signal
transduction
.
protein
phosphorylation
translation
regulation
.
cytokine-suppressive
drugs
kinase
inhibitor
anti-inflammatory
knowledge
in IL-i
Cells
of the macrophage
or monocytic
lineage
respond
to
external
stimuli
to produce
a large array of proteins,
many
including
secreted
the
( IL-i)
and
growth
factors
also act
receptors
necrosis
[1].
in an
are
most
was
found
to be strongly
rect
result
of LPS
protein
LPS
152
mediators
Journal
has
long
the
is known
cytokines
Leukocyte
about
interact
a variety
of
cytokines
can
(MAP)
was
known
as one
[3].
Identifica-
and the subwith a cell
molecular
events.
been known
to be
Even
though
much
their respective
receptors
and
with their receptors,
there
is
Biology
Volume
59,
February
1996
has
been
phosphoryproposed
by a
also
[13,
14]. A prominent
CD14-transfected
tyrosine
kinases,
phosphorylated
p42/p44
also
known
MAP kinase
to be implicated
observed
that
IL-i
protein
iso70Z B cells
as a di-
mitogen-activated
as Erks,
and
their
kinase
(MEK)
were
in IL-i
signaling
and
TNF
are
unique
kinases
to cytokine
stimulation
because
are also able to phosphorylate
terizing
the activation
[i9] found that it was
of MAPKAP
not stimulated
but
such
by
other
stimuli
particularly
in
[13,
We have
of regulation
been interested
of cytokine
approach
was
cytokine-suppressive
were
examined.
17,
The
in
LPS-
the molecular
in monocytes.
in the
a series
earliest
have
basis
Our
literature
of small
compounds
also
there
has been a paucity
of action
[2i]. We were
an
also
phos-
i8].
inhibition
activity
were
an increasing
number
inhibitors
et al.
factor
through
production
14,
survey
reports
effects
of
showed
cytokine
synthesis
cocorticoids
[20]. Recently,
molecular-weight
stress
in defining
biosynthesis
to
Rouse
growth
of recent
reports
serine/threonine
cytokine
macrophages
initial
where
as chemical
regulating
stimulated
poIt should
is not
a variety
of other
Hsp27.
In charac-
kinase-2,
by nerve
pathway.
A number
for both tyrosine
and
up-
among
[i5]
It
been
that
the gluof low-
described,
of information
encouraged
on their
by a report
of
provided
a receptor
basis
little
definitive
data has
[7].
transduction
tent inducers
of Hsp27
phosphorylation
[i6-i8].
be noted,
however,
that
Hsp27
phosphorylation
although
mechanism
of the cytokine
[2]. Bacterial
(LBP)
complex
post receptor
also have long
of inflammation
of
been
binding
protein
of the LPS/LBP
TNF
these
of macrophages
termed
CD14
[4-6].
However,
come forth delineating
Similarly,
IL-i
and
that
interleukin-l
and
fashion,
as many
on these
cells
activators
surface
molecule
for LPS action
(TNF),
signal
protein
signaling
stimulation.
stream
activator,
the first kinases
molecules
active,
cytokines,
factor
(LPS)
potent
information
how these
pharmacologically
It is interesting
autocrine
expressed
tion of a specific
sequent
interaction
potent
are
proinflammatory
tumor
lipopolysaccharide
the
and
TNF
respectively
LPS-stimulated
Erk-independent
suggest
a role
I NTRODUCTION
are
their
of serine/threonine
and
Han et al.,
hated from
phorylation
of which
regarding
number
of investigators
[8-12].
The first definitive
report
suggesting
that LPS signaling
also involves
protein
phosphorylation
came
from the studies
of Weinstein
et al. and
or
1 and
CSAIDTM
have
selective
activity.
pathways
in LPS and
19406
mechanisms.
The involvement
lation
novel
been
upon
stimulation
as physicochemi-
Furthermore,
Pennsylvania
limited
by
oftbis
has
with
Iipopolysaccharide
such as interleukin-
tumor
kinase
of Prussia,
mitogen-activated
termed
CSBP,
identified
treatment
cytokines
CSBP/p38/RK
help to dissect
King
recently.
