DOI 10.1007/s00535-007-2111-3
Review
Inflammatory bowel disease, past, present and future: lessons from
animal models
ATSUSHI MIZOGUCHI1,2 and EMIKO MIZOGUCHI2,3
1
Department of Pathology, Experimental Pathology, Simches 8234, Massachusetts General Hospital, 185 Cambridge Street, Boston, MA,
02114 USA
2
Center for the Study of Inflammatory Bowel Disease, Massachusetts General Hospital and Harvard Medical School, Boston, MA, USA
3
Department of Medicine, Massachusetts General Hospital and Harvard Medical School, Boston, MA, USA
Introduction
Inflammatory bowel disease (IBD), comprising Crohns
disease (CD) and ulcerative colitis (UC), is a chronic
inflammatory disorder caused by multifactorial conditions in a genetically predisposed host.13 The complicated mechanism underlying IBD is clearly indicated by
the presence of many IBD susceptibility genes.4 Three
Th17/IL-23
Accumulating data obtained from experimental IBD
models have suggested the involvement of both common
and distinct mechanisms in the pathogenesis of UC
versus CD. One of the major concepts previously used
to explain the different mechanisms is the T helper (Th)
1/Th2 paradigm.811,13 Indeed, Th2 cytokines such as IL-4
and IL-13 contribute significantly to chronic UC-like
T H1
Mdr1a
CBI-b
Smad3
PAST
TNF
(ARE)
Common
IL-10
Gi2
IL-2R
IL-2
IL-2R
CD3
TGF
IL-15
IBD
HLAB27
IL-7
gp39
CdCS
IL-4
NCAD
GFAP-HA
Tir8
Tn
A20
TH1
LIGHT
Muc2
WASP
Keratin8
STAT4
Runx2
OT
R
E
H
TH2
Fig. 1. Deficiency (knockout mice, red colors) or overexpression (transgenic mice, blue colors) of many different molecules
in mice has been shown to induce the spontaneous development of colitis, which is mediated preferentially by an acquired
(ellipses) or innate (rectangles) immune pathway. The pathogenesis of colitis in some mouse models has significant contributions from the T helper (Th) 1 or Th2 pathway. IBD,
inflammatory bowel disease; IL, interleukin; TGF, transforming growth factor; A20, also known as tumor necrosis factor
(TNF)-induced protein 3; CBl-b, E3 ubiquitin ligase; Gi2, G
protein i2; GFAP-HA, transgenic mice in which enteric glia
are specifically disrupted; GPX, glutathione peroxidase;
HLAB27, HLAB27/human 2 microglobulin transgenic rats;
LIGHT, a TNF superfamily member; Mdr, multiple drug resistance; Muc2, mucin2; NCAD, transgenic mice that overexpress
dominant negative N-cadherin in the intestinal epithelial cells;
NEMO, NF-B essential modulator; NFAT, nuclear factor of
activated T cells; STAT, signal transducer and activator of
transcription; TCR, T-cell receptor; Tir8