Nri 2711

REVIEWS
The regulation of IL10 production by

immune cells
Margarida Saraiva* and Anne OGarra
Abstract | Interleukin10 (IL10), a cytokine with antiinflammatory properties, has a central

role in infection by limiting the immune response to pathogens and thereby preventing
damage to the host. Recently, an increasing interest in how IL10 expression is regulated in
different immune cells has revealed some of the molecular mechanisms involved at the
levels of signal transduction, epigenetics, transcription factor binding and gene activation.
Understanding the specific molecular events that regulate the production of IL10 will
help to answer the remaining questions that are important for the design of new strategies
of immune intervention.
*Microbiology and Infection

Research Domain, Life and
Health Sciences Research
Institute (ICVS), School of
Health Sciences, University of
Minho, Campus de Gualtar,
4710057 Braga, Portugal.
Division of Immunoregulation,
Medical Research Council
National Institute for Medical
Research, The Ridgeway,
Mill Hill, London NW7 1AA,
UK.
Correspondence to A.OG.
email:
[email protected]
doi:10.1038/nri2711
Published online
15 February 2010
The immune response has evolved to protect the host

from a wide range of potentially pathogenic microorganisms, but parallel mechanisms to control overexuberant immune responses and prevent reactivity to
self are required to limit host damage. Interleukin-10
(IL-10) is an anti-inflammatory cytokine with a crucial role in preventing inflammatory and autoimmune
pathologies13. IL-10-deficient mice4 develop inflammatory bowel disease following colonization of the
gut with particular microorganisms5 (BOX 1) and show
other exaggerated inflammatory responses to microbial challenge. Although the absence of IL-10 leads to
better clearance of some pathogens with no enhanced
immuno pathology 6,7, during other infections the
absence of IL-10 can be accompanied by an immunopathology that is detrimental to the host but does not
necessarily affect the pathogen load3,811. This suggests
that an absence of IL-10 is not always compensated by
other regulatory mechanisms and thus that there is a
non-redundant role for IL-10 in limiting inflammatory
responses in vivo.
To inhibit inflammatory pathologies, IL-10 functions at different stages of an immune response and
possibly at different anatomical locations. IL-10 was
initially described as a T helper 2 (TH2)-type cytokine12,
but further evidence suggested that the production of
IL-10 was associated with tolerant or regulatory T (TReg)
cell responses3,13,14. It is now known that the expression of IL-10 is not specific to TH2 cells or TReg cells but
instead that it is a much more broadly expressed cytokine
(FIG. 1). IL-10 is expressed by many cells of the adaptive immune system, including TH1, TH2 and TH17 cell
subsets, TReg cells, CD8+ T cells and B cells (reviewed in
ReFs 3,10,11,1416).
It is also expressed by cells of the

innate immune system, including dendritic cells (DCs),
macrophages, mast cells, natural killer (NK) cells, eosinophils and neutrophils3 (FIG. 1). Thus, IL-10 production
seems to be associated with many immune cells, affirming
its crucial role as a feedback regulator of diverse immune
responses, not only TH1 cell responses10,11 but also TH2
cell responses to schistosome parasites17, Aspergillus spp.18
and allergens19 (reviewed in ReF. 1).
Much is known about the function of IL-10. For
example, the induction of the anti-inflammatory
response mediated through the IL-10 receptor (IL-10R)
and activation of signal transducer and activator of
transcription 3 (STAT3) is reviewed in ReFs 3,20. By
acting on DCs and macrophages, IL-10 inhibits the
development of TH1-type responses (reviewed in ReF. 3)
but also leads to the suppression of TH2 cell and allergic responses (reviewed in ReF. 1). In addition to an
autocrine inhibitory effect of IL-10 on macrophages
and DCs, and because IL-10 can be produced by TH1,
TH2 and TH17 cells, an additional feedback loop exists
to limit the innate effector functions of macrophages
and DCs and their subsequent activation of T cells.
However, IL-10 enhances the differentiation of IL-10secreting TReg cells, thus providing a positive regulatory loop for its induction21 (reviewed in ReFs 1,14).
In some situations, IL-10 also activates mast cells and
enhances the functions of CD8+ T cells, NK cells and
B cells (reviewed in ReFs 2,3), although these effects
have yet to be tested in infection models.
So, IL-10 is a cytokine with important effects on the
development of an immune response. An understanding
of how IL10 expression is regulated in different innate
170 | MARCH 2010 | VoLuMe 10
www.nature.com/reviews/immunol
2010 Macmillan Publishers Limited. All rights reserved
REVIEWS
Box 1 | IL10 expression and gut homeostasis
The intestine is continuously exposed to bacterial flora, dietary antigens and
potential pathogens. To prevent chronic intestinal inflammation, various regulatory
lymphocyte populations keep the immune response in check. These populations
use several regulatory mechanisms, the best characterized of which involves
interleukin-10 (IL-10) and transforming growth factor- (TGF)15,155. IL-10- or IL-10
receptor-deficient mice do not develop severe autoimmune disorders but develop
colitis in the presence of microorganisms4,156. Many studies unequivocally identify
CD4+ T cell-derived IL-10 as a key mediator of intestinal immune homeostasis157161.
Coeliac disease and inflammatory bowel disease (IBD) are the most common causes
of non-infectious intestinal inflammation in humans, with recent reports identifying
IL10 as a susceptibility locus for the development of IBD162. Polymorphisms in
nucleotide-binding oligomerization domain 2 (NOD2) have also been associated
with IBD in humans163, which is interesting considering that NOD2 has been
associated with IL-10 production41. IL-10 seems to function not directly on T cells,
but instead on myeloid cell populations in a similar manner to that observed in the
immune response to pathogens3. In vivo IL-10 production by forkhead box P3
(FOXP3+) regulatory T (TReg) cells and FOXP3 regulatory T cells in the gut seems to
be mediated by TGF, independently of endogenous IL-10 (ReF. 97). This IL-10
independence is in contrast to that reported in vitro for human IL-10-producing
regulatory T cells14. Retinoic acid was identified as a cofactor for TGF in the
induction of FOXP3+ TReg cells164166, although retinoic acid itself downregulates the
expression of IL-10 by inducible FOXP3 regulatory T cells92. Although the exact
mechanisms of IL-10 induction in the intestine remain elusive, the protective role
of intestinal TReg cells mostly depends on their expression of IL-10, suggesting that
local IL-10 expression might be a therapy for IBD2.
Chromatin
Composed of nucleosomes,
this is the basic repeating unit
of eukaryotic genomes.
Nucleosomes consist of 146
base pairs of DNA wound
around an octamer of histone
proteins.
Plasmacytoid DC
A DC that lacks myeloid
markers such as CD11c and
CD33 but expresses high levels
of HLA-DR and CD123. These
cells produce high levels of
type I interferons in response
to viral infection.
and adaptive immune cells is therefore of importance

for the development of immune intervention strategies in various pathologies. Several layers of regulation of IL-10 expression exist, and this is a main focus
of this Review. First, regulation of IL-10 production
involves changes in the chromatin structure at the
IL10 locus. A second layer of regulation involves
the enhancement or silencing of IL10 transcription and
is controlled by specific transcription factors activated by
discrete signal-transduction pathways. In addition, posttranscriptional mechanisms exist. Many of the molecular events leading to IL10 expression are common to
various IL-10-producing immune cells. However, there
are also cell-specific signals and molecular mechanisms
that allow IL-10 production by particular immune cells
and not by others.
In this Review, we discuss our current understanding of the regulation of IL10 expression at the molecular
level in different cell types, from signal transduction
pathways to epigenetic regulation and the activation of
specific transcription factors involved in IL-10 production. Throughout, we highlight the common and distinct
mechanisms of IL-10 regulation that exist in different
IL-10-producing immune cells.
IL10 production by immune cells

