|
|
 Previous Article | Table of Contents | Next Article 
Blood, Vol. 93 No. 5 (March 1), 1999:
pp. 1456-1463
Interleukin-10 Inhibits Expression of Both Interferon  - and
Interferon  - Induced Genes by Suppressing Tyrosine
Phosphorylation of STAT1
By
Satoshi Ito,
Parswa Ansari,
Minoru Sakatsume,
Harold Dickensheets,
Nancy Vazquez,
Raymond P. Donnelly,
Andrew C. Larner, and
David S. Finbloom
From Division of Cytokine Biology, Center for Biologics Evaluation
and Research, Food and Drug Administration, Bethesda, MD.
 |
ABSTRACT |
Interleukin-10 (IL-10) helps maintain polarized T-helper cells in a
T-helper lymphocyte 2 (Th2) phenotype. Part of this
process involves the prevention of the development of Th1 cells, which are a primary source of interferon  (IFN), a potent activator of
monocytes and an inhibitor of Th2 proliferation. Because monocytes and
macrophages are important mediators of Th1-type responses, such as
delayed-type hypersensitivity, we sought to determine if IL-10 could
directly mediate inhibition of IFN- and IFN -induced gene
expression in these cells. Highly purified monocytes were incubated
with IL-10 for 60 to 90 minutes before the addition of IFN or
IFN. IL-10 preincubation resulted in the inhibition of gene
expression for several IFN-induced genes, such as IP-10, ISG54, and
intercellular adhesion molecule-1. The reduction in gene expression
resulted from the ability of IL-10 to suppress IFN-induced assembly of
signal transducer and activator of transcription (STAT) factors to
specific promoter motifs on IFN- and IFN-inducible genes. This
was accomplished by preventing the IFN-induced tyrosine phosphorylation
of STAT1, a component of both IFN- and IFN-induced DNA binding
complexes. Therefore, IL-10 can directly inhibit STAT-dependent early
response gene expression induced by both IFN and IFN in monocytes
by suppressing the tyrosine phosphorylation of STAT1. This may occur
through the ability of IL-10 to induce expression of the gene,
suppressor of cytokine signaling 3 (SOCS3).
This is a US government work. There are no restrictions on its use.
| |
INTRODUCTION |
INTERFERON  (IFN) has been shown to
be therapeutically effective in the treatment of certain types of
leukemias and lymphomas1,2 and in some solid tumors such as
melanoma.3 Although not used as frequently as IFN,
IFN has been shown to have some activity in renal cell carcinoma and
is a major secretory product of T cells stimulated with interleukin-12
(IL-12).4 Obviously, not all tumors respond to IFNs, and
the effects of IFNs on cells can be modulated by many different agents.
A cytoplasmic protein is present in tumor cells that can block the
formation of DNA-binding proteins that recognize the IFN-stimulated
response element of IFN-induced genes.5 T-cell cytokines
such as granulocyte-macrophage colony-stimulating factor (GM-CSF) can
also effectively block the ability of IFNs to activate monocytes, cells
known for their ability to mediate cytotoxic effects against tumor
cells.6 Therefore, the effects that monocytes exert in any
environment depend to a large extent on the type of T-cell cytokines
present at the time they interact with other cells.
Immune responses mediated by monocytes and activated T cells can be
divided into two types, based on the expression of groups of
T-cell-derived cytokines.7 Although most clearly defined in murine models of parasitic diseases, such as
Leishmania,8 the ability to classify certain human diseases
into one polarized type or another is a useful tool to learn more
regarding the immunologic basis of that particular disease. A cellular
immune response dominated by a T-helper lymphocyte 1 (Th1)
infiltration, in which IFN and IL-2 predominate, is observed in
diseases such as rheumatoid arthritis9 or inflammatory
bowel disease.10 This response generally evokes a
delayed-type hypersensitivity reaction. Immune responses regulated by
T-helper lymphocyte 2 (Th2) cells result in the secretion of IL-4 and
IL-10. This response is important for humoral immunity and mast cell
development seen in such diseases as hyperimmunoglobulin E secretion syndrome.
It has been appreciated that cytokines secreted by Th2 cells can
dramatically modulate the responses of cells involved in a Th1 immune
response. For example, IL-4 can modulate the expression of early
response genes induced by IFN and IFN.11 Likewise, IFN can affect the response of cells to IL-4.12 Whereas
it is known that IL-10 can effectively block lipopolysaccharide
(LPS) activation of monocytes,13 its effects
on direct IFN activation of genes are poorly understood. IFNs and IL-10
bind to their cognate receptors and initiate a signal that results in
the activation of Janus kinase (Jak) and signal transducer and
activator of transcription (STAT) proteins, leading to transcription of
early response genes.14 IL-10 activates Tyk2, Jak1, STAT1,
and STAT3.15 IFN activates Tyk2, Jak1, STAT1, STAT2,
STAT3, and STAT5. IFN activates Jak1, Jak2, STAT1, and, in some cell
lines, STAT5.16 Although there is some overlap among the
proteins activated by these three cytokines, there is no clear
explanation from this information as to why one IFN or one IL should
inhibit the action of others. We hypothesized several years ago that
GM-CSF could inhibit the expression of IFN-induced genes by
activating a new STAT protein that bound to the same activation
sequence (GAS) element as IFN-induced STAT1 and thus prevent the
induction of the FcRI gene.17 This protein was
subsequently determined to be STAT5. Although it is well established
that IL-10 can suppress LPS-induced responses, which are by and large
driven by NF B, its direct effect on the ability of IFN and IFN
to activate cells by an NFB-independent mechanism is virtually
unexplored. In this report, we describe the ability of IL-10 to inhibit
the expression of specific sets of IFN response genes. We show that
IL-10 can suppress IFN-induced expression of several genes by
inhibiting STAT1 tyrosine phosphorylation. In addition, we have gone on
to show that IL-10 markedly upregulates expression of the SOCS3 gene,
which is known to suppress cytokine activation.
| |
MATERIALS AND METHODS |
Monocytes.
Human peripheral blood leukocytes from healthy donors were separated
into a mononuclear cell fraction and then were subjected to
countercurrent centrifugal elutriation. The fractions enriched for
monocytes were cultured in Dulbecco's modified Eagle medium (DMEM;
GIBCO, Grand Island, NY) without serum for 45 to 60 minutes in 4-well
tissue culture plates until adherent, washed, and then incubated with
DMEM with 10% fetal bovine serum (Hyclone, Logan, UT) and gentamicin
(50 µg/mL). Cells prepared in this manner are greater than 95%
monocytes by morphology and staining for anti-CD14. Cells were
incubated with recombinant human IL-10 (rhuIL-10; Schering-Plough, Madison, NJ) for 60 to 90 minutes before the incubation with IFN (Roferon; Roche, Nutley, NJ) or IFN (Genentech, South San Francisco, CA) for the specified times.
RNAse protection assay.
Steady-state levels of mRNA induced by either IFN or IFN were
measured by RNAse protection assay, as previously
described.18 For the assay, probes were constructed such
that each lane of the gel contained several protected fragments.
-Inducible gene-10 (IP-10), a chemokine with
anti-angiogenesis properties, was linearized with Pvu II, which
yielded a protected fragment of 360 bp.19 ISG54, a gene
induced by IFN (and in monocytes by IFN), was linearized with
Pvu II, and this yielded a protected fragment of 367 bp.20 Guanylate binding protein (GBP; an IFN- and
IFN-induced gene) was linearized by Asp718.21 For an
internal control for the amount of RNA loaded onto the gel,
glyceraldehyde-3-phosphate dehydrogenase (GAPDH; linearized with Sau3a)
was used and had a protected fragment of 300 bp.22 All
hybridizations were performed overnight at 56°C. The RNA was
subjected to digestion with RNAse T1, followed by separation on a 6%
polyacrylamide/urea sequencing gel, and analyzed by phosphorimager
analysis and autoradiography.
Northern analysis.
Total RNA was isolated from cultured monocytes by the acid guanidinium
thiocyanate-phenol-chloroform extraction method, as previously
described.23 RNA precipitates were pelleted by
microcentrifugation and redissolved in 100 µL of diethylpyrocarbonate
(DEPC)-treated sterile water. Equivalent amounts of RNA
(10 µg/lane) were size-fractionated by electrophoresis in 1% agarose
gels containing 0.66 mol/L formaldehyde. The RNA was then blotted by
overnight capillary transfer onto Nytran membranes (Schleicher & Schuell, Keene, NH) and cross-linked by exposure to UV light. The
membranes were then prehybridized, hybridized, and washed according to
standard procedures. The SOCS3 probe was a 681-bp Mlu I
fragment of the full-length murine SOCS3 cDNA,24 kindly
provided by Dr Douglas Hilton (Walter and Eliza Hall Institute,
Parkville, Australia). Gel-purified insert DNA was radiolabeled by the
random primer method of Feinberg and Vogelstein25 to a
specific activity of 108 cpm/µg or greater.