Activation
dual
phosphorylation
in different
spectrum
kinase
R. Young
Pharmaceuticals,
A new
member
of the
kinase
family,
alternatively
several
protein
response
mechanisms
C. Lee and Peter
SmithKline
Abstract:
protein
stress
Abbreviations:
IL-i,
.
pressive
L
vated
r,
TNF,
.
L
kinase;
iNK,
necrosis
TM
.
protein;
CSBP,
drugs;
regulated
tumor
.
oinciing
anti-inflammatory
extracellular
1
CSAID
c-jun
kinase;
I
binding
MAP,
factor;
,
LPS,
cytoKine-supprotein;
Erk,
mitogen-acti.
protein
Reprint
SmithKline
PA
interleukin-i;
.
Iipopolysaccnaricle;
requests:
Beecham
John
C. Lee,
Department
of Cellular
P.O.
Pharmaceuticals,
Box
1539,
Biochemistry;
King
of Prussia,
19406.
Received
November
September
21,
1995.
18,
1995;
revised
November
20,
1995;
accepted
suggesting
lipoxygenase
higher
than
inhibited
that
inhibitors,
observed
IL-i
at
for their
production
in
human
inhibition,
also
monocytes
testing
a series
of novel,
imidazole-containing
matory
compounds
that had previously
been
inhibitors
of eicosanoid
metabolism
in enzyme
a few demonstrated
IL-i
synthesis
there was no correlation
between
SK&F
concentrations
enzyme
[22].
In
anti-inflamshown
to be
assays
[23],
inhibition
activity,
cytokine
suppression
potency
of eicosanoid
inhibition.
One compound,
SK&F
86002,
did consistently
strate
cytokine-suppressive
activity
(ICso
1.3
but
and
demon± 1 tM)
86002
subsequently
inflammatory
relatively
to
reveal
the
with
IL-i
system
stimuli
and
was
target
observed
cells.
with
Optimal
a number
of different
inhibition
was
unique
of TNF production
the latter
study,
shock-related
holding
even
iting
observed
bone
related
activities
of
exhibited
[26]. Most
arthritis
were
models
for anti-
systems
inhibitors
that were
in order
cytokine-suppressive
significant
activity
in many
notable
is its effectiveness
model
[27] and inhibition
in the endotoxic
the reduction
in
survival
analogues
of animal
We chose
model
to cyclooxygenase
agents.
SK&F
86002
such in vivo studies
in the collagen-induced
correlated
structurally
in a number
activity.
insensitive
[24]. This compound,
when
tested
at its IC50 for cytokine
inhibition,
had no appreciable
effect
on DNA,
RNA,
or
protein
synthesis.
Furthermore,
its inhibitory
activity
on
production
and
tested
and
shock
plasma
model
level
protection
[28]. In
of TNF
from
endotoxic
mortality.
Equally
exciting
and
greater
promise
is its effectiveness
resorption
in a rat fetal
long-bone
perhaps
inhib-
in
organ
culture
[29].
To further
understand
CSAID
act at
a variety
of biochemical
apthese
compounds
act predomiwith limited
effect
on cytokine
prompted
mRNA
blot
naming
drugs
them
cytokine-suppressive
anti-in-
[30-32].
we proceeded
compounds
the site
of action
using
proaches.
We found
that
nantly
at the protein
level
our
level,
the
the
flammatory
molecular
how
when
the cells
were pretreated
or treated
early
(<2
h) in
the induction
phase
of IL-i expression.
Both IL-i isoforms
were inhibited
to a similar
extent,
as was TNF production
[25]. This unique
activity
associated
with these
molecules
Western
to systematically
analysis
confirmed
intracellular
levels
of both cytokines
duced
in treated
cells
and were not
(CSAIDTM).
dissect
that
the
were significantly
reparalleled
by similar
changes
in the respective
mRNAs.
No significant
effect
was observed
in transcript
size, protein
half-life,
or protein
secretion
(P. R. Young,
P. C. McDonnell,
J. T. Laydon,
W.
Prichett,
U. Prabhakar,
Lee,
unpublished
analysis
suggested
[32,
SK&F
86055
SK&F
86002
33,
and
increasingly
mechanism
Young
et al.,
the molecular
which
we call
implication
for
IDENTIFICATION
TARGET
The
human
source
identify
monocytic
tam
suitable
showed
that
(Fig.