Induction by pathogen-derived products. Pathogen
activation of DCs and macrophages involves the recognition of pathogen-derived products by pattern
recognition receptors (PRRs), which triggers the expression of cytokines and other factors 22. Both macrophages2327 and DCs26,2833 can express IL-10 in vitro
following activation of specific PRRs (FIG. 2a). In addition, DCs31,34, macrophages35 and neutrophils36 have been
reported to express IL-10 in vivo.
It has been suggested that Toll-like receptor 2 (TLR2)

agonists are specialized in inducing IL-10 expression
by antigen-presenting cells (APCs)29,30,37,38. For example, TLR2 signalling is crucial for the induction of
IL-10 production by macrophages (M. Teixeira-Coelho,
J. Carmona, A. G. Castro and M.S., unpublished observations) or by DCs39 stimulated with Mycobacterium
tuberculosis or with lipopeptides and the LcrV antigen
of Yersinia pestis 40. IL-10 production by macrophages
following pneumoccocal cell wall stimulation mainly
depends on TLR2; however, in this case a role for
nucleotide-binding oligomerization domain 2 (NoD2)
signalling, independent of TLR2, has also been
described 41. Significant amounts of IL-10 are also
produced by macrophages and myeloid DCs following
stimulation with TLR4 and TLR9 ligands 26. of note,
IL-10 production following TLR3 stimulation was only
observed in macrophages26. Interestingly, activation
of macrophages through TLRs results in high levels of
IL-10 production, whereas myeloid DCs only produce
intermediate amounts and plasmacytoid DCs (pDCs) do
not produce detectable levels of IL-10 (ReF. 26) (FIG. 1).
In addition, IL-10 can be induced by TLR-independent
stimuli, such as the C-type lectins DC-specific ICAM3grabbing non-integrin (DC-SIGN; also known as
CLeC4M)33 and dectin 1 (also known as CLeC7A)32
(FIG. 2a). Ligation of CD40 enhances IL-10 production by
TLR-stimulated28 or dectin 1-stimulated DCs32 and ligation of Fc receptors (FcRs) enhances IL-10 production
by TLR-stimulated macrophages25.
Signalling pathways for innate IL-10 production.
Following TLR ligation, signalling cascades are activated
through Toll/IL-1 receptor (TIR)-domain-containing
adaptor molecules, such as myeloid differentiation primary-response protein 88 (MYD88) and TIR-domaincontaining adaptor protein inducing IFN (TRIF; also
known as TICAM1), leading to the production of IL-10
and pro-inflammatory cytokines26,30,42. TLR signalling
through MYD88 leads to the activation of mitogenactivated protein kinases (MAPKs) and nuclear factor-B
(NF-B)43 (FIG. 2a).
Additional signals that are required for IL-10 production by macrophages have also been reported. of
note, optimal lipopolysaccharide (LPS)-induced IL-10
production by macrophages requires both the activation of the TRIF- and MYD88-dependent pathways26,27
and the production of and signalling by type I interferons (IFNs)27. This secondary induction of IL-10 by
type I IFNs has important implications for the use
of type I IFNs as potential anti-inflammatory drugs.
Moreover, this study is in line with the observation
that TNFR-associated factor 3 (TRAF3), an important
component of the type I IFN production pathway, is
also involved in LPS-induced upregulation of IL-10
expression42.
The MAPK cascade is composed of three major
groups of kinases: extracellular signal-regulated kinases
(eRKs) (comprising eRK1 (also known as MAPK3)
and eRK2 (also known as MAPK1), which are collectively referred to here as eRK); JuN N-terminal kinases
NATuRe ReVIewS | Immunology
VoLuMe 10 | MARCH 2010 | 171

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a
Low ERK expression
No IL-10 production
Plasmacytoid DC
Intermediate IL-10
production
Intermediate ERK
expression
Microbial
products
Myeloid DC
High ERK expression
High IL-10 production
Macrophage
High
antigen
dose
or IL-12
High antigen
dose and IL-12
IL-4
STAT4
ERK
STAT6
ERK
FOXP3+ TReg cell
DC
Naive
T cell
FOXP3 naive T cell

TGF and
retinoic acid
TGF, IL-6, IL-21

and/or IL-27
TGF
TGF
Signalling pathways?
STAT3
ERK
High antigen dose
TH1 cell
IL-12
IL-10-producing
TH1 cell
TH2 cell
IL-10
IL-10
IFN
TH17 cell
IL-10
IL-10-producing
FOXP3+ TReg cell
IL-10-producing
FOXP3 regulatory T cell
IL-10
IL-10
DC
Figure 1 | Interleukin10 expression in the immune system. a | Interleukin10 (IL10) is expressed by macrophages
and myeloid dendritic cells (DCs), but not by plasmacytoid DCs, in response to microbial products. The extracellular
Nature
| Immunology
signalregulated kinase 1 (ERK1) and ERK2 (which are collectively referred to here as ERK) pathway
is Reviews
one of the
signalling
cascades that is activated in these cells that results in IL10 expression. For other immune cells, such as B cells, mast cells
and eosinophils, the exact signalling pathways that lead to IL10 production remain elusive. b | In T helper (TH) cells, the
expression of IL10 is accompanied by the expression of the signature cytokines for each subset, with the exception of
regulatory T (TReg) cells, which normally lose the capacity to express other cytokines. Although the differentiation of TH
cells from naive CD4+ T cells requires T cell receptor triggering and the activation of distinct signal transducer and
activator of transcription (STAT) pathways, activation of the ERK pathway is a common requirement for IL10 expression by
these cells. High doses of antigen presented by DCs to naive T cells or IL12 favours the development of TH1 cells, which
produce interferon (IFN). IL10producing TH1 cells require high antigen dose and IL12 and STAT4 signalling for the
expression of maximum levels of IL10 following restimulation. In TH2 cells, IL4 and STAT6 signalling pathways are
required for IL10 expression. Induction of IL10producing TH17 cells is not well understood, but transforming growth
factor (TGF), IL6, IL21 and/or IL27 and STAT3 signalling are likely to be involved. c | TGF can induce the production
of IL10 by forkhead box P3 (FOXP3)+ TReg cells and this cytokine can also promote the development of IL10producing
FOXP3 regulatory T cells from naive T cells. Conversely, FOXP3+ IL10producing TReg cells can differentiate from naive
T cells in vitro in the presence of TGF and retinoic acid.
(JNKs) (comprising JNK1 (also known as MAPK8)