Electrophoretic mobility assays (EMSA).
EMSA using the response region (GRR) of the FcRI receptor were
performed using either whole-cell extracts or nuclear extracts, as
described previously.26,27 This element is similar to the GAS element found within many IFN-induced genes. This probe was also
used for the study of IFN-induced genes, in addition to the
IFN-stimulated response element (ISRE) from the ISG15 gene, which is
recognized by the ISGF3 complex and is specific for IFN-induced genes.11
Immunoprecipitations.
After treatment with IL-10 and/or IFNs, monocytes were
solubilized with 1% TX100-containing buffer, incubated on ice for 15 minutes, and then centrifuged for 13,000g for 12 minutes to
generate a postnuclear extract as described.15 The
postnuclear extracts were then incubated with anti-STAT1 antibodies.
The immunoprecipitates were separated by sodium dodecyl
sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gels, transferred
to a polyvinylidene difluoride membrane, and immunoblotted with
antiphosphotyrosine. Alternately, some extracts were analyzed directly
by SDS-PAGE and immunoblotting with antiphosphotyrosine-STAT1 (New
England Biolabs, Beverly, MA). Immunoblots were developed using either
horseradish peroxidase or enhanced chemiluminescence (ECL) or nitroblue
tetrazolium chemistry, respectively.
| |
RESULTS |
We initially addressed the question of which early response genes
induced by IFNs might be inhibited by IL-10. Studies with STAT1-deficient mice showed that induction of IP-10 is dependent on
STAT1 activation.28 The IP-10 gene encodes a chemokine of the cysteine-x-cysteine (CXC) type that is rapidly
activated by IFN and IFN. Recent data have shown that the IP-10
protein has strong anti-angiogenic properties and, in addition to
inhibiting the development of carcinogenic tumors in several mouse
models, has profound effects on chronic inflammatory
conditions.29-31 We initially examined the effect of IL-10
on the induction of IP-10 in monocytes exposed to IFN (Fig
1A). We observed rapid induction of IP-10
expression that was markedly inhibited by IL-10 at all concentrations
of IFN tested. For some individuals (such as the one shown), nearly
complete inhibition could be observed. Although the number of donors
were only two per group, it was clear from the autoradiograph that at
higher concentrations of IFN, there was less inhibition by IL-10.
IFN-stimulated IP-10 RNA expression was inhibited by pretreatment of
monocytes with IL-10 (Fig 1B). At virtually all doses of IFN, over a
range that occupies approximately 10% to 100% of the receptors for
IFN, IL-10 suppressed IFN-induced IP-10 RNA expression by 59% to
68%. However, at the lower concentrations of IFN, inhibition tended
to be greater. We went on to confirm that there was no change in
receptor number of affinity by directly measuring the number of IFN
receptors on monocytes after IL-10 treatment (data not shown).
View larger version (67K):
[in this window]
[in a new window]
| Fig 1.
IL-10 inhibits the induction of IP-10 and ISG54 by IFN
and IFN. Monocytes were treated with 10 ng/mL of IL-10 for 60 minutes before treatment with the indicated doses of (A) IFN or (B)
IFN for 90 minutes. RNA was isolated from the cells, and RNAse
protection assays were performed. The arrows indicate the protected
fragment corresponding to IP-10 and the control gene, GAPDH. IL-10
inhibits IFN-induced expression of ISG54 by (C) IFN or (D) IFN.
The arrows indicate the protected gene fragment for ISG54 and the
control gene, GAPDH. For all RNAse protection assays, the ratio of the
values for the IFN-induced gene divided by those for the control
(GAPDH) gene was computed, and for cells that had been pretreated with
IL-10 followed by IFN. The values were calculated from Phospho-Imager
data analysis. The percentage of inhibition was calculated by
subtracting the values for cells treated with IL-10 from values for
IFN-treated cells and dividing this number by the values for the cells
treated with IL-10 and IFN. The values under the gel represent the mean
of 2 to 6 experiments ± standard deviation at the various doses.
|
|
In addition to IP-10, we also examined IFN activation of ISG54 (Fig
1C). Although ISG54 was one of the first genes recognized as an
IFN-inducible early response gene, virtually no information is
available regarding the function of the protein that it encodes. IFN-stimulated ISG54 is entirely dependent on the assembly of a
complex of transcription factors, including STAT1, STAT2, and p48, that
bind to the ISRE enhancer. Prior incubation of monocytes with IL-10
resulted in IFN-induced inhibition of the accumulation of ISG54 RNA,
compared with incubation with IFN alone (Fig 1C). The observed
inhibition by IL-10 was greater when cells were incubated with
submaximal concentrations of IFN (49% v 17%). There is
some variability, and some donors clearly show inhibition of greater than 60%. Early reports discounted the ability of IFN to induce the
ISG54 gene because of the lack of a GRR sequence within its 5'
regulatory region. However, IFN treatment of peripheral blood monocytes can clearly stimulate the accumulation of ISG54 RNA (Fig 1D).
Incubation of monocytes with IL-10 showed a consistent suppressive
effect on IFN-stimulated ISG54 RNA expression to a similar extent as
that observed for the IP-10 gene. In many donors, the suppression
reached levels of greater than 90% at lower doses of IFN (1 ng/mL,
in this experiment; Fig 1D). These concentrations of IFN are
generally at a level in which receptors are only 10% occupied, but
genes are maximally expressed.27
In addition to IP-10 and ISG54 genes, we examined the ability of IL-10
to suppress the induction of several other IFN-inducible genes. These
genes included ICAM-1 and IRF-1. Although the IRF-1 gene has GAS
elements that drive its transcription by IFN, there was no
perceptible inhibition by IL-10 in monocytes. For ICAM-1, only IFN
was capable of inducing its expression; IFN had no effect (data not
shown). ICAM-1 expression is clearly important for immune cell
adherence and tumor cell recognition.32,33 For those
experiments in which this gene had increased expression of at least
fourfold after exposure to IFN, pretreatment with IL-10 inhibited
ICAM-1 RNA expression by 50% (over a range of IFN concentrations of
0.25 to 10 ng/mL; data not shown). These results are consistent with a
recent report describing the inhibitory effects of IL-10 on
IFN-induced ICAM-1 expression in monocytes.34
Both IFN and IFN activate STATs, which activate gene expression
by binding to ISRE and GAS elements in the promoters of IFN-inducible
genes. IFN essentially activates only STAT1 in human peripheral
blood monocytes, although one group has reported the activation of
STAT5 in the histiocytic lymphoma cell line, U937.35 IFN
can activate several STATs, including STAT1, STAT2, STAT3, STAT4, and
STAT5. Multimers of these STATs bind to ISRE or GAS
enhancers to facilitate gene transcription. When we investigated the
effect of IL-10 on the IFN-induced formation of ISGF3 (Fig 2A), we noted that there was a clear dose
response with respect to IFN concentrations, such that at 100 U/mL
(0.5 ng/mL) of IFN, inhibition by IL-10 was greater than 90%, yet
at 1,000 U/mL of IFN, inhibition by IL-10 had decreased to
approximately 35% (Fig 2B). This result suggested that formation of
ISGF3 was markedly inhibited by IL-10 at lower doses of IFN.
However, higher doses of IFN could overcome to some extent this
inhibition by increased receptor occupancy, suggesting that there may
be a limiting amount of an inhibitory component induced by IL-10 whose
function can be competed for when more IFN binds to the cell. The
addition of exogenous p48 to the reactions did not affect the ability
of the complex to bind to the ISRE probe (data not shown). When a similar experiment was performed using the GRR probe, which is recognized by STAT1 dimers, the same result was observed (Fig 2C).
Because binding to the GRR probe probably occurs through a mechanism
different from that to the ISRE (ie, neither p48 nor STAT2 is
involved), this result implies that STAT1 may potentially be a target
of IL-10 suppression.
View larger version (23K):
[in this window]
[in a new window]
| Fig 2.
EMSA analysis of IFN-induced DNA binding complexes.