SB 203580
1. Structures
the
86002
highly
70
and
kinase
86055
was
Ci/mmol)
was
effect
activity.
used
loclinated
the
protein.
SB
binding
as well
as the
SK&F
203580
kinase
86002
SB
the
SB 206718
is a potent
assay.
discussed
86055
were
of cytokine
Tritiated
in developing
affinity.
in studies
SK&F
of inhibition
inactive.
of binding
target
used
its regioisomer
specific
binding,
SK&F
ofcompounds
and
data).
it
It became
elucidate
the
was
essential
Identification
of the target
(CSAID
binding
protein),
cytokine
OF THE
J. C.
production
will
be
re-
MOLECULAR
was
the
202190
binding
was
and
in the
to show
production,
ligand
active
compound
(radiospecific
assay
used
and
and
activity
determination
to photoaffinity
specific
review.
employed
inhibitor
label
in the
cell
line,
THP.i,
was
used
as
a
of target
protein
because
previously
it had been
to give a response
similar
to human
monocytes.
To
the molecular
target,
it was first necessary
to oh-
shown
N
Fig.
unpublished
target.
CSBP
and
polysome
gradient
prior to elongation
SB 206718
SB 202190
SK&F
J. Adams,
clear
that
in order
to further
of action
of these
compounds,
to identify
molecule,
and
its
viewed.
D. Dunnington,
data).
In addition,
that a block occurred
i)
radiohigands.
the uptake
by THP.1
of
cells
Cellular
uptake
the radioligand,
was
experiments
SB 202190
time-dependent,
saturable,
and
specific.
Upon
subcellular
fractionation
of radioligand-loaded
THP.1
cells,
the predominant
subcellular
site
of accumulation
of the radioactivity
was the cytosol.
A
simple
and reliable
developed
to separate
a Sephadex
G-10
competitive
free from
minicolumn
drug
binding
bound
radioligand
assay
was
by using
[34].
The binding
activity
was again
specific,
saturable,
and
reversible.
Scatchard
analysis
revealed
that the average
dissociation
constant
of binding
was 50 nM with
a single
binding
site
Lee
and
et (11. CSBP
was
of low
kinase
iii
abundance
LPS
and
(-5
cytokiiie
pmol/mg).
signaling
To
153
verify that
inhibition
binding
to this target
of cytokine
synthesis,
analogues
representing
protein
was
a number
various
for cytokine
biosynthesis
petitive
binding
assay
potencies
inhibition
Cytokine
were used
inhibition
highly
with binding
coefficient
of 0.9
to THP. 1 cytosol,
[34].
In addition,
bioassay,
effect
the
drug
in the
in the
including
tivities
and/or
are
inhibitory
activity
cyclooxygenase,
compete
in the
Lee,
unpublished
strong
evidence
directly
CSAID
those
that
had
binding
linked
to cytokine
compounds.
biosynthesis
ac-
lw
Fig.
2.
Maps
structure
Four
almost
full
length
encompassed
except
the
for
clones
5’ and
separate
the
3’
of CSBP.
UTR
(solid
clones
spanning
both
5’ and
open
reading
complete
75
site
as shown.
At the
box)
eDNA
internal
polyadenylation
and
were
top
is shown
open
obtained
frame
nt (boxed).
Clones
frame
N5
(CSBP2)
with
3’ regions.
the overall
reading
Clone
N13
was
identical
and
N7/N9
used
(open
being
(CSBP1)
to N5
an
alternate
by the
202i90
cates
AND SEQUENCE
OF CSBP
radioligand.
the CSBPs
that
mRNA
PURIFICATION
DETERMINATION
of cDNA
with
box).
failed
to
and J. C.
provide
protein
is
inhibition
l.&w
l’--
N5
cytokine
biosynthesis
A, dexamethasone,
and
lipoxygenase
inhibitors,
binding
assay
(J. M. Blumenthal
observations).
These
results
that ligand
binding
by the target
I
I
N7/N9
N13
assay
anti-inflammatory
implicated
to have
such as cyclosporin
3.8kb
EcoRl
BPO1/BPO2
with a correlation
as observed
in the
was regioselective
as well,
SK&F
86002
versus
SK&F
86055
(Fig.