and JNK2 (also known as MAPK9)); and p38 (ReF. 44).
Following TLR stimulation, activation of eRK modulates
IL-10 expression30,4547, and in the presence of chemical inhibitors of eRK30,45,47 or in eRK-deficient cells46
IL-10 production by TLR-activated DCs is decreased.
Furthermore, the differences in IL-10 production by
macrophages, myeloid DCs and pDCs have been shown
to correlate with the strength of eRK activation in each
of these cell types47. Following TLR stimulation, eRK is
most highly activated in macrophages, with lower activation of eRK in myeloid DCs and the lowest amount of
activated eRK in pDCs47 (FIG. 1).
Further studies using cells deficient for tumour

progression locus 2 (TPL2) or NF-B1 (also known as
p105) support the role of eRK in the induction of IL-10.
TPL2 is an upstream activator of eRK and, following
TLR stimulation, TPL2 dissociates from the TPL2
NF-B1 complex and activates eRK. In the absence
of NF-B1, TPL2 is rapidly degraded in the cell and,
as a consequence, eRK activation by TPL2 is compromised48. In TPL2-deficient macrophages and myeloid
DCs the amounts of TLR-induced IL-10 were lower
than in wild-type cells owing to the absence of eRK
activation47. Similarly, NF-B1-deficient macrophages
have lower levels of IL-10 expression than control cells
172 | MARCH 2010 | VoLuMe 10
REVIEWS
a
Myeloid DCs
DC-SIGN
RAF1
Macrophages and myeloid DCs

Dectin 1
TLR2
NF-B
TLR2
TLR4
SYK
MYD88
ERK
TPL2
p38
IL-10
NOD2
RIP2
NF-B
TLR9
ERK
Cytoplasm
TLR2
MYD88
MEK1
MEK2
MSK1
MSK2
Macrophages
TLR3
Endosome
TRIF
IL-10
TRAF3
IL-10
TLR-dependent signal
Macrophage
IFN
TPL2
p38
IFN
ERK
PI3K
AKT
MSK1
MSK2
DUSP1
Through IL-10R
and STAT3
GSK3
CREB
AP1
IL-10
Through IL-10R
and STAT3
Figure 2 | Signals that induce interleukin10 expression by cells of the innate immune
response. a | The expression of interleukin10 (IL10) can be induced by Tolllike receptor
(TLR) or nonTLR signalling in macrophages and myeloid dendritic cells (DCs). Activation of
Nature Reviews | Immunology
TLRs and their adaptor molecules myeloid differentiation primaryresponse
protein 88
(MYD88) and TIRdomaincontaining adaptor protein inducing IFN (TRIF) results in
the activation of the extracellular signalregulated kinase 1 (ERK1) and ERK2 (which are
collectively referred to here as ERK), p38 and nuclear factorB (NFB) pathways.
Activation of these pathways results in the induction of IL10 expression, in addition to
proinflammatory cytokines. In myeloid DCs, nonTLR signals through DCspecific
ICAM3grabbing nonintegrin (DCSIGN) and RAF1 can augment TLR2induced IL10
production. Furthermore, activation of dectin 1 and the signalling molecules spleen
tyrosine kinase (SYK) and ERK results in IL10 production. In macrophages, a role for
nucleotidebinding oligomerization domain 2 (NOD2) signalling in IL10 induction, in
crosstalk with TLR2, has been described. b | Positive and negative feedback loops for IL10
regulation in macrophages. The p38 and ERK pathways leading to IL10 expression by
macrophages are tightly controlled by interferon (IFN) and IL10 itself. IL10 feeds back
to induce the expression of dualspecificity protein phosphatase 1 (DUSP1), which
negatively regulates p38 phosphorylation and thus limits IL10 production. IL10 can also
positively feed back to upregulate tumour progression locus 2 (TPL2) expression, thus
providing a positive amplification loop for its own production. In addition, IFN can also
interfere with the phosphoinositide 3kinase (PI3K)AKT pathway, releasing glycogen
synthase kinase 3 (GSK3). As GSK3 normally blocks IL10 expression by acting on the
transcription factors cAMP response elementbinding protein (CREB) and activator
protein 1 (AP1), IL10 production is inhibited by IFN through its effects on PI3K. IL10R,
IL10 receptor; MEK, MAPK/ERK1 kinase; MSK, mitogen and stressactivated protein
kinase; RIP2, receptorinteracting protein 2; STAT3, signal transducer and activator of
transcription 3; TRAF3, TNFRassociated factor 3.
following TLR activation49. IL-10 expression was only

partially restored following rescue of eRK activation in
these cells49, indicating that NF-B-mediated regulation
of IL-10 production involves both eRK-dependent and
eRK-independent mechanisms, as had been suggested
by previous studies50,51. Furthermore, pathogen triggering of DC-SIGN in human DCs resulted in the activation
of RAF1, leading to acetylation of the NF-B p65 subunit
and to prolonged and increased IL10 transcription52. This
effect was only observed after TLR-dependent NF-B
activation, suggesting that activation of DC-SIGN can
modulate TLR-induced IL-10 production52.
The regulation of IL-10 production in response to
dectin 1 ligation depends on spleen tyrosine kinase
(SYK)32. SYK is recruited to phosphorylated dectin 1
(ReF. 53) and initiates a signalling cascade that induces
IL-2 and IL-10 production32. IL-10 production downstream of dectin 1 also requires signalling through the
eRK pathway, despite being independent of TLR activation54 (FIG. 2a). Furthermore, IL-10 production by FcR
ligation in the presence of TLR signals in macrophages
can also lead to eRK activation55. Therefore, eRK activation is common to several signalling pathways upstream
of IL10 in macrophages and DCs.
IL-10 expression can also be compromised by inhibition of p38 signalling in LPS- or CpG-activated macrophages45,5658, primary DCs59 and human peripheral
blood monocytes60. Primary cells lacking the p38 regulator dual-specificity protein phosphatase 1 (DuSP1) have
prolonged p38 activation and increased levels of IL-10
expression following TLR stimulation6163. This could be
reversed by chemically inhibiting p38 signalling 61,62.
Interestingly, abrogation of either eRK or p38 activation leads to a reduction, but not abrogation, of IL-10
expression, which suggests that these two pathways might
cooperate in TLR-induced IL-10 production. Supporting
this hypothesis, inhibition of both the eRK and p38
pathways in LPS- or CpG-stimulated macrophages leads
to an almost complete abrogation of IL-10 production
(A.oG., unpublished observations). Furthermore, deficiency of mitogen- and stress-activated protein kinase 1
(MSK1; also known as RPS6K5) and MSK2 (also known
as RPS6K4), which are activated downstream of the p38
and eRK pathways, correlated with a loss of IL-10 expression by LPS-stimulated macrophages64.
The production of IL-10 by macrophages and DCs
is also regulated by the activation of certain inhibitory
pathways. eRK- and p38-dependent IL-10 production
is inhibited by IFN38 (FIG. 2b). In addition to directly
blocking TLR-induced MAPK activation, IFN induces
the release of glycogen synthase kinase 3 (GSK3) by
antagonizing phosphoinositide 3-kinase (PI3K)AKT
activation. This leads to inhibition of TLR-induced
IL-10 production by suppressing the binding of activator protein 1 (AP1) to the Il10 promoter 38. Another
negative feedback loop controlling IL-10 production by
macrophages is mediated by IL-10 itself. IL-10 induces
the expression of DuSP1, which negatively regulates p38
phosphorylation and thus limits IL-10 production65. By
contrast, IL-10 positively feeds back to upregulate Tpl2
expression66, thus providing a positive amplification
VoLuMe 10 | MARCH 2010 | 173

REVIEWS
loop for its own production. IL-10 was also described
to induce its own transcription in human monocytes in
a STAT3-dependent manner 67, which may result from
its upregulation of TPL2 and thus eRK activation66.
However, the mechanisms dictating the balance between
IL-10-mediated negative and positive feedback loops are
currently not clear. Furthermore, the inhibition of IL-10
production by NK cell- or T cell-derived IFN in vivo
will also influence these loops.
Various pathogen-derived products induce IL10
expression by macrophages and DCs through the activation of signalling cascades that, although common
to various stimuli and different cells, have distinct
thresholds of activation depending on the cell type,
which reflect the distinct amounts of IL-10 produced
by these cells.
IL-10 production by TH cells. IL-10 production was first
described in TH2 cells12,68, where its expression accompanies that of the TH2-type cytokines IL-4, IL-5 and
IL-13. TH1 cells can also be induced to produce IL-10,
but, in contrast to TH2 cells, only under certain conditions10,11,15,6976 (FIG. 1). Furthermore, TH17 cells have
recently been shown to produce IL-10 (ReFs 72, 7779).
The fact that TH1, TH2 and TH17 cells are dependent on
DC- and macrophage-derived factors that are downregulated by IL-10, but these subsets can all be induced to
produce IL-10, is indicative of a negative feedback loop
that ensures that effector T cell responses do not result
in immunopathology. It is of interest to note that IL-9producing TH cells, which have recently been suggested to
be a unique TH cell subset (TH9 cells), also express IL-10
(ReF. 80).
Notch
A signalling system
comprising highly conserved
transmembrane receptors that
regulate cell fate choice in the
development of many cell
lineages. Therefore, they are
crucial in the regulation of
embryonic differentiation
and development.
Molecular signals for IL-10 induction in T H cells.

IL-10-inducing signalling cascades have been studied
less thoroughly in TH cells than in macrophages and
DCs. IL-10-producing TH1 cells have been described
in infectious diseases, human CD4+ T cell clones and
mouse CD4+ T cells. TH1 cells that produce both IFN
and IL-10 can be generated by inducing T cells to proliferate with high levels of antigen-specific or polyclonal
stimulation in the presence of IL-12 (ReFs 10,11,69,70)
(FIG. 1). However, until recently the signals that determine
whether TH1 cells produce IL-10 were not known. Strong
T cell receptor (TCR) triggering (high antigen dose)71 and
endogenous IL-12 have now been shown to be essential
for the differentiation of IL-10-producing TH1 cells, as
well as for maximal expression of IL-10 following restimulation of these cells72. IL-10 induction in TH1 cells
is STAT4 and eRK dependent 72 (FIG. 1). Notch signalling
can also induce IL-10 expression by TH1 cells, a process
that requires STAT4 (ReF. 81). In TH2 cells, IL-10 production seems to be regulated by the main TH2 type-associated signalling pathways and transcription factors: IL-4,
STAT6 and GATA binding protein 3 (GATA3)8284. IL-10
expression by TH17 cells seems to occur in a STAT3- and,
in some cases, STAT1-dependent manner 79,85 (FIG. 1).
Thus, to induce IL-10 expression, TH1, TH2 and TH17
cells require the same signals needed for each TH cell
differentiation programme. However, IL-10 production
by all these subsets requires eRK activation72, indicating