Monocytes were treated with 10 ng/mL of IL-10 for 90 minutes, followed
by treatment with the indicated doses of IFN for 30 minutes. Nuclear
extracts were prepared, and EMSA were performed. (A) EMSA using the
ISRE probe, which represents binding of the ISGF3 complex. (B)
Summaries of the Phospho-Imager data from several experiments, such as
those presented in (A), were quantitated using the Phospho-Imager. The
value given is the mean ± standard deviation. (C) EMSA using the GRR
probe, which predominantly measures binding of STAT1 dimers.
|
|
We next examined the effect of IL-10 on IFN activation of STAT1
directly through the use of an antibody that recognizes only the
tyrosine-phosphorylated form of STAT1 (Fig
3A). IL-10 pretreatment of monocytes
resulted in reduced levels of tyrosine-phosphorylated STAT1 when cells
were treated with lower doses of IFN (200 U/mL or lower). This
result correlated with IL-10-mediated inhibition of IFN-induced gene
expression and formation of ISGF3. All lanes expressed equal amounts of
STAT1 loaded onto the gel (Fig 3B). With increasing doses of IFN,
there was a decrease in the inhibitory action of IL-10 on STAT1
tyrosine phosphorylation (Fig 3A, lanes 5 and 6), such that at 1,000 U/mL (5 ng/mL), inhibition was still present, but reduced to
approximately 40% (Fig 3C). This was consistent with the results
observed for gene induction and ISRE binding.
View larger version (20K):
[in this window]
[in a new window]
| Fig 3.
(A) IL-10 inhibits the IFN-induced tyrosine
phosphorylation of STAT1. Monocytes were treated with 10 ng/mL of IL-10
for 60 minutes, followed by treatment for 30 minutes with the indicated
doses of IFN at 200 U/mL (lanes 3 and 4) or 1,000 U/mL (lanes 5 and
6). Extracts were prepared, and equal amounts of protein were applied
to an SDS-PAGE gel. Separated proteins were transferred and a Western
blot was performed with an antibody that recognizes
tyrosine-phosphorylated STAT1. (B) The blot was stripped and reprobed
for STAT1. (C) The inhibitory effects of IL-10 on IFN-stimulated
phosphorylation of STAT1 are dose-dependent. Monocytes were prepared as
in (A), and the resulting autoradiographs were scanned for
phosphorylated STAT1 using a Laser densitometer (LKB-Brommo,
Piscataway, NJ). The numbers in parentheses refer to the
number of experiments performed with different donors. The data are
presented as the mean ± standard deviation.
|
|
Pretreatment of cells with IL-10 also suppressed the ability of
IFN-stimulated STAT1 dimeric complexes to bind to the GRR probe (Fig
4A, lane 3 v 4). There was no
evidence of IL-10-stimulated STAT binding to the GRR probe, because
the time of treatment with IL-10 (t = 90 minutes) before IFN was
sufficient for this binding to have dissipated. Under these conditions,
IL-10 inhibited IFN activation of GRR-binding complexes at several
concentrations of IFN (Fig 4B). The inability to repeatedly scan the
same exact region in EMSA autoradiographs from different experiments
leads to the variation observed in the plot. We also looked directly at
the ability of IL-10 to block activation of STAT1 using phosphotyrosine immunoblotting. Monocytes were pretreated with IL-10 and then incubated
with IFN for 15 minutes. The cells were lysed, and phosphorylation
of STAT1 was assessed with an antiphosphotyrosine antibody. There was
substantial inhibition of STAT1 tyrosine phosphorylation (Fig 5A, lanes
3 and 4). The levels of tyrosine
phosphorylation of both the 91-kD and a lower molecular weight STAT1
(Fig 5A, asterisk) were markedly decreased by IL-10 pretreatment (Fig
5A, lower panel). Because the preincubation of monocytes with vanadate, a phosphotyrosine phosphatase (PTP) inhibitor, had no effect on IL-10
inhibition (data not shown), it is unlikely that SHP1, a PTP, induces
the effects of IL-10. Moreover, because only tyrosine-phosphorylated STAT1 translocates to the nucleus, a decrease in tyrosine
phosphorylation implies an IL-10 event must occur at the receptor.
Again, we observed a dose-response effect of the inhibition, such that
at higher doses of IFN, the suppressive effects of IL-10 were much
less evident (Fig 5B). However, in most inflammatory conditions, IFN concentrations are generally low, and the inhibitory effect of IL-10 is
marked at concentrations of 1 ng/mL, in which activation of genes is
generally at a near maximal level.
View larger version (19K):
[in this window]
[in a new window]
| Fig 4.
Effect of IL-10 on IFN-induced assembly of STAT1
dimers. (A) Monocytes were preincubated with 10 ng/mL of IL-10 for 90 minutes and then incubated with IFN for 20 minutes. Whole-cell
extracts were prepared, and an EMSA was performed using the GRR probe,
which is recognized by phosphorylated STAT1. The upper band of the
shifted complex was presumed to be multimers of STAT1 binding to the
probe. The autoradiograph represents that part of the total gel
analyzed. (B) EMSA with GRR probe was performed as in (A), except that
the indicated doses of IFN were used. The response was measured as a
percentage of the maximum response of cells treated with IFN at 10 ng/mL. Three experiments were performed using individual donors, and
the data are presented as the mean ± standard error. ( ) Treated
with IFN alone; ( ) treated with IL-10 and IFN.
|
|
View larger version (19K):
[in this window]
[in a new window]
| Fig 5.
IL-10 inhibits the IFN-induced tyrosine
phosphorylation of STAT1. (A) Monocytes were pretreated with 10 ng/mL
of IL-10 for 60 minutes, and then treated with 0.5 ng/mL of IFN for
20 minutes. The upper panel represents an anti-STAT1
immunoprecipitation blotted with antiphosphotyrosine antibody. The
lower panel represents the same membrane reprobed with anti-STAT1. The
slower migrating band is tyrosine-phosphorylated STAT1
(STAT1P). The asterisk refers to a lower molecular weight
band that is frequently observed in the immunoprecipitations and is
related to STAT1 activation. (B) Monocytes were prepared as in (A),
except that the indicated doses of IFN were used. Summary of laser
densitometric scans of multiple antiphosphotyrosine immunoblots is
shown. The values for percentage of inhibition were calculated, and the
data are presented as the mean ± standard deviation for the
indicated number of independent experiments.
|
|
Recently, a series of genes have been cloned that are known to inhibit
cytokine-induced activation of cells.36-39 To determine if
IL-10 induced the expression of any of these genes, we treated cells
with IL-10 and isolated RNA for Northern blot analysis. The results of
this analysis showed that IL-10 could rapidly (within 30 minutes)
induce the expression of the SOCS3 gene and maintain its expression for
at least 120 minutes after the addition of IL-10 (Fig
6). The kinetics of this
induction of SOCS3 gene expression by IL-10 were consistent with its
potential role in counterregulating IFN-induced STAT1 activation and
IFN-induced gene expression. In preliminary experiments, we found that
IL-10 did not upregulate expression of the related genes, CIS-1,
SOCS-1, or SOCS-2, in monocytes (data not shown).
View larger version (64K):
[in this window]
[in a new window]
| Fig 6.
IL-10 induces expression of the SOCS3 gene in monocytes.
Monocytes were incubated for the designated times with 10 ng/mL of
IL-10 and then processed for RNA isolation. Northern blot analysis was
performed as outlined in the Materials and Methods.
|
|
| |
DISCUSSION |
IL-10 suppresses the ability of LPS to elicit increases in mRNA of
IL-1, tumor necrosis factor, IL-6, and other cytokines or cytokine
receptors.13 In some of these studies, it has also been
shown that the ability of IFN to enhance the effects of LPS can also
be suppressed by IL-10.40 However, there has been no study
addressing whether IL-10 inhibits IFN- and IFN-stimulated early
response genes in monocytes. Both IFN and IFN induce the expression of these genes by activating STAT proteins that bind to
either GAS or ISRE enhancers.16 In this report, we showed that IL-10 can inhibit the induction of IFN-induced genes by preventing the assembly of ISGF3 and GRR/GAS transcription complexes. The suppression of gene expression correlates with the IL-10-induced inhibition of the tyrosine phosphorylation of STAT1, which is required
for this protein to bind DNA. Moreover, we have shown that the SOCS3
gene is induced by IL-10 and may be involved in the suppression of
STAT1 phosphorylation.
IL-10 has numerous effects on hematopoietic cells, but most notable is
its association with the development of Th2 lymphocytes. The
polarization of T-cell development is thought to progress from a Th0
phase to either a Th1 or Th2 cell, depending on the presence or absence
of various ILs.41 IL-10 is secreted by Th2 cells, and once
secreted, it plays a major role in inhibition of monocyte/macrophage
function, thereby suppressing the Th1 phenotype.42 Like
IFNs, IL-10 activates STAT complexes.15,17,43,44 Incubation of monocytes with IL-10 activates primarily STAT3 and, to a lesser extent, STAT1.15 However, studies using STAT1-deficient
mice indicated that STAT3 is important for IL-10 inhibition of
LPS-induced monocyte activation.28 Because IL-10 activation
of STATs is transient in monocytes (<30 minutes), by the time IFNs
were added, activated STATs were not present (data not shown).
Because IL-10 treatment of monocytes does not alter the number or
affinity of IFN cell surface receptors, another mechanism must be
regulated by which IL-10 inhibits IFN-stimulated phosphorylation of
STAT1. There has been recently discovered a new class of molecules referred to as SOCS proteins that seem to be induced by cytokines and
then act as negative regulators to inhibit signal
transduction.36-39 These molecules possess SH2 domains and
can bind to phosphotyrosines on Jaks or cytokine receptors. Of
interest, Jak1 is used by all three molecules: IFN, IFN, and
IL-10. There does seem to be some specificity with regard to the
inhibitory actions of SOCS, because only SOCS1 and 3, but not CIS (an
inhibitor of GM-CSF-induced STAT5), were able to inhibit growth
hormone-induced STAT5 activation.45 Constitutive SOCS1
expression in transfected cells was shown to suppress IFN
activation.36 No studies have shown whether IL-10 induces
the expression of any SOCS genes. Experiments to detect association
between Jaks and SOCS proteins have been very difficult using available
antibodies. Only very high expression of a transiently transfected
myc-JAB (CIS-like) gene and Jak2 (that is constitutively tyrosine
phosphorylated) allowed coimmunoprecipitation of these two
proteins.38 In this report, we do show that IL-10 can
rapidly (30 minutes) induce the expression of SOCS3, possibly through its ability to induce tyrosine phosphorylation of STAT3. Whether STAT3-stimulated expression of SOCS3 accounts for the IL-10 inhibition of STAT1 tyrosine phosphorylation by IFNs is not yet known.
Another well-documented mechanism to inhibit cytokine signaling is the
activation of tyrosine phosphatases that inactivate Jaks by
dephosphorylation. Ligand-stimulated tyrosine phosphorylation of the
erythropoietin receptor can be inhibited by targeting the phosphatase
SHP1 (PTP-1C) to the receptor, resulting in dephosphorylation of
Jak2.46,47 Although we have found that the amount of Jak1 is unchanged after IL-10 treatment, under the current culture conditions we have not been able to quantitate its degree of tyrosine phosphorylation, either by in vitro kinase assay or by immunoblotting with antiphosphotyrosine. When the cells were incubated with vanadate, a tyrosine phosphatase inhibitor, there was no evidence that the inhibitory actions of IL-10 were altered, suggesting that
IL-10-regulated phosphatase activity is not involved (data not shown).
One common theme throughout the study has been the effect of the dose
of IFN on the inhibitory action of IL-10. At high doses of either
IFN or IFN, the inhibitory action of IL-10 is reduced. At lower
doses of IFN, less STAT1 is tyrosine phosphorylated in the presence of
IL-10, suggesting the possibility that an event may be occurring at the
plasma membrane that is regulated by IL-10. We and others have
previously shown that Jaks are associated with IFN receptors, and STAT1
migrates to the receptor once IFN binds. Because the major event that
determines the amount of tyrosine phosphorylation of STAT1 is the
concentration of IFN binding to its receptor, it seems reasonable to
hypothesize that the effect of IL-10 is to disrupt this multimolecular
complex, such that the efficiency of STAT1 tyrosine phosphorylation is
diminished. IL-10 induction of SOCS3 would be one such mechanism to
account for the observed inhibitory actions of IL-10.
We have shown that IL-10 can inhibit the induction of certain
IFN-induced genes. ICAM-1 was only induced by IFN and showed good
IL-10-induced inhibition. The inhibition by IL-10 was fairly consistent among the genes, except for IRF-1. Although IRF-1 has a GAS
motif that drives this gene, there was only a minimal effect by
IL-10.48 This gene was readily induced by IFN in
monocytes, but no induction was observed with IFN (data not shown).
There are several other elements in the promoter of this gene that may override IFN activation through its GAS element, if this enhancer is
inhibited by incubation of cells with IL-10. These include Sp1 and
NFB binding sites. Recently, mice deficient in the IFN-induced-gene, double-stranded RNA-activated Ser/Thr protein kinase (PKR) were studied
for their responsiveness to IFN.49 PKR is important for
IFN-induced activation of NFB. Cells from these mice showed no
IFN-induced expression of IRF-1-promoter-reporter constructs. This
response was restored on transfection with wild-type PKR, suggesting
that NFB may play an important role in mediating the expression of
IRF-1.
It was recently shown that in human monocytes, there was no inhibition
of GAS binding activity or STAT1 tyrosine phosphorylation after
pretreatment with IL-10 in a study of ICAM-1 induction by IFN.34 One possible reason for the discrepancy between
our results is the differences in the way the monocytes were prepared. In Song et al,34 the monocytes were passed over
Ficoll-Hypaque and then allowed to adhere for 1.5 hours in medium
containing 10% fetal bovine serum. This process allows for adherence
of some B cells and possibly even some T cells. These monocyte
preparations were generally only 85% pure. Moreover, Song et
al34 used a concentration of IFN (10 ng/mL) that was
10-fold higher than the maximal concentrations of IFN we used to
observe the effects described in this report.27 We observed
a decrease in IFN-induced STAT binding to two distinct elements and a
decrease in the corresponding STAT1 tyrosine phosphorylation for both
IFN and IFN. This suggests that, for cells isolated by our method
with minimal handling and more physiological concentrations of IFN,
incubation with IL-10 inhibits IFN-induced gene activation by
preventing the phosphorylation of STAT1 that may occur through
activation of a new set of genes that encode for proteins that suppress
continued cytokine activation.24 The availability of
antibodies to these molecules will enable further characterization of
their mechanism of action. Our findings define an important mechanism
by which IL-10 can antagonize IFN-induced gene expression in monocytes.
| |
ACKNOWLEDGMENT |
The authors thank Dr D. Hilton (Walter and Eliza Hall Institute,
Parkville, Australia) for providing the SOCS cDNAs; Genentech, Inc
(South San Francisco, CA) for IFN; Roche, Inc (Nutley, NJ) for
IFN; and Schering-Plough, Inc (Kenilworth, NJ) for IL-10.
| |
FOOTNOTES |
Submitted May 11, 1998; accepted October 12, 1998.
Supported by the Oak Ridge Institute for Science and Education through
an interagency agreement between the Department of Energy and the FDA
(S.I., H.D., and N.V.).
The publication costs of this
article were defrayed in part by
page charge payment. This article
must therefore be hereby marked
"advertisement"
in accordance with 18 U.S.C. section
1734 solely to indicate this fact.
Address reprint requests to David S. Finbloom, MD, Division of Cytokine
Biology, Center for Biologics Evaluation and Research, Food and Drug
Administration, 29 Lincoln Dr (Bldg 29A, Room 2D20), Bethesda, MD
20892-4555.
| |
REFERENCES |
1.
Hagenbeek A, Carde P, Meerwaldt JH, Somers R, Thomas J, De Bock R, Raemaekers JM, van Hoof A, De W:
Human recombinant interferon alfa-2a in patients with stages III and IV low-grade malignant non-Hodgkin's lymphoma. European Organization for Research and Treatment of Cancer Lymphoma Cooperative Group.
J Clin Oncol
16:41, 1998[Abstract/Free Full Text]
2.
Zinzani PL, Bendandi M, Magagnoli M, Rondelli D, de Vivo A, Benni M, Zamagni E, Cavo M, Tura S:
Results of a fludarabine induction and alpha-interferon maintenance protocol in pretreated patients with chronic lymphocytic leukemia and low-grade non-Hodgkin's lymphoma.
Eur J Haematol
59:82, 1997[Medline]
[Order article via Infotrieve]
3.
Tamm I, Grimme H, Bergen E, Simon JC, Schopf E, Mertelsmann R, Lindemann A, Brennscheidt U:
Dacarbazine and interferon alpha for stage IV malignant melanoma.
Oncology
54:270, 1997[Medline]
[Order article via Infotrieve]
4.
Biron C, Gazzinelli R:
Effects of IL-12 on immune responses to microbial infections: A key mediator in regulating disease outcome.
Curr Opin Immunol
7:485, 1995[Medline]
[Order article via Infotrieve]
5.
Petricoin E, David M, Fang H, Grimley P, Larner AC, Vande Pol S:
Human cancer cell lines express a negative transcriptional regulator of the interferon regulatory factor family of DNA binding proteins.
Mol Cell Biol
14:1477, 1994[Abstract/Free Full Text]
6.
Finbloom DS, Larner AC, Nakagawa Y, Hoover DL:
Culture of human monocytes in granulocyte-macrophage CSF results in enhancement of interferon receptors but suppression of IFN-induced expression of the gene, IP-10.
J Immunol
150:2383, 1993[Abstract]
7.
O'Garra A, Murphy K:
Role of cytokines in determining T-lymphocyte function.
Curr Opin Immunol
6:458, 1994[Medline]
[Order article via Infotrieve]
8.
Locksley RM, Scott P:
Helper T cell subsets in mouse leishmaniasis: Induction, expansion and effector function.
Immunol Today
12:58, 1991
9.
Katsikis PD, Chu CQ, Brennan FM, Maini RN, Feldmann M:
Immunoregulatory role of interleukin 10 in rheumatoid arthritis.
J Exp Med
179:1517, 1994[Abstract/Free Full Text]
10.
Powrie F, Leach MW, Mauze S, Monon S, Caddle LB, Coffman RL:
Inhibition of Th1 responses prevents inflammatory bowel disease in scid mice reconstituted with CD45RBhi CD4+ T cells.
Immunity
1:553, 1994[Medline]
[Order article via Infotrieve]
11.
Larner AC, Petricoin EF, Nakagawa Y, Finbloom DS:
IL-4 attenuates the transcriptional activation of both IFN and IFN induced cellular gene expression in monocytes and monocytic cell lines.
J Immunol
150:1944, 1993[Abstract]
12.
Dickensheets HL, Donnelly RP:
IFN- and IL-10 inhibit induction of IL-1 receptor type-1 and type-II gene expression by IL-4 and IL-13 in human monocytes.
J Immunol
159:6226, 1997[Abstract]
13.
De Waal Malefyt R, Abrans J, Bennett B, Figdor C, de Vries J:
IL-10 inhibits cytokine synthesis by human monocytes: An autoregulatory role of IL-10 produced by monocytes.
J Exp Med
174:1209, 1991[Abstract/Free Full Text]
14.
Finbloom DS, Larner AC:
Induction of early response genes by interferons, interleukins, and growth factors by the tyrosine phosphorylation of latent transcription factors: Implications for inflammatory diseases.
Arthritis Rheum
38:877, 1995[Medline]
[Order article via Infotrieve]
15.
Finbloom DS, Winestock KD:
IL-10 induces the tyrosine phosphorylation of tyk2 and Jak1 and the differential assembly of STAT1a and STAT3 complexes in human T cells and monocytes.
J Immunol
155:1079, 1995[Abstract]
16.
Larner AC, Finbloom DS:
Protein tyrosine phosphorylation as a mechanism which regulates cytokine activation of early response genes.
Biochim Biophys Acta
1266:278, 1995[Medline]
[Order article via Infotrieve]
17.
Larner AC, David M, Feldman GM, Igarashi K, Hackett RH, Webb DAS, Sweitzer SM, Petricoin EF III, Finbloom DS:
Tyrosine phosphorylation of DNA binding proteins by multiple cytokines.
Science
261:1730, 1993[Abstract/Free Full Text]
18.
Akai H, Larner AC:
Phorbol ester-mediated down-regulation of an interferon-inducible gene.
J Biol Chem
264:3252, 1989[Abstract/Free Full Text]
19.
Luster AD, Unkeless JC, Ravetch JV:
-Interferon transcriptionally regulates an early-response gene containing homology to platelet proteins.
Nature
315:672, 1985[Medline]
[Order article via Infotrieve]
20.
Akai H, Larner AC:
Characterization of two independent mechanisms by which interferon-induced gene expression is down regulated.
Cell Regul
1:707, 1990[Medline]
[Order article via Infotrieve]
21.
Cheng YE, Patterson CE, Staeheli P:
Interferon-induced guanylate-binding proteins lack an N(T)KXD concensus motif and bind GMP in addition to GDP AND GTP.
Mol Cell Biol
11:4717, 1991[Abstract/Free Full Text]
22.
Piechaczyk M, Blanchard JM, Marty L, Dani C, Panabieres F, El Sabouty S, Fort P, Jeanteur P:
Post-transcriptional regulation of glyceraldehyde-3-phosphate-dehydrogenase gene expression in rat tissues.
Nucleic Acids Res
12:6951, 1984[Abstract/Free Full Text]
23.
Chomczynski P, Sacchi N:
Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction.
Anal Biochem
162:156, 1987[Medline]
[Order article via Infotrieve]
24.
Hilton DJ, Richardson RT, Alexander WS, Viney EM, Willson TA, Sprigg NS, Starr R, Nicholson SE, Metcalf D, Nicola NA:
Twenty proteins containing a C-terminal SOCS box form five structural classes.
Proc Natl Acad Sci USA
95:114, 1998[Abstract/Free Full Text]
25.
Feinberg AP, Vogelstein B:
A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity.
Anal Biochem
132:6, 1983[Medline]
[Order article via Infotrieve]
26.
Pearse RN, Feinman R, Ravetch JV:
Characterization of the promoter of the human gene encoding the high affinity IgG receptor: Transcriptional induction by -interferon is mediated through common DNA response elements.
Proc Natl Acad Sci USA
88:11305, 1991[Abstract/Free Full Text]
27.
Wilson KC, Finbloom DS:
Interferon rapidly induces in human monocytes a DNA-binding factor that recognizes the response region within the promoter of the gene for the high-affinity Fc receptor.
Proc Natl Acad Sci USA
89:11964, 1992[Abstract/Free Full Text]
28.
Meraz MA, White JM, Sheehan KC, Bach EA, Rodig SJ, Dighe AS, Kaplan DH, Riley JK, Greenlund AC, Campbell D, Carver-Moore K, DuBois RN, Clark R, Aguet M, Schreiber RD:
Targeted disruption of the Stat1 gene in mice reveals unexpected physiologic specificity in the JAK-STAT signaling pathway.
Cell
84:431, 1996[Medline]
[Order article via Infotrieve]
29.
Luster AD, Greenberg SM, Leder P:
The IP-10 chemokine binds to a specific cell surface heparin sulfate site shared with platelet factor 4 and inhibits endothelial cell proliferation.
J Exp Med
182:219, 1995[Abstract/Free Full Text]
30.
Sgadari C, Angiolillo AL, Cherney BW, Pike SE, Farber JM, Koniaris LG, Vanguri P, Burd PR, Sheikh N, Gupta G, Teruya-Feldstein J, Tosato G:
Interferon-inducible protein-10 identified as a mediator of tumor necrosis in vivo.
Proc Natl Acad Sci USA
93:13791, 1996[Abstract/Free Full Text]
31.
Xia Y, Pauza ME, Feng L, Lo D:
RelB regulation of chemokine expression modulates local inflammation.
Am J Pathol
151:375, 1997[Abstract]
32.
Terol MJ, Lopez-Guillermo A, Bosch F, Villamor N, Cid MC, Rozman C, Campo E, Montserrat E:
Expression of the adhesyion molecule ICAM-1 in non-Hodgkin's lymphoma: Relationship with tumor dissemination and prognostic importance.
J Clin Oncol
16:35, 1998[Abstract/Free Full Text]
33.
Croft M, Dubey C:
Accessory molecule and costimulation requirements for CD4 T cell response.
Crit Rev Immunol
17:89, 1997[Medline]
[Order article via Infotrieve]
34.
Song S, Ling-Hu H, Roebuck KA, Rabbi MF, Donnelly RP, Finnegan A:
Interleukin-10 inhibits interferon-gamma-induced intercellular adhesion molecule-1 gene transcription in human monocytes.
Blood
89:4461, 1997[Abstract/Free Full Text]
35.
Eilers A, Seegert D, Schindler C, Baccarini M, Decker T:
The response of gamma interferon activation factor is under developmental control in cells of the macrophage lineage.
Mol Cell Biol
13:3245, 1993[Abstract/Free Full Text]
36.
Starr R, Willson TA, Viney EM, Murray LJL, Rayner RJR, Jenkin BJ, Gonda TJ, Alexander WS, Metcalf D, Nicola NA, Hilton DJ:
A family of cytokine-inducible inhibitors of signalling.
Nature
387:917, 1997[Medline]
[Order article via Infotrieve]
37.
Naka T, Narazaki M, Hirata M, Matsumoto T, Minamoto S, Aono A, Nishimoto N, Kajita T, Taga T, Yoshizaki K, Akira S, Kishimoto T:
Structure and function of a new STAT-induced STAT inhibitor.
Nature
387:924, 1997[Medline]
[Order article via Infotrieve]
38.
Endo TA, Masuhara M, Yokouchi M, Suzuki R, Sakamoto H, Mitsui K, Matsumoto A, Tanimura S, Ohtsubo M, Misawa H, Miyazaki T, Leonor N, Taniguchi T, Fujita T, Kanakura Y, Komiya S, Yoshimura A:
A new protein containing an SH2 domain that inhibits JAK kinases.
Nature
387:921, 1997[Medline]
[Order article via Infotrieve]
39.
Matsumoto A, Masuhara M, Mitsui K, Yokouchi M, Ohtsubo M, Misawa H, Miyajima A, Yoshimura A:
CIS, a cytokine inducible SH2 protein, is a target of the JAK-STAT5 pathway and modulates STAT5 activation.
Blood
89:3148, 1997[Abstract/Free Full Text]
40.
Gazzinelli RT, Oswald IP, James SL, Sher A:
IL-10 inhibits parasite killing and nitric oxide production by IFN--activated macrophages.
J Immunol
148:1792, 1992[Abstract]
41.
O'Garra A, Murphy K:
Role of cytokines in determining T-lymphocyte function.
Curr Opin Immunol
6:458, 1994
42.
Moore KW, O'Garra A, de Waal Malefyt R, Vieira P, Mosmann TR:
Interleukin-10.
Annu Rev Immunol
11:165, 1993[Medline]
[Order article via Infotrieve]
43.
Lehmann J, Seegert D, Strehlow I, Schindler C, Lohmann-Matthes M-L, Decker T:
IL-10-induced factors belonging to the p91 family of proteins bind to IFN--responsive promoter elements.
J Immunol
153:165, 1994[Abstract]
44.
Wehinger J, Gouilleux G, Groner B, Finke J, Mertelsmann R, Weber-Nordt RM:
IL-10 induces DNA binding activity of three STAT proteins (Stat1, Stat3, and Stat5) and their distinct combinatorial assembly in the promoters of selected genes.
FEBS Lett
394:365, 1996[Medline]
[Order article via Infotrieve]
45.
Adams TE, Hansen JA, Starr R, Nicola NA, Hilton DJ, Billestrup N:
Growth hormone preferentially induces the rapid, transient expression of SOCS-3, a novel inhibitor of cytokine receptor signalling.
J Biol Chem
273:1285, 1998[Abstract/Free Full Text]
46.
Klingmuller U, Lorenz U, Cantley LC, Neel BG, Lodish HF:
Specific recruitment of SH-PTP1 to the erythropoietin receptor causes inactivation of Jak2 and termination of proliferative signals.
Cell
80:729, 1995[Medline]
[Order article via Infotrieve]
47.
Pawson T:
Protein modules and signalling networks.
Nature
373:573, 1995[Medline]
[Order article via Infotrieve]
48.
Sims SH, Cha Y, Romine MF, Gao P-Q, Gottlieb K, Deisseroth AB:
A novel interferon-inducible domain: Structural and functional analysis of the human interferon regulatory factor 1 gene promoter.
Mol Cell Biol
13:690, 1993[Abstract/Free Full Text]
49.
Kumar A, Yang YL, Flati V, Der S, Kadereit S, Deb A, Haque J, Reis L, Weissmann C, Williams BR:
Deficient cytokine signaling in mouse embryo fibroblasts with a targeted deletion in the PKR gene: Role of IRF-1 and NF-kappaB.
EMBO J
15:406, 1997
This is a US government work. There are no restrictions on its use.
0006-4971/99/9305-0120$0.00/0
 CiteULike  Connotea  Del.icio.us  Digg  Reddit  Technorati What's this?
This article has been cited by other articles:
|
|
|
|
 
M. Sarasin-Filipowicz, X. Wang, M. Yan, F. H. T. Duong, V. Poli, D. J. Hilton, D.-E. Zhang, and M. H. Heim
Alpha Interferon Induces Long-Lasting Refractoriness of JAK-STAT Signaling in the Mouse Liver through Induction of USP18/UBP43
Mol. Cell. Biol.,
September 1, 2009;
29(17):
4841 - 4851.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. S. Bharhani, B. Chiu, K.-S. Na, and R. D. Inman
Activation of invariant NKT cells confers protection against Chlamydia trachomatis-induced arthritis
Int. Immunol.,
July 1, 2009;
21(7):
859 - 870.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
R. J. Critchley-Thorne, D. L. Simons, N. Yan, A. K. Miyahira, F. M. Dirbas, D. L. Johnson, S. M. Swetter, R. W. Carlson, G. A. Fisher, A. Koong, et al.
Impaired interferon signaling is a common immune defect in human cancer
PNAS,
June 2, 2009;
106(22):
9010 - 9015.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
D. Miyazaki, C. H. Kuo, T. Tominaga, Y. Inoue, and S. J. Ono
Regulatory Function of CpG-Activated B Cells in Late-Phase Experimental Allergic Conjunctivitis
Invest. Ophthalmol. Vis. Sci.,
April 1, 2009;
50(4):
1626 - 1635.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
W.-S. Lim, J. M. Timmins, T. A. Seimon, A. Sadler, F. D. Kolodgie, R. Virmani, and I. Tabas
Signal Transducer and Activator of Transcription-1 Is Critical for Apoptosis in Macrophages Subjected to Endoplasmic Reticulum Stress In Vitro and in Advanced Atherosclerotic Lesions In Vivo
Circulation,
February 19, 2008;
117(7):
940 - 951.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
D. Holzinger, C. Jorns, S. Stertz, S. Boisson-Dupuis, R. Thimme, M. Weidmann, J.-L. Casanova, O. Haller, and G. Kochs
Induction of MxA Gene Expression by Influenza A Virus Requires Type I or Type III Interferon Signaling
J. Virol.,
July 15, 2007;
81(14):
7776 - 7785.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
X. Zhang, E. Deriaud, X. Jiao, D. Braun, C. Leclerc, and R. Lo-Man
Type I interferons protect neonates from acute inflammation through interleukin 10-producing B cells
J. Exp. Med.,
May 14, 2007;
204(5):
1107 - 1118.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. S. Bharhani, R. Borojevic, S. Basak, E. Ho, P. Zhou, and K. Croitoru
IL-10 protects mouse intestinal epithelial cells from Fas-induced apoptosis via modulating Fas expression and altering caspase-8 and FLIP expression.
Am J Physiol Gastrointest Liver Physiol,
November 1, 2006;
291(5):
G820 - G829.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
N. Vazquez, T. Greenwell-Wild, S. Rekka, J. M. Orenstein, and S. M. Wahl
Mycobacterium avium-induced SOCS contributes to resistance to IFN-{gamma}-mediated mycobactericidal activity in human macrophages
J. Leukoc. Biol.,
November 1, 2006;
80(5):
1136 - 1144.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
V. A. Dennis, A. Jefferson, S. R. Singh, F. Ganapamo, and M. T. Philipp
Interleukin-10 Anti-Inflammatory Response to Borrelia burgdorferi, the Agent of Lyme Disease: a Possible Role for Suppressors of Cytokine Signaling 1 and 3.
Infect. Immun.,
October 1, 2006;
74(10):
5780 - 5789.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
E. Giacomini, A. Sotolongo, E. Iona, M. Severa, M. E. Remoli, V. Gafa, R. Lande, L. Fattorini, I. Smith, R. Manganelli, et al.
Infection of Human Dendritic Cells with a Mycobacterium tuberculosis sigE Mutant Stimulates Production of High Levels of Interleukin-10 but Low Levels of CXCL10: Impact on the T-Cell Response.
Infect. Immun.,
June 1, 2006;
74(6):
3296 - 3304.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
P. Qasimi, A. Ming-Lum, A. Ghanipour, C. J. Ong, M. E. Cox, J. Ihle, N. Cacalano, A. Yoshimura, and A. L-F. Mui
Divergent Mechanisms Utilized by SOCS3 to Mediate Interleukin-10 Inhibition of Tumor Necrosis Factor {alpha} and Nitric Oxide Production by Macrophages
J. Biol. Chem.,
March 10, 2006;
281(10):
6316 - 6324.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
B. M. Sullivan, O. Jobe, V. Lazarevic, K. Vasquez, R. Bronson, L. H. Glimcher, and I. Kramnik
Increased Susceptibility of Mice Lacking T-bet to Infection with Mycobacterium tuberculosis Correlates with Increased IL-10 and Decreased IFN-{gamma} Production
J. Immunol.,
October 1, 2005;
175(7):
4593 - 4602.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J. S. Satoguina, E. Weyand, J. Larbi, and A. Hoerauf
T Regulatory-1 Cells Induce IgG4 Production by B Cells: Role of IL-10
J. Immunol.,
April 15, 2005;
174(8):
4718 - 4726.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
H. Ooboshi, S. Ibayashi, T. Shichita, Y. Kumai, J. Takada, T. Ago, S. Arakawa, H. Sugimori, M. Kamouchi, T. Kitazono, et al.
Postischemic Gene Transfer of Interleukin-10 Protects Against Both Focal and Global Brain Ischemia
Circulation,
February 22, 2005;
111(7):
913 - 919.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
D. Spight, B. Zhao, M. Haas, S. Wert, A. Denenberg, and T. P. Shanley
Immunoregulatory effects of regulated, lung-targeted expression of IL-10 in vivo
Am J Physiol Lung Cell Mol Physiol,
February 1, 2005;
288(2):
L251 - L265.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. Prabhakar, Y. Qiao, A. Canova, D. B. Tse, and R. Pine
IFN-{alpha}{beta} Secreted during Infection Is Necessary but Not Sufficient for Negative Feedback Regulation of IFN-{alpha}{beta} Signaling by Mycobacterium tuberculosis
J. Immunol.,
January 15, 2005;
174(2):
1003 - 1012.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
G. Grutz
New insights into the molecular mechanism of interleukin-10-mediated immunosuppression
J. Leukoc. Biol.,
January 1, 2005;
77(1):
3 - 15.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
V. S. Carl, J. K. Gautam, L. D. Comeau, and M. F. Smith Jr
Role of endogenous IL-10 in LPS-induced STAT3 activation and IL-1 receptor antagonist gene expression
J. Leukoc. Biol.,
September 1, 2004;
76(3):
735 - 742.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
R. P. Donnelly, F. Sheikh, S. V. Kotenko, and H. Dickensheets
The expanded family of class II cytokines that share the IL-10 receptor-2 (IL-10R2) chain
J. Leukoc. Biol.,
August 1, 2004;
76(2):
314 - 321.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
D. E. Spaner
Amplifying cancer vaccine responses by modifying pathogenic gene programs in tumor cells
J. Leukoc. Biol.,
August 1, 2004;
76(2):
338 - 351.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. Mazighi, A. Pelle, W. Gonzalez, E. M. Mtairag, M. Philippe, D. Henin, J.-B. Michel, and L. J. Feldman
IL-10 inhibits vascular smooth muscle cell activation in vitro and in vivo
Am J Physiol Heart Circ Physiol,
August 1, 2004;
287(2):
H866 - H871.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
C. Carbonneil, V. Donkova-Petrini, A. Aouba, and L. Weiss
Defective Dendritic Cell Function in HIV-Infected Patients Receiving Effective Highly Active Antiretroviral Therapy: Neutralization of IL-10 Production and Depletion of CD4+CD25+ T Cells Restore High Levels of HIV-Specific CD4+ T Cell Responses Induced by Dendritic Cells Generated in the Presence of IFN-{alpha}
J. Immunol.,
June 15, 2004;
172(12):
7832 - 7840.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
P. Perrier, F. O. Martinez, M. Locati, G. Bianchi, M. Nebuloni, G. Vago, F. Bazzoni, S. Sozzani, P. Allavena, and A. Mantovani
Distinct Transcriptional Programs Activated by Interleukin-10 with or without Lipopolysaccharide in Dendritic Cells: Induction of the B Cell-Activating Chemokine, CXC Chemokine Ligand 13
J. Immunol.,
June 1, 2004;
172(11):
7031 - 7042.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
P. H. Correll, A. C. Morrison, and M. A. Lutz
Receptor tyrosine kinases and the regulation of macrophage activation
J. Leukoc. Biol.,
May 1, 2004;
75(5):
731 - 737.
[Full Text]
[PDF]
|
|
|
|
|
|
|
H.-J. Kim, J. Hart, N. Knatz, M. W. Hall, and M. D. Wewers
Janus Kinase 3 Down-Regulates Lipopolysaccharide-Induced IL-1{beta}-Converting Enzyme Activation by Autocrine IL-10
J. Immunol.,
April 15, 2004;
172(8):
4948 - 4955.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
W. Xiong, Y. Zhao, A. Prall, T. C. Greiner, and B. T. Baxter
Key Roles of CD4+ T Cells and IFN-{gamma} in the Development of Abdominal Aortic Aneurysms in a Murine Model
J. Immunol.,
February 15, 2004;
172(4):
2607 - 2612.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. C. Morrison, C. B. Wilson, M. Ray, and P. H. Correll
Macrophage-Stimulating Protein, the Ligand for the Stem Cell-Derived Tyrosine Kinase/RON Receptor Tyrosine Kinase, Inhibits IL-12 Production by Primary Peritoneal Macrophages Stimulated with IFN-{gamma} and Lipopolysaccharide
J. Immunol.,
February 1, 2004;
172(3):
1825 - 1832.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
O. Murillo, A. Arina, I. Tirapu, C. Alfaro, G. Mazzolini, B. Palencia, A. L.-D. De Cerio, J. Prieto, M. Bendandi, and I. Melero
Potentiation of Therapeutic Immune Responses against Malignancies with Monoclonal Antibodies
Clin. Cancer Res.,
November 15, 2003;
9(15):
5454 - 5464.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
K. Asadullah, W. Sterry, and H. D. Volk
Interleukin-10 Therapy--Review of a New Approach
Pharmacol. Rev.,
June 1, 2003;
55(2):
241 - 269.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J.-P. Herbeuval, C. Lambert, O. Sabido, M. Cottier, P. Fournel, M. Dy, and C. Genin
Macrophages From Cancer Patients: Analysis of TRAIL, TRAIL Receptors, and Colon Tumor Cell Apoptosis
J Natl Cancer Inst,
April 16, 2003;
95(8):
611 - 621.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
R. Condos, B. Raju, A. Canova, B.-Y. Zhao, M. Weiden, W. N. Rom, and R. Pine
Recombinant Gamma Interferon Stimulates Signal Transduction and Gene Expression in Alveolar Macrophages In Vitro and in Tuberculosis Patients
Infect. Immun.,
April 1, 2003;
71(4):
2058 - 2064.
[Abstract]
[Full Text]
|
|
|
|
|
|
|
H. Chen, L. Hutt-Fletcher, L. Cao, and S. D. Hayward
A Positive Autoregulatory Loop of LMP1 Expression and STAT Activation in Epithelial Cells Latently Infected with Epstein-Barr Virus
J. Virol.,
April 1, 2003;
77(7):
4139 - 4148.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. W. Fox, S. J. Haque, A. C. Lovibond, and T. J. Chambers
The Possible Role of TGF-{beta}-Induced Suppressors of Cytokine Signaling Expression in Osteoclast/Macrophage Lineage Commitment In Vitro
J. Immunol.,
April 1, 2003;
170(7):
3679 - 3687.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
C. Niemand, A. Nimmesgern, S. Haan, P. Fischer, F. Schaper, R. Rossaint, P. C. Heinrich, and G. Muller-Newen
Activation of STAT3 by IL-6 and IL-10 in Primary Human Macrophages Is Differentially Modulated by Suppressor of Cytokine Signaling 3
J. Immunol.,
March 15, 2003;
170(6):
3263 - 3272.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. Matsumoto, Y.-i. Seki, R. Watanabe, K. Hayashi, J. A. Johnston, Y. Harada, R. Abe, A. Yoshimura, and M. Kubo
A Role of Suppressor of Cytokine Signaling 3 (SOCS3/CIS3/SSI3) in CD28-mediated Interleukin 2 Production
J. Exp. Med.,
February 17, 2003;
197(4):
425 - 436.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
Y. Ding, D. Chen, A. Tarcsafalvi, R. Su, L. Qin, and J. S. Bromberg
Suppressor of Cytokine Signaling 1 Inhibits IL-10-Mediated Immune Responses
J. Immunol.,
February 1, 2003;
170(3):
1383 - 1391.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A.-F. Petit-Bertron, C. Fitting, J.-M. Cavaillon, and M. Adib-Conquy
Adherence influences monocyte responsiveness to interleukin-10
J. Leukoc. Biol.,
January 1, 2003;
73(1):
145 - 154.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
T. Ikeda, K. Sato, N. Kuwada, T. Matsumura, T. Yamashita, F. Kimura, K. Hatake, K. Ikeda, and K. Motoyoshi
Interleukin-10 differently regulates monocyte chemoattractant protein-1 gene expression depending on the environment in a human monoblastic cell line, UG3
J. Leukoc. Biol.,
December 1, 2002;
72(6):
1198 - 1205.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
T. Fumeaux and J. Pugin
Role of Interleukin-10 in the Intracellular Sequestration of Human Leukocyte Antigen-DR in Monocytes during Septic Shock
Am. J. Respir. Crit. Care Med.,
December 1, 2002;
166(11):
1475 - 1482.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. Mahalingam and B. A. Lidbury
Suppression of lipopolysaccharide-induced antiviral transcription factor (STAT-1 and NF-kappa B) complexes by antibody-dependent enhancement of macrophage infection by Ross River virus
PNAS,
October 15, 2002;
99(21):
13819 - 13824.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
I. Sakai, K. Takeuchi, H. Yamauchi, H. Narumi, and S. Fujita
Constitutive expression of SOCS3 confers resistance to IFN-alpha in chronic myelogenous leukemia cells
Blood,
September 26, 2002;
100(8):
2926 - 2931.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S Schreiber, P Rosenstiel, J Hampe, S Nikolaus, B Groessner, A Schottelius, T Kuhbacher, J Hamling, U R Folsch, and D Seegert
Activation of signal transducer and activator of transcription (STAT) 1 in human chronic inflammatory bowel disease
Gut,
September 1, 2002;
51(3):
379 - 385.
[Abstract]
[Full Text]
|
|
|
|
|
|
|
R. Lang, D. Patel, J. J. Morris, R. L. Rutschman, and P. J. Murray
Shaping Gene Expression in Activated and Resting Primary Macrophages by IL-10
J. Immunol.,
September 1, 2002;
169(5):
2253 - 2263.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
C. Berlato, M. A. Cassatella, I. Kinjyo, L. Gatto, A. Yoshimura, and F. Bazzoni
Involvement of Suppressor of Cytokine Signaling-3 as a Mediator of the Inhibitory Effects of IL-10 on Lipopolysaccharide-Induced Macrophage Activation
J. Immunol.,
June 15, 2002;
168(12):
6404 - 6411.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A.-K. Yi, J.-G. Yoon, S.-J. Yeo, S.-C. Hong, B. K. English, and A. M. Krieg
Role of Mitogen-Activated Protein Kinases in CpG DNA-Mediated IL-10 and IL-12 Production: Central Role of Extracellular Signal-Regulated Kinase in the Negative Feedback Loop of the CpG DNA-Mediated Th1 Response
J. Immunol.,
May 1, 2002;
168(9):
4711 - 4720.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
R. Lang, R. L. Rutschman, D. R. Greaves, and P. J. Murray
Autocrine Deactivation of Macrophages in Transgenic Mice Constitutively Overexpressing IL-10 Under Control of the Human CD68 Promoter
J. Immunol.,
April 1, 2002;
168(7):
3402 - 3411.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
H.-C. Wang and J. R. Klein
Multiple Levels of Activation of Murine CD8+ Intraepithelial Lymphocytes Defined by OX40 (CD134) Expression: Effects on Cell-Mediated Cytotoxicity, IFN-{gamma}, and IL-10 Regulation
J. Immunol.,
December 15, 2001;
167(12):
6717 - 6723.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
Y. Ding, L. Qin, D. Zamarin, S. V. Kotenko, S. Pestka, K. W. Moore, and J. S. Bromberg
Differential IL-10R1 Expression Plays a Critical Role in IL-10-Mediated Immune Regulation
J. Immunol.,
December 15, 2001;
167(12):
6884 - 6892.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. Sakatsume, Y. Xie, M. Ueno, H. Obayashi, S. Goto, I. Narita, N. Homma, K. Tasaki, Y. Suzuki, and F. Gejyo
Human Glomerulonephritis Accompanied by Active Cellular Infiltrates Shows Effector T Cells in Urine
J. Am. Soc. Nephrol.,
December 1, 2001;
12(12):
2636 - 2644.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
D. Deon, S. Ahmed, K. Tai, N. Scaletta, C. Herrero, I.-H. Lee, A. Krause, and L. B. Ivashkiv
Cross-Talk Between IL-1 and IL-6 Signaling Pathways in Rheumatoid Arthritis Synovial Fibroblasts
J. Immunol.,
November 1, 2001;
167(9):
5395 - 5403.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
R. Avdiushko, D. Hongo, H. Lake-Bullock, A. Kaplan, and D. Cohen
IL-10 receptor dysfunction in macrophages during chronic inflammation
J. Leukoc. Biol.,
October 1, 2001;
70(4):
624 - 632.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
R Hooghe, Z Dogusan, N Martens, B Velkeniers, and E L Hooghe-Peters
Effects of prolactin on signal transduction and gene expression: possible relevance for systemic lupus erythematosus
Lupus,
October 1, 2001;
10(10):
719 - 727.
[Abstract]
[PDF]
|
|
|
|
|
|
|
M. G. Schwacha, C. P. Schneider, K. I. Bland, and I. H. Chaudry
Resistance of macrophages to the suppressive effect of interleukin-10 following thermal injury
Am J Physiol Cell Physiol,
October 1, 2001;
281(4):
C1180 - C1187.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
C. J. Greenhalgh and D. J. Hilton
Negative regulation of cytokine signaling
J. Leukoc. Biol.,
September 1, 2001;
70(3):
348 - 356.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
N. Sekine, S. Fukumoto, T. Ishikawa, T. Okazaki, and T. Fujita
GH Inhibits Interferon-{gamma}-Induced Signal Transducer and Activator of Transcription-1 Activation and Expression of the Inducible Isoform of Nitric Oxide Synthase in INS-1 Cells
Endocrinology,
September 1, 2001;
142(9):
3909 - 3916.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. H. Dalpke, S. Opper, S. Zimmermann, and K. Heeg
Suppressors of Cytokine Signaling (SOCS)-1 and SOCS-3 Are Induced by CpG-DNA and Modulate Cytokine Responses in APCs
J. Immunol.,
June 15, 2001;
166(12):
7082 - 7089.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. Haque, H. Echchannaoui, R. Seguin, J. Schwartzman, L. H. Kasper, and S. Haque
Cerebral Malaria in Mice : Interleukin-2 Treatment Induces Accumulation of {{gamma}}{{delta}} T Cells in the Brain and Alters Resistant Mice to Susceptible-Like Phenotype
Am. J. Pathol.,
January 1, 2001;
158(1):
163 - 172.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
Q. Wang, Y. Miyakawa, N. Fox, and K. Kaushansky
Interferon-alpha directly represses megakaryopoiesis by inhibiting thrombopoietin-induced signaling through induction of SOCS-1
Blood,
September 15, 2000;
96(6):
2093 - 2099.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
T. Jinquan, C. Jing, H. H. Jacobi, C. M. Reimert, A. Millner, S. Quan, J. B. Hansen, S. Dissing, H.-J. Malling, P. S. Skov, et al.
CXCR3 Expression and Activation of Eosinophils: Role of IFN-{gamma}-Inducible Protein-10 and Monokine Induced by IFN-{gamma}
J. Immunol.,
August 1, 2000;
165(3):
1548 - 1556.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
D. Franchimont, J. Galon, M. Gadina, R. Visconti, Y.-J. Zhou, M. Aringer, D. M. Frucht, G. P. Chrousos, and J. J. O'Shea
Inhibition of Th1 Immune Response by Glucocorticoids: Dexamethasone Selectively Inhibits IL-12-Induced Stat4 Phosphorylation in T Lymphocytes
J. Immunol.,
February 15, 2000;
164(4):
1768 - 1774.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
Y. R. Boisclair, J. Wang, J. Shi, K. R. Hurst, and G. T. Ooi
Role of the Suppressor of Cytokine Signaling-3 in Mediating the Inhibitory Effects of Interleukin-1beta on the Growth Hormone-dependent Transcription of the Acid-labile Subunit Gene in Liver Cells
J. Biol. Chem.,
February 11, 2000;
275(6):
3841 - 3847.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. Grundstrom, M. Dohlsten, and A. Sundstedt
IL-2 Unresponsiveness in Anergic CD4+ T Cells Is Due to Defective Signaling Through the Common {gamma}-Chain of the IL-2 Receptor
J. Immunol.,
February 1, 2000;
164(3):
1175 - 1184.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
D. Krebs and D. Hilton
SOCS: physiological suppressors of cytokine signaling
J. Cell Sci.,
January 8, 2000;
113(16):
2813 - 2819.
[Abstract]
[PDF]
|
|
|
|
|
|
|
A. C. Ward, I. Touw, and A. Yoshimura
The Jak-Stat pathway in normal and perturbed hematopoiesis
Blood,
January 1, 2000;
95(1):
19 - 29.
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. P. Colgan, R. M. Hershberg, G. T. Furuta, and R. S. Blumberg
Ligation of intestinal epithelial CD1d induces bioactive IL-10: Critical role of the cytoplasmic tail in autocrine signaling
PNAS,
November 23, 1999;
96(24):
13938 - 13943.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. A. Cassatella, S. Gasperini, C. Bovolenta, F. Calzetti, M. Vollebregt, P. Scapini, M. Marchi, R. Suzuki, A. Suzuki, and A. Yoshimura
Interleukin-10 (IL-10) Selectively Enhances CIS3/SOCS3 mRNA Expression in Human Neutrophils: Evidence for an IL-10-Induced Pathway That Is Independent of STAT Protein Activation
Blood,
October 15, 1999;
94(8):
2880 - 2889.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
C.G. Begley and N.A. Nicola
Resolving Conflicting Signals: Cross Inhibition of Cytokine Signaling Pathways
Blood,
March 1, 1999;
93(5):
1443 - 1447.
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. Cottet, P. Dupraz, F. Hamburger, W. Dolci, M. Jaquet, and B. Thorens
SOCS-1 Protein Prevents Janus Kinase/STAT-dependent Inhibition of beta Cell Insulin Gene Transcription and Secretion in Response to Interferon-gamma
J. Biol. Chem.,
July 6, 2001;
276(28):
25862 - 25870.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
Top
Abstract
Introduction