ABSTRACT: The immune system in the intestine must respond rapidly to invad-
ing pathogens without mounting sustained effector cell responses to the indig-
enous commensal bacteria. Results from this laboratory using the T cell
transfer model of colitis suggest that specialized populations of regulatory T
cells control the immune response in the intestine. Regulatory T (Tr) cell activ-
ity is enriched within the naturally arising CD4 CD25 Tr subset that has
been shown to prevent a number of inflammatory diseases. CD4 CD25 Tr
cells control intestinal inflammation induced by both innate and adaptive
immune responses via IL-10– and TGF-–dependent mechanisms. Recent
results have shown that CD4 CD25 Tr cells can cure established colitis,
suggesting their utility for the treatment of inflammatory bowel disease.
INTRODUCTION
The gastrointestinal (GI) tract is the largest interface between the immune system
and the outside world. It is home to a large number and diverse array of commensal
bacteria and is also one of the major surfaces through which pathogens gain access
to the body. Immune cells in the intestine are intimately associated with the epithe-
lium and are continually challenged by a myriad of antigens derived from food and
commensal bacteria, as well as pathogenic organisms. Specific anatomical features
of the intestine and the presence of specialized immune cells ensures rapid and local
responses to contain and eradicate pathogens without development of sustained
inflammatory responses to harmless intestinal antigens.1
The intestinal bacterial complex, composed of over 400 species in humans, exists
in an overall symbiotic relationship with the host, playing an important role in nutri-
tion, host defense, and organ development.2 Although the majority of commensal
bacteria are separated from immune cells by the epithelial cell barrier, there is good
Address for correspondence: Fiona Powrie, Sir William Dunn School of Pathology, University
of Oxford, South Parks Road, Oxford OX1 3RE, UK. Fax: +44-1865-275591.
fiona.powrie@path.ox.ac.uk
Ann. N.Y. Acad. Sci. 1029: 132–141 (2004). © 2004 New York Academy of Sciences.
doi: 10.1196/annals.1309.030
132
POWRIE: IMMUNE REGULATION IN THE INTESTINE 133
evidence that local immune responses are activated in response to intestinal bacte-
ria.3,4 Production of anti-inflammatory cytokines, such as IL-10 and TGF-β, is often
dominant during immune responses at mucosal surfaces, and this may be an impor-
tant adaptive mechanism to prevent the development of chronic inflammation.1
Inflammatory bowel disease (IBD), encompassing Crohn’s disease (CD) and
ulcerative colitis, is a chronic condition involving relapsing and remitting inflamma-
tion of the gastrointestinal tract.5 Clinical and experimental studies suggest that IBD
is the consequence of an aberrant inflammatory response to intestinal bacteria that
occurs in genetically susceptible individuals.6 Recently, mutations in the intracellu-
lar bacterial sensing protein, nucleotide-binding oligomerization domain 2 (NOD2),
have been shown to confer susceptibility to CD.7,8 Although it is not fully under-
stood how alterations in the NOD2 bacterial recognition pathway predispose to IBD,
the results do identify innate immune recognition pathways as important in control-
ling intestinal inflammation.
The complex multifactorial nature of IBD makes it difficult to dissect the role of
individual components in the etiology and pathogenesis of disease. Over the last
decade, a number of models of chronic intestinal inflammation have been described
that share features with human IBD (reviewed in Ref. 6). Results from these studies
have shown that IBD can develop as a consequence of altered intestinal barrier func-
tion, impaired innate immunity, excessive effector cell responses, or a deficiency in
regulatory pathways.
Studies from this laboratory using the T cell transfer model of colitis indicate that
CD4+ regulatory T cells (Tr) play a key role in intestinal homeostasis. Here, we
describe the properties of Tr cells that control intestinal inflammation and discuss
their application for therapy in IBD.
Dendritic cells (DCs) are abundant in the GI tract, in organized lymphoid struc-
tures, as well as in the lamina propria, where they may sample incoming antigens.
DCs express a number of pattern recognition receptors on their surface, allowing
them to detect and respond to the presence of microbes.18 Bacteria-induced activa-
tion of DCs leads to their maturation and migration to the secondary lymphoid
organs, where they are able to present antigen to naive T cells. In addition, mature,
activated DCs produce a number of inflammatory cytokines and costimulatory
molecules that are required for the development of sustained T cell responses.
In mice with colitis there is an accumulation of DCs in the mesenteric lymph
nodes (MLNs) as well as in colonic LP.19,20 DCs in the MLNs expressed an activated
phenotype with increased expression of CD40 and the TNF-like molecule CD134L.