that a common molecular mechanism exists for IL-10
production by TH cells. Chemical inhibition of the p38
signalling pathway did not compromise the production
of IL-10 by TH1, TH2 or TH17 cells72, suggesting that in TH
cells the role of eRK is dominant over that of p38. This
observation is in contrast to a joint role for eRK and p38
in IL-10 induction in macrophages and DCs.
IL-21 can enhance IL-10 expression by CD4+ T cells
in the context of different stimuli86, and IL-27 enhances
IL-10 expression by TH1, TH2 and TH17 cells78,79,85,87,88.
By contrast, IL-27 attenuates TLR-induced IL10 expression by human monocytes89. of interest, it has recently
been shown that both IL-21 and IL-27 induce eRK
activation90,91, but it is currently not clear whether this
explains the ability of these cytokines to upregulate IL-10
production. Also, both IL-21 and IL-27, in addition to
eRK, activate STAT3, which seems to be involved in
IL-27-mediated IL-10 upregulation by T cells79.
undoubtedly, all T cell subsets can produce IL-10, as
well as their hallmark cytokines, following TCR triggering, but this depends on the environmental context and
strength of stimulus10,11,6973,9294.
IL-10 and regulatory T cells. TReg cells, which are characterized by their specific expression of the transcription factor forkhead box P3 (FoXP3), do not express
IL-10 following stimulation directly after ex vivo isolation95,96, unless isolated from the gut 97 (BOX 1). Although
FoXP3+ TReg cells inhibit naive T cell proliferation in vitro
independently of IL-10, in some cases, TReg cells mediate
their regulatory function in vivo through IL-10 (reviewed
in ReFs 1,14,16,98100). Therefore, TReg cells must receive
signals in vivo to induce the expression of this suppressive
cytokine. Both IL-2 and IL-4 have been shown to induce
IL-10 production after culture of TReg cells in vitro101,102.
However, in these studies, the TReg cell population analysed
might have contained some effector T cells and therefore
the source of IL-10 cannot be confirmed. So far the signals
that induce IL-10 expression by FoXP3+ TReg cells remain
elusive, although transforming growth factor (TGF)
has been shown to be required in vivo97 (FIG. 1).
Several populations of antigen-driven FoXP3
IL-10-producing T cells with regulatory activity that are
distinct from naturally occurring TReg cells have been
described (reviewed in ReFs 1,2,13,14). These cells produce IL-10, but not IL-2, IL-4 or IFN, and can be generated in vitro using various stimuli, such as cytokine
cocktails (TGF, IL-10 and IFN) or immunosuppressive drugs (vitamin D3 and dexamethasone)1,2,13,14,21, or
in vivo by repeated stimulation with soluble antigen71,103.
Additional signals for IL-10 expression by these FoXP3
regulatory T cells include co-stimulation through CD2
or CD46 and stimulation with type I IFNs or with
immature DCs (reviewed in ReFs 1,14,104). Signals
delivered through inducible T cell co-stimulator (ICoS)
have also been suggested to induce IL-10 expression by
FoXP3 regulatory T cells105,106; a similar effect has been
observed in TH2 cells107,108, suggesting that, although it
is involved in the induction of IL-10, ICoS is not a cell
type-specific inducer of IL-10.
174 | MARCH 2010 | VoLuMe 10
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DNaseI hypersensitive sites
sites of nuclease sensitivity in
the nuclei on exposure of cells
to limiting concentrations of
DNaseI. The digested regions
of DNA correspond to sites of
open DNA, which might be
factor-binding sites or areas of
altered nucleosome
conformation.
Chromatin remodelling
Alterations that are induced
in chromatin by enzymes
that modify the extent of
acetylation, methylation or
other covalent modifications
of histones.
Acetylation
A post-translational
modification of chromatin
components, particularly
histones. It correlates with
actively transcribed chromatin.
It will be of interest to determine whether IL-10producing FoXP3 regulatory T cells differentiate directly
from naive T cells or are derived from TH1, TH2, TH9
or TH17 cells that have lost expression of their effector T cell cytokines but have maintained IL-10 expression. It is possible that FoXP3 regulatory T cells that
only make IL-10 have originally differentiated along a
TH1 cell pathway through repeated high-level antigenic
stimulation, which results in IL-10 production and ultimately in the downregulation of TH1 cell production
of IFN by feedback inhibition of IL-12 production by
DCs and macrophages71.
Additional cell types that produce IL-10. In addition
to macrophages, DCs and CD4+ T cells, other cells of
the immune system are also known to express IL-10.
CD8+ T cells express IL-10 following TCR activation
or interaction with CD40 ligand expressed by activated
pDCs109111, and this IL-10 production can be enhanced
by IL-21 (ReF. 86). Stimulation of B cells with autoantigens, TLR4 and TLR9 ligands or vitamin D3 also
leads to IL-10 production112117. Finally, mast cells can
express IL-10 following TLR4 activation or during skin
allergic or damage responses1,118,119. Recently, neutrophils were reported to produce IL-10 in response to TLR
and C-type lectin co-activation through MYD88 and
SYK, respectively 36. These IL-10-producing neutrophils
were shown to be recruited to the lung during mycobacterial infections and to regulate local immune inflammatory responses36. It is currently not clear whether
the molecular mechanisms required for the induction
of IL-10 by these cells are regulated by the common
factors that regulate IL-10 production by T H cells,
macrophages and DCs.
Box 2 | Epigenetic control of IL10 expression

Several studies suggest that the expression of interleukin-10 (IL-10) is regulated by
changes in the structure of the chromatin at the IL10 locus51,133135 (FIG. 3). Various
DNaseI hypersensitive sites (HSSs) were found in the mouse Il10 locus, most of which are
common to IL-10-producing T cells, macrophages and dendritic cells (DCs). However, a
macrophage-specific regulatory element (HSS4.5), which is absent in T cells, was also
found51. Although chromatin remodelling seems to be one of the initial events leading to
IL10 expression, additional signals are required to allow high rates of IL10 transcription.
Epigenetic imprinting of the Il10 locus in mice, as measured by histone acetylation, was
observed in high-IL-10-producing T helper 2 (TH2 cells)84 and macrophages51 but not in
low-IL-10-producing TH1 cells84, despite the open conformation of the IL10 locus
observed for all these cell types. In macrophages, the histones at the Il10 locus were also
reported to be hyperphosphorylated55,167. Furthermore, by interacting with the distal
segment of the IL10 promoter, histone deacetylase 11 negatively regulated the
expression of this cytokine in human and mouse antigen-presenting cells168.
Several studies have identified GATA binding protein 3 (GATA3) as a possible
initiator of chromatin remodelling and histone acetylation of the Il10 locus in
mice83,133. However, as GATA3 is only expressed by TH2 cells and not in other
IL-10-producing TH cells, macrophages or DCs, other mechanisms must operate in
these cells to induce chromatin remodelling at the Il10 locus. A clue may come
from the finding that, in macrophages, remodelling of the Il10 locus occurs following
TLR stimulation51 or Fc receptor binding in an extracellular signal-regulated kinase
(ERK)-dependent manner55.
Overall, analysis of the chromatin conformation at the IL10 locus may help to explain
the different levels of IL10 expression and the different factors involved in this
expression by different cell types, although many questions still remain unanswered.
understanding the molecular pathways leading

to IL-10 production by different immune cells might
provide valuable information on possible targets for
IL-10 manipulation. This will be useful in the design
of intervention strategies to modulate IL-10 production
and ultimately the immune response.
Transcription factors that regulate IL10

Activating the IL10 promoter. The structure of the human
and mouse IL10 promoters is similar and both contain a
TATA box and a CCAAT box (CCAGT in mice). The
human and mouse IL10 promoters have a high level of
homology, particularly around certain putative binding
sites for transcription factors. However, the presence of a
conserved putative binding site in a gene promoter does
not guarantee transcription factor binding.
In addition to epigenetic control (BOX 2), the expression
of IL10 depends on transcription factor binding. The transcription factors specific protein 1 (SP1)120, SP3 (ReF. 121),
CCAAT/enhancer binding protein- (C/eBP)122,123, IFNregulatory factor 1 (IRF1) and STAT3 (ReF. 124) have been
proposed to bind to and transactivate IL10 in macrophage
and T cell lines of mouse or human origin (FIGs 3, 4). Also,
binding of the NF-B p50 subunit to the IL10 promoter in
a human T cell lymphoma cell line has been described125.
Moreover, some of these findings depend on the cell type
and on the stimulus used. For example, whereas one study
indicates that IL10 promoter activity relies on an SP1 site
located between positions 636 and 631 relative to the
initiation site56, another report shows that IL10 promoter
activity relies on the C/eBP5 motif positioned between
the TATA box and the translation start point122. Although
both studies were carried out using the human promonocytic cell line THP1, the type of stimulation was
different (LPS versus cyclic AMP (cAMP), respectively),
which might account for the differences observed.
Studies using mouse primary cells have shown that
homodimers of NF-B p50 bind to the proximal Il10
promoter, activating Il10 transcription in primary macrophages126, and p50-deficient macrophages have an
impaired expression of IL-10 following LPS stimulation
compared with wild-type macrophages126. Furthermore,
IL-10 induction in response to double-stranded RNA
stimulation and viral infection of mouse macrophages
was described to be protein kinase R (PKR) dependent and to be regulated by binding of NF-B to a distinct site in the Il10 promoter 127. In addition to NF-B,
a role for C/eBP in cAMP-mediated IL-10 production was confirmed in mouse primary macrophages;
nuclear accumulation and DNA binding of C/eBP was
involved in IL-10 production in response to adenosine
and Escherichia coli infection, and C/eBP-deficient
macrophages failed to produce IL-10 (ReF. 128).
Recently, two cofactors of the homeobox (HoX) family, pre-B-cell leukaemia transcription factor 1 (PBX1)
and PBX-regulating protein 1 (PReP1), were implicated
in inducing IL-10 expression by mouse macrophages129.
In this study, the expression of IL-10 was triggered by
the interaction of macrophages with apoptotic cells
and depended on p38; in human cells, transcription
of IL10 cells was mediated by the binding of PBX1 and
VoLuMe 10 | MARCH 2010 | 175

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IL10 locus
Stronger
in TH2 cells
Weaker in
TH1 cells
Only in macrophages
and DCs
Only in
HSS4.5
undifferentiated
CD4+ T cells
HSS8.8
Enh
HSS9.1
TH2 cells
HSS0.12
HSS+0.92
HSS+2.98
HSS+6.45
HSS+1.65
1 kb
Enh
JUN
NF-B
IL10 promoter
Enh
Enh
SMAD4
TH1 cells
NF-B
Enh
GATA3 STAT MAF
TH2 locus
remodelling
Macrophages
and TH17 cells
Enh
Enh
Enh
Enh
C/EBP
CREB
ATF1
SP1
SP3
STAT NF-B TATA box
Macrophages
Macrophages
Enh
Start
site +1
Macrophages
Macrophages
and T cell lines
Macrophages
Figure 3 | molecular regulation of interleukin10 expression: the interleukin10 locus and promoter. The mouse
interleukin10 (Il10) locus (top panel) and Il10 promoter (bottom panel) are represented here. Several
DNaseI
hypersensitive
Nature
Reviews
| Immunology
sites (HSSs), and their relative position to the Il10 starting site (+1), are indicated. Most of these HSSs are common to all cells,
although some cellular specificity is also observed. Two of these sites (HSS4.5 and HSS+6.45) have been studied in more detail,
and their role in Il10 regulation has been described. HSS4.5 contains hyperacetylated histones and binds nuclear factorB
(NFB) in macrophages, whereas HSS+6.45 binds JUN proteins in T helper 2 (TH2) cells. Both HSS4.5 and HSS+6.45 were
shown to enhance the Il10 promoter activity in reporter assays. The biological role for the other HSSs needs to be further
clarified. The proximal elements that regulate the expression of IL10, including the Il10 promoter, have been well studied.
Several transcription factors have been shown to bind to the Il10 promoter and to enhance Il10 transcription in various cell
types. In TH2 cells, GATA binding protein 3 (GATA3) functions as a master regulator for Il10 expression by binding to sites in the
Il10 locus (including to the promoter) and inducing locus remodelling. Also represented are putative signal transducer and
activator of transcription (STAT) binding sites in the mouse Il10 promoter. ATF1, activating transcription factor 1; C/EBP,
CCAAT/enhancer binding protein; CREB, cAMPresponsiveelementbinding protein; DC, dendritic cell; Enh, enhancer;
GATA3, GATA binding protein 3; SMAD4, mothers against decapentaplegic homologue 4; SP, specific protein.
PReP1 to the apoptotic cell-response element (ACRe) in

the IL10 promoter 129. The transcription factors cAMPresponsive-element-binding protein (CReB) and activating transcription factor 1 (ATF1) have been shown
to be activated by MSKs in LPS-stimulated mouse
macrophages and to bind to the Il10 promoter, thus suggesting a direct effect of the kinases MSK1 and MSK2 in
the regulation of IL-10 induction64 (FIG. 2b). There is also
evidence for TGF1-induced SMAD4 binding to and
activating the Il10 promoter in mouse T H1 cells 130;
however, it is worth noting that TGF1 inhibits the
development of TH1 and TH2 cells in vitro and thus their
production of IL-10 (ReF. 72).
The transcription factor GATA3 was shown to be a
master regulator of IL-10 expression in mouse TH2 cells by
binding to and initiating changes in the chromatin structure at the Il10 locus83,84. Although one of the binding sites
for GATA3 is located in the Il10 promoter, GATA3 alone
does not transactivate the Il10 promoter 83. GATA3 may
thus be responsible for remodelling the Il10 locus in TH2
cells, with other factors being necessary to induce high levels of IL-10 expression in other cell types such as TH1 cells
that do not express GATA3 (ReFs 69,72).
originally described as a TH2 cell-specific factor 131,
the transcription factor MAF has been shown to bind
to the Il10 promoter and have a role in the transcriptional
regulation of IL-10 in mouse macrophages stimulated
with LPS and IL-4, although MAF alone is not sufficient
to induce Il10 expression in these cells132. other recent

reports have also implicated MAF in the expression of
IL-10 by TH17 cells, showing binding of this transcription factor to MAF recognition elements in the Il10
promoter 85, and in the differentiation of IL-10-producing
FoXP3 regulatory T cells88. Finally, MAF expression is
also detectable in TH1, TH2 and TH17 cells, correlating
with IL-10 production, and MAF expression depends
on eRK activation in TH1 and TH17 cells, similar to
IL-10 expression72. Taken together, these reports suggest that MAF may be a universal transcription factor
for the regulation of IL-10 production, important in all
IL-10-producing cells of both the innate and the adaptive
immune systems.
As discussed, various transcription factors have been
described to bind to and activate the IL10 promoter
(FIGs 3, 4). The activity of one transcription factor or
another to regulate IL10 expression seems to depend on
the cell type and the type of stimuli. Furthermore, multiple studies suggest that IL-10 is not regulated in a simple
and linear manner, which is in accord with its function
to keep diverse immune responses in check.
Enhancing IL10 transcription. Following the description
of several distal regulatory elements in IL10 51,133135 (FIG. 3),
the search for transcription factors with a role in regulating IL10 expression has been expanded to those that bind
regions of the locus outside of the promoter region.
176 | MARCH 2010 | VoLuMe 10
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Macrophages
p38
TPL2
NF-B
SP1
C/EBP
ATF1
ERK
MAF
CREB
STAT4
MAF*
SMAD4
PREP1
PBX1
IL-10
GATA3
ERK
TH2 cells
JUN
MAF* STAT6
MAF
TPL2
DCs
ERK
NF-B
NF-B
p38
TH1 cells
ERK
STAT3
SYK
RAF1
NF-B
ERK
TH17 cells
Figure 4 | Transcription factors that control interleukin10

expression by CD4+ T cells and antigenpresenting cells.
Reviews
| Immunology
Many transcription factors haveNature
been found
to regulate
the
expression of interleukin10 (IL10) both in antigen
presenting cells and in CD4+ T cells. Represented here are
the signalling molecules (in the outer circle) and transcription
factors (inner circle) involved in IL10 regulation with a role
validated by promoter studies (mutagenesis or chromatin
immunoprecipitation) or by studies in genetically modified
mice. Part of these studies were carried out in cell lines,
although recently the study of IL10 regulation has involved
primary cells. Some of the indicated transcription factors are
cell specific and others seem to have a wider role, as discussed
in detail in the text. The transcription factors in bold are
common to various cell types in regulating IL10 production.
Transcription factors marked with * are those that have not
been shown to bind the promoter or the Il10 locus. ATF1,
activating transcription factor 1, C/EBP, CCAAT/enhancer
binding protein; CREB, cAMPresponsiveelementbinding
protein; DC, dendritic cell; ERK, extracellular signalregulated
kinase; GATA3, GATA binding protein 3; NFB, nuclear
factorB; PBX1, preBcell leukaemia transcription factor 1;
PREP1, PBXregulating protein 1; SMAD4, mothers against
decapentaplegic homologue 4; SP1, specific protein 1; STAT,
signal transducer and activator of transcription; SYK, spleen
tyrosine kinase; TH, T helper; TPL2, tumour progression locus 2.
The NF-B p65 subunit binds to a newly described

B site located 4.5 kb upstream of the Il10 start site
and has a role in enhancing IL-10 expression by LPSstimulated mouse macrophages 51 (FIG. 3). This is in
keeping with a report that mice deficient for inhibitor of NF-B kinase 2 (IKK2) show a defect in IL-10
production in LPS-stimulated macrophages 50. This
B site is exposed in IL-10-producing LPS-, CpG- or
zymosan A-stimulated mouse macrophages and DCs
but is absent in T cells51. This suggests that different
molecular mechanisms might regulate the expression
of Il10 in the innate versus adaptive immune systems
and that the production of IL-10 by innate immune cells
might be subject to additional regulation, as it is the first
checkpoint for the initiation of immune responses and
determines the class of the resulting adaptive immune
response. of interest, however, is that eRK signalling is
required for optimal IL-10 induction in macrophages,
DCs and TH cell subsets, which questions whether
transcription factors that are required for IL-10 induction in DCs and macrophages may also have a role in
inducing IL-10 in T cells. one of the transcription factors implicated in TLR-induced IL-10 expression in
mouse macrophages and DCs, by the activation of eRK,
is FoS, the expression of which is strongly induced by
high levels of eRK activation29,30,47.
Studies have suggested there is a role for JuN proteins
in regulating Il10 in mouse TH2 cells, but not TH1 cells,
through binding to a regulatory element located ~6.45 kb
downstream of the Il10 start site133,134 (FIG. 3). This finding supports our suggestion that alternative mechanisms
probably operate to regulate Il10 induction in TH1 and
TH2 cells and possibly in other TH cell subsets, although
common factors, such as eRK and MAF, are required for
Il10 induction in various cell types.
Although there are no data so far to show direct
binding of the various STATs to the IL10 locus, an
increasing amount of evidence suggests a role for the
STAT proteins in regulating the induction of IL-10
expression in both primary macrophages and T cells.
In mouse T cells, the induction of IL-10 by IL-27
seems to depend on both STAT1 (ReFs 79,87) and
STAT3 (ReFs 79,85), and STAT3 is also involved in IL-6induced IL-10 expression79. By contrast, a recent study
on human monocytes describes an inhibitory role for
IL-27 on IL-10 production through STAT1 (ReF. 89). In
addition, as STAT3 is required for TH17 cell differentiation, this transcription factor may have an indirect
role in IL-27-mediated induction of IL-10 by T H17
cells by modulating TH17 cell differentiation. Another
study suggests that IL-10 induces its own expression
by human monocyte-derived macrophages in an
autocrine manner through the activation of STAT3
(ReF. 67). In this study, activation of the IL10 promoter
depended on the integrity of the STAT3-binding site.
Considering the data from human cell lines that show
binding of STAT3 to the IL10 promoter 124, it is possible that the STAT3-dependent effects on IL10 activation in primary cells might be related to promoter
transactivation. Finally, STAT4, which is important
for the differentiation of IFN- and IL-10-producing
mouse TH1 cells72, was also reported to have a role in
inducing IL-10 expression by mouse NK cells136. The
molecular mechanisms underlying the participation of
other STAT molecules in the control of IL-10 expression require further clarification, particularly as several
STAT molecules are required for the differentiation of
TH cell subsets and thus can modulate the induction
of IL-10 in an indirect manner.
The list of transcription factors involved in the regulation of IL10 expression is expanding (FIG. 4), which
reflects the degree of precision and complexity that the
expression of this cytokine demands. The exact contribution of many of the transcription factors discussed
above remains elusive and may in some cases be cell specific, but may also depend on the type of stimulus that
triggers IL10 expression or may affect IL10 expression
indirectly. with the identification and characterization
of distal regulatory regions in the IL10 locus, a role for
other transcription factors might also be revealed.
VoLuMe 10 | MARCH 2010 | 177

REVIEWS
T follicular helper cell

(TFH cell). A CD4+ T cell that
provides help to B cells in
follicles and germinal centres.
The TFH cell signature includes
the expression of CXCR5, ICOs,
CD40 ligand and IL-21, factors
that mediate TFH cell homing to
follicles and B cell help.
MicroRNAs
single-stranded RNA
molecules of approximately
2123 nucleotides in length
that regulate the expression
of other genes.
Silencing IL10 expression. Recent studies have provided

evidence for the role of certain transcription factors in
silencing Il10 expression. For example, it has been suggested that the transcription factor eTS1 has a role in
repressing the production of IL-10 by mouse TH1 cells,
as eTS1-deficient TH1 cells show a marked increase in
the production of this cytokine137. However, it is possible
that the effect of eTS1 on IL-10 expression results from
diminished TH1 cell differentiation, as no interaction of
eTS1 with the Il10 locus has been shown. Similarly, an
increase in IL-10 expression was observed in mice deficient for the TH1 cell-specific transcription factor T-bet
(also known as TBX21) that were infected with M. tuberculosis, suggesting that T-bet might have a role in the
negative regulation of IL-10 expression by TH1 cells138.
However, as IFN expression is also lost in the absence of
T-bet, this effect on IL-10 could reflect a blockade of TH1
cell differentiation, with the increase in IL-10 expression
resulting from other cells. This notion is strongly supported by the observation that IL-10 production by TH1
cells is accompanied by the expression of high levels of
IFN and T-bet 69,72.
Silencers of IL-10 expression have also been identified in cells of the innate immune response. MHC class II
transactivator (CIITA) has been shown to negatively regulate the expression of IL-10 by mouse DCs and the activity
of the Il10 promoter in a mouse macrophage cell line139.
As mentioned above, it is also possible that STAT1 negatively regulates IL-10 expression in human monocytes89,140.
However, it is still unclear whether STAT1 directly or indirectly affects the Il10 locus. In addition, isolated peritoneal
macrophages from B cell lymphoma 3 (BCL-3)-deficient
mice produce increased amounts of IL-10, suggesting that
BCL-3 is an inhibitor of IL-10 expression in macrophages;
although, again, it is questionable whether this effect is
direct or indirect 141. BCL-6 has been reported to have
a role in inhibiting the production of TH2 cell-specific
cytokines including IL-10 (ReF. 142), and in its absence
T cells activated with strong co-stimulation show a large
upregulation of Il4, Il10 and Il13 mRNA, whereas overexpression of BCL-6 in wild-type T cells strongly inhibits
the production of IL-10 following activation142. However,
it is unknown whether BCL-6 directly regulates the Il10
locus. In recent reports, BCL-6 was required for T follicular
helper cell differentiation, but it inhibited the differentiation of other TH cell subsets, by direct interaction with
T-bet, GATA3 and retinoic acid receptor-related orphan
receptor-t (RoRt)143145. It is therefore possible that the
previously reported BCL-6-mediated IL-10 suppression in
TH2 cells142 is a consequence of the inhibition of the TH2
cell differentiation pathways and therefore not due to a
direct effect of BCL-6 on Il10 transcription. In support of
this, no BCL-6 consensus binding sites have as yet been
found in the Il10 promoter 142.
The complexity of IL10 regulation by different
cells of the immune system, having both positive and
negative feedback loops, shows the tight control that
is essential to achieve a balance between an effective
immune response and immunopathology. This complex
regulation ranges from common to distinct pathways of
IL-10 induction in different cell types.
Mechanisms of posttranscriptional regulation

Modulation of mRNA stability is an important component in the regulation of expression of several cytokines
(reviewed in ReF. 146) and most cytokine genes have a
long 3 untranslated region (uTR), containing class II
adenosineuridine-rich elements (ARe) that target
mRNAs for rapid degradation. Multiple copies of potential mRNA destabilizing motifs are found in the 3 uTR
of Il10 mRNA147. Various factors can alter the stability of
Il10 mRNA, including IL-10 itself, which triggers Il10
mRNA degradation148,149, and adenosine receptor activation, which acts by relieving the translational repressive
effect of the Il10 3 uTR thereby increasing the mRNA
half-life and the amount of IL-10 produced150.
More recently, Il10 mRNA has been identified as a
tristetraprolin (TTP) target in a wide genome screen151.
TTP is a RNA-binding molecule that can induce rapid
degradation of mRNA following binding to ARes in
3 uTRs. Supporting this finding, macrophages from
TTP-deficient mice showed a decrease in the rate of
Il10 mRNA decay and an increase in IL-10 secretion151.
Moreover, activation of p38 has been reported to stabilize the Il10 mRNA by inhibiting the action of TTP152.
Interestingly, IL-10 induces TTP expression in a STAT3dependent manner, contributing to the establishment of
an anti-inflammatory programme153.
Finally, a role for microRNAs in the regulation of IL-10
expression has been described134. A recent report shows
that the human microRNA miR-106a, expressed in cells
of both lymphoid and myeloid origin, binds the 3 uTR of
the IL10 mRNA and induces its degradation154.
This level of post-transcriptional regulation of IL-10
expression might explain why, despite the existence of
common pathways for IL-10 induction, different cells ultimately secrete different amounts of IL-10. Thus, in addition to genetic regulation, post-transcriptional regulation
contributes to the fine tuning of IL-10 expression.
Conclusion and outstanding questions
owing to the key role of IL-10 in the immune response
and the link between defective IL-10 production and
certain autoimmune and inflammatory diseases, an
understanding of the molecular mechanisms that regulate the expression of this cytokine is crucial.
The fact that various cell types can express IL-10 makes
the subject of IL10 regulation challenging. It also highlights the complexity of this regulation. Several early studies on the molecular regulation of IL-10 reported apparent
differences. The fact that cell lines and different conditions
were used in most of these studies has certainly been a
contributing factor to these discrepancies. Indeed, recent
studies using primary T cells, macrophages and DCs show
more consistent results between laboratories with respect
to the regulation of IL10 expression.
Several general conclusions from these studies
can be made: first, many cells of the innate and adaptive immune response produce IL-10 regardless of the
stimulus. Second, different stimuli, or the strength of
the stimulus, give rise to different levels of IL-10 in the
same cell type. Third, some of the molecular mechanisms
for the regulation of IL-10 differ according to the cell
178 | MARCH 2010 | VoLuMe 10
REVIEWS
type, although common mechanisms also exist. And
fourth, IL-10 is induced in many situations together with
pro-inflammatory cytokines, although the pathways that
induce IL-10 expression may actually negatively regulate
the expression of these pro-inflammatory cytokines.
Several outstanding questions and future challenges
remain. which cells are induced to produce IL-10 during
an immune response to specific pathogens and gut flora,
and which IL-10-producing cells are required to prevent
host damage or to conversely inhibit immune responses,
thereby contributing to chronic infection? what signalling pathways and transcription factors can specifically
induce IL-10 in different immune cells independently
of the induction of pro-inflammatory cytokines? what
signalling pathways are required in different cells, and
what is the hierarchy of transcription factor binding to
IL10 regulatory elements? Is IL-10 production in vivo
dictated by the environment and inflammatory stimuli, and what maintains the remodelling of the IL10
locus? For example, can TH1 cells detect signals and
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Acknowledgements
We thank L. Gabrysova for critical reading of and commenting

on this review and A. Howes for careful proof reading.
Competing interests statement
The authors declare no competing financial interests.
DATABASES
UniProtKB: http://www.uniprot.org
CIITA | dectin 1 | DUSP1 | ETS1 | IL10 | MAF | MYD88 | NOD2 |
STAT1 | STAT3 | STAT4 | SYK | Tbet | TRAF3 | TRIF |
FURTHER INFORMATION
Margarida Saraivas homepage: http://www.icvs.uminho.
pt/icvs/domains/inf/cv/saraivam_cv_files/saraiva.m_cv.htm
Anne OGarras homepage: http://www.nimr.mrc.ac.uk/
immunoreg/ogarra/
All lInkS Are ACTIve In The onlIne pDf
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