1). A large
number
of structurally
unrelated
compounds,
:
bioassay
in the comcorrelated
competitive
?
0
responsible
for
of structural
of ‘-4.2
and FISH
coded
by
Examination
are encoded
kb.
analysis
a single
Analysis
of Northern
by a fairly
blots
mdiubiquitous
of human-rodent
indicate
that
gene
mapped
cell
fusions
both CSBP
forms
to chromosome
are en6p2i.2
[35].
The radioligand
binding
assay,
although
useful
in establishing
the existence
of the binding
protein,
proved
to be
A search
of genetic
databases
quences
showed
that the genes
inadequate
kinase
family
[36, 37].
identity
with the yeast
Mpk2
[i9], and shares
to support
more
the protein.
A photoaffinity
i) was employed
to detect
than
a 20-fold
radioligand,
the target
purification
SB
protein.
206718
Upon
of
(Fig.
ultra-
violet
activation,
a single
discrete
radioactivity-crosslinked
protein
with an apparent
molecular
mass of 42 kDa,
p1 of 5.2 was specifically
detected
[34].
The molecular
MAP kinase
lar regulated
mass
kinase
is similar
to that
determined
previously
by size
the
exclu-
sion chromatography
in the binding
assay,
suggesting
that
CSBP
is monomeric.
Competition
of radiophotoaffinity
cross-linking
with
various
structural
analogues
demonstrated
identical
rank-order
potency
to the binding
assay.
A combination
of the binding
assay and photoaffinity
labeling were used to purify
the protein
and, after chemical
and
proteolytic
were
fragmentation,
two
unique
peptide
sequences
obtained.
CLONING
Using
based
AND
EXPRESSION
two reverse-translated
on these
peptide
were
cloned
from
OF CSBP
KINASE
(Fig.
due
in a 75-nucleotide
region
the DNA
and amino
acid
2).
Sequencing
to an alternative
of genomic
splice
within
the coding
region,
identity
was only 43%
confirms
(McDonnell
et al.,
that
this
unpublished
results).
The two proteins
were named
CSBP1
and CSBP2
and, when
expressed
in Escherichia
coli, could
cross-link
to radiophotoaffinity
compound
and bind
to the [3H]SB
154
Journal
of
Leukocyte
Biology
Volume
59,
February
1996
(JNK)
transcription
named
alternatively
Woolf,
factor
for its ability
spliced
CSBP1
P. C. McDonnell,
data).
CSBPs
required
(extracelluand activate
Elk-i,
and
c-jun
to phosphorylate
is also
P. R.
and
found
Young,
yeast
kinase
with
cells
kinase
mutant
loop
of this
tyrosine
the
N
confirmed
sites:
that
TGY
TPY for iNKs.
It is now
important
in determining
MAP
kinase
kinase.
mouse
unpublished
(D.
regions
and
MAP ki-
proximal
to the
family,
phosphoby a dual-speci-
is required
for kinase
activation.
CSBP2
expressed
in mammalian
phosphorylation
and Yi82
of CSBP
is essential
for its
unpublished
data].
Similar
results
have
the rodent
and Xenopus
homologues
[i7,
ing that CSBP
differs
from the Erks and
of this loop and the intervening
residue
kinases
kinases
in
retain
all of the i 1 conserved
kinase
residues,
and contain
the characteristic
phosphorylation
is
Erks
terminus
of c-jun.
leading
to its transcription
activation.
Alignment
of these
closely
related
sequences
(Fig.
3)
shows the evolutionary
relationship
of the homologues.
The
and
colony-stimu-
DNA
also shares
51% amino
acid
[38], 88% with the Xenopus
identity
with the murine
p38
[39]. Closely
related
are the
kinases),
which
phosphorylate
serum-responsive
ficity MAP
Experiments
hating
factor-stimulated
human
monocyte
library.
Two of
these
cDNAs
contained
a complete
open reading
frame
of
360 amino
acids containing
the two peptide
sequences,
but
differed
where
CSBP2
HOG1
99.8%
nase
TXY
motif
in the regulatory
active
site. In all known
members
rylation
of both the threonine
and
degenerate
oligonucleotides
sequences,
several
CSBP
cDNAs
a granulocyte-macrophage
with the two CSBP
Sewere members
of the MAP
for
CSBP,
on
Ti80
activation
[39a;
been reported
for
40]. It is interestJNKs
in the size
between
the two
TEY
for
Erk,
and
clear
that these
differences
are
the specificity
of the activating
Thus,
have been
identified
(MEKi,
MEK2
for
unique
activating
for each of these
erk,
MKK3
for
MAP
three
CSBP,
kinase
MAP
and
,__l- human
i_I
I
3.