Analysis of T cell proliferation in mice with colitis showed that up to 20 to 30% of
T cells were proliferating in the MLNs, consistent with the high proportion of acti-
vated DCs present.21 Activated T cells express the cell surface costimulatory mole-
cules CD40 ligand (CD40L) and CD134. CD40L binds CD40 on antigen-presenting
cells (APCs), inducing CD134L expression22 leading to the transmission of further
activatory signals to both the T cell and APC. The CD40-CD40L and CD134L-
CD134 pathways play functional roles in the inflammatory response, as blockade of
either pathway inhibits colitis.19,23,24
In addition to proliferating in the draining lymph nodes, colitogenic T cells also
proliferated locally in the colon. Proliferating T cells were typically located adjacent
to DCs in organized leukocyte clusters in the lamina propria.21 T cells were found
to home to DC clusters in the colon very early after transfer into immune-deficient
recipients, prior to the onset of colitis,20 making it tempting to speculate that colonic
leukocyte clusters also contribute to the initiation and perpetuation of the inflamma-
tory response in the intestine.
Together, the data suggest that interactions between activated DCs and activated
T cells drive intestinal inflammation, and that disruption of this positive-feedback
loop is sufficient to inhibit colitis.
An important theme that has emerged from the T cell transfer model is that func-
tionally specialized Tr cells capable of inhibiting intestinal inflammation are present
among the CD4+ T cell pool in normal mice. Initially identified as being contained
within the antigen-experienced CD4+ CD45RBlow population,11 Tr cell activity has
since been shown to be enriched within the CD4+ CD25+ subset.28 These “naturally
occurring” Tr cells inhibit T cell proliferation in vitro, and are important in the con-
trol of T cell responses to self-antigens, thus preventing the development of autoim-
mune disease.29 Recent studies have shown that CD4+ CD25+ Tr cells also play a
more general suppressive role, acting to limit immune pathology associated with
chronic immune stimulation.30
CD4+ CD25+ Tr cells account for 5 to 10% of peripheral T cells in mice, rats, and
humans.29 A feature that distinguishes CD4+ CD25+ Tr cells from other CD4+ T
cells is that they adopt their effector function in the thymus.29 Current evidence
suggests that CD4+ T cells bearing receptors with a high affinity for self-peptide will
differentiate into CD4+ CD25+ Tr cells following an encounter with cognate peptide
on thymic stromal cells.31 Foxp3, from the family of forkhead/winged helix tran-
scription factors, is differentially expressed by CD4+ CD25+ Tr cells and is required
for their development in the thymus.32,33 Retroviral transduction with Foxp3 leads
to the development of regulatory T cells from naive precursors, indicating that Foxp3
may be a master control switch for Tr cell development.34 Importantly, loss-of-func-
tion mutations in Foxp3 have been shown to be responsible for the lymphoprolifer-
ative disease that develops in scurfy mice, and for the human disease immune
polyendocrine enteropathy X-linked (IPEX) syndrome.35 Strikingly, patients with
IPEX develop a similar spectrum of diseases that develops in animal models in the
absence of regulatory T cells, including type 1 diabetes, allergy, and IBD-like
enteropathy.
the innate immune response but act to prevent chronic activation. Indeed, there is
evidence that stimulation via the toll-like receptor (TLR) pathway breaks Tr-
mediated control by a mechanism involving IL-6 production by activated DCs.37
The ability of CD4+ CD25+ Tr cells to control the innate immune response may
underlie the ability of these cells to inhibit chronic immune responses to a number
of infectious agents, and be an important host mechanism to prevent immune pathol-
ogy during persistent microbial infection.27,38,39
SPECIFICITY
The immune suppressive cytokines IL-10 and TGF-β play nonredundant roles in
intestinal homeostasis, as IL-10−/−45 and TGF-β1−/−46 mice develop chronic colitis.
POWRIE: IMMUNE REGULATION IN THE INTESTINE 137
phoid tissue and effector sites. Recently, a subset of CD4+ CD25+ Tr cells expressing
the integrin αE (CD103) was shown to home most efficiently to inflammatory sites
and to control acute inflammation. This suggests that distinct subsets of Tr cells may
control responses in lymphoid organs and tissues.58 Proliferation of CD4+ CD25+ Tr
cells was significantly reduced following resolution of the inflammatory response,
suggesting that CD4+ CD25+ Tr cells respond to the inflammation that they
regulate.21
In the MLN and colon, CD4+ CD25+ Tr cells localized at the interface between
pathogenic effector T cells and DCs and were in direct contact with both populations
of cells.21 This specific distribution in vivo may reflect the ability of activated DCs
to induce the activation and migration of CD4+ CD25+ Tr cells.59,60 Tr cells, in turn,
may suppress the immune response via effects on activated DCs1 or via direct effects
on effector T cells.62
OUTLOOK
The balance between effector and regulatory T cell responses is a key factor in
the development of intestinal inflammation. There is now a large body of evidence
showing that enhancement of Tr cell activity can prevent and even cure intestinal
inflammation in mouse models, suggesting novel therapeutic strategies for the treat-
ment of IBD. IPEX patients lacking functional Foxp3 protein develop severe enter-
opathy, suggesting that Tr cells also control intestinal inflammation in humans.
However, further studies are required to determine whether defects in Tr activity are
associated with genetic susceptibility to IBD in humans.
ACKNOWLEDGMENTS
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