I
1-xenopusmpk2
H
yeast
Phylogenetic
family.
Human
kinase)
and
relationship
CSBP2
among
shared
to CSBP1.
The
the
highest
most
TNF-a
csbpl
synthesis
jnkl
phatase
erk2
increase
phorylation
ofthe
MAP
murine
kinase
CSBP2
(p38
erk.
for
CSBP
and
iNK)
[4i-43],
and
there
is
stresses,
44-46],
whereas
differentiation
IL-i,
the
TNF,
erks
OF CSBP
KINASE
CSBP
kinase
activity
can
tion of the generic
substrate
activated
chemical,
in monocytes
osmotic,
and
CSAID
[i#{244}-i9, 34,
by mitogenic
best-studied
CSBP
iM [i8; J. C. Lee,
CSBP
inhibitor
S. Kassis,
observations].
The
such
as okadaic
[50,
cytokine
to and
tyrosine
indicating
a high
203580
does
not
TNF
level,
and
phosphorylation
mar et al, unpublished
communication).
The
lowed
further
delineation
by upstream
data, and
availability
of the
kinase
is
activity
of
NF-KB-associated
in
for the first time
TNF is mediated
cascade
that
sion of these
constructs
suggest
through
evidence
that
proteins
or
direct
that the translation
of IL-i
an independent
phosphorylation
and
is absolutely
required
for the ultimate
expresproteins.
Previous
studies
with reporter
gene
in which
various
of -0.22
unpub-
at up
deletions
have
been
made
and
LPSiIL-1
strs
to iOO
kinases
in vivo
V
(Ku-
‘i00
Thr GUy Tyr
et al, personal
inhibitor
has al-
phosphorylation
cascade
downstream
from CSBP
activation
in vivo. For example,
stress-stimulated
cells,
the compound
inhibits
activation
MAPKAP
kinase-2,
a physiological
substrate
of CSBP,
its phosphorylation
of Hsp27
[18] (Fig.
4).
The
also
is stimulated
is some
of specificity.
ImporCSBP
activation
or
Saklatvala
of this
ROLE OF CSBP KINASE
IN THE
OF CYTOKINE
BIOSYNTHESIS
synthesis
there
as observed
in the
SB 203580,
the
(tested
degree
inhibit
phosIL-i
CSBP.
activation,
at the transcriptional
effectively
by disrupting
the
balance,
stimulate
inactivating
monocyte
which
to be
phos-
51]. The target
of the phosphatase
induction
cascade
may be at a
j.tM) did not inhibit
a range
of other
protein
kinases
tested
in vitro
including
the MAP kinase
homologues
JNK and
Erk as well as the upstream
activating
kinase(s)
of CSBP
[i8, 47; Cohen
et al., and Saklatvala
et al., personal
communication]
tantly,
SB
acid,
phosphorylation
dephosphorylation
toxicity
likely
with
phosphorylation
of the c-jun
component
of AP-i
[45, 53].
Whether
this involves
the MAP kinases,
such as CSBP
or
JNK in particular,
has not been clearly
determined.
It has also been established
that TNF-a
and IL-iD
can
be regulated
at the translational
level [54-56].
Our studies
(Fig. 1) has an IC50
and J. M. Blumenthal,
compound
lethal
this
block
phosphorylation
by LPS and in various
cell lines by
heat stress
as well as by cytokines.
inhibit
LPS-induced
to
44,
INHIBITOR
rank-order
potency
and binding
assays.
prevent
of
appears
and
be detected
by the phosphorylamyehin
basic
protein.
CSBP
compounds
vitro with the same
cytokine
inhibition
hished
LPS)
activated
stimuli.
PROFILE
The
and
are
Inhibition
phosphotylation
is also
treatment
of macrophages
protein
and
leading
[48].
inhibitors
this may be in part mediated
by the transcription
factors
nuclear
factor KB (NF-KB)
and apotinin-i
(AP-i)
[52]. For
both IL-i
and TNF,
phosphorylation
can play a regulatory
role in transcriptional
activation
either
through
indirect
evidence
for additional
isoforms.
This
is consistent
with
the finding
that both
CSBP
and JNK
are activated
by a
variety
of stress
and cytokine
signals
(e.g., heat, chemical,
osmotic
and
production
in the
Upon
kinases
kinase
inhibitors,
or TNF
inhibitor
step
MKK4/SEKi
MAP
tyrosine
[49]. Serine/threonine
involved
because
HOC1
is the
the
by
human
with
member
including
human
members
identity
distant
i4]
pathway
L__ human
I
Fig.
csbp2
Lmunnep3g
0
in
of
and
Ser/Thr
(PKA
I
REGULATION
discovery
of the role of CSBP
in cytokine
production
helps
to shed some light on our understanding
of the
responses
of monocytes
and
macrophages
to endotoxin.
Several
studies
have implicated
protein
phosphorylation
in
these
responses.
For example,
a number
of intracellular
substrates
appear
to undergo
tyrosine
phosphorylation
[i3,
I HSP27]
Fig.
4
CSBP
cascade
as
cellular
is a stress-response
well
stresses,
which
in
CSBP
activation.
which
prevents
h0S)h0rylati0n
turn
as
kinase
downstream
and
LPS
activate
phosphorylate
CSBP
CSBP
activity
phosphorylation
of Hsp
Lee et a!. CSBP
with
targets.
CSBP
at
its
own
distinct
activation
Pro-inflammatory
Thr
is inhibited
of MAPKAP
cytokines,
upstream
180
and
kinases
Tyr
182
specifically
kinase-2
MKK3/4,
resulting
in
by SB 203580,
and,
subsequently,
27.
kinase
in LPS
and
cytoidne
signaling
155
transfected
into macrophage
cell lines have suggested
that
the regulation
of TNF translation
is mediated
through
an
AUUUA
repeated
motif
in the 3’UTR
of the TNF mRNA
[57]. Deletion
of this region
leads
to constitutive
synthesis
both
in
macrophage
and
non-macrophage
cell
[58-60]
and in transgenic
animals
[oi]. With
porter
constructs,
we have shown
that inhibition
gene expression
by the CSAID
cytokine
inhibitors
ated
data).
through
These
and
translation,
but
of a protein
similar
reof reporter
is medi-
through
the novel
ered
might
cause
TNF-cz
and
Although
possible
complex
that
sitting
phosphorylation
release
of
cascade
translational
site
in response
5.
of
in the production
stimuli
mRNA
borne
to various
macrophages,
the ubiquitous
presence
of CSBP
[34]
suggests
a role in other
cell types.
This
is
out by the example
of IL-i
induction
of IL-8
in
iii
cells,
which
inhibitors
that these
inhibitors
duci ng i nflammatory
The
role of these
production
can
[30].
also be inhibited
by
It is therefore
likely
block
a number
of pathways
for procytok i nes.
cytokine
inhibitors
in blocking
the
of inflammatory
cytokines
limited
effect
on T or B cell
production
of cytokines
such
contrasts
responses
as IL-2
with
[30] involving
and IL-4.
By
1.
2.
3.
4.
5.
6.
their
7.
the
com-
8.
1 1.
inhibited
by SB
12.
13.
14.
15.
kDaTNF
16.
secretion
and
Fig.
5.
Model
to translational
by
and
156
We
In
the
direct
uptake
of the
Journal
is both
propose
mRNA
and
cytokine
biosynthesis
in the
inhibitor,
of Leukocyte
active
and
ofthe
with
one
17.
leading
in monocytes.
presence
event(s)
Upon
mRNA
translational
of the
components
resulting
in release
polynbosomes.
Biology
Volume
59,
18.
translation
inhibitor,
an inactive
or more
phosphorylation
into
signaling
transcription
associates
of the
or indirect
IL-i
increased
that
sequences
absence
in LPS
of cytokine
there
3’ AU rich
undergoes
and
LPS,
TNF.
containing
complex.
of CSBP
regulation
stimulation
oflL-i
role
animal
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a novel
therapeutic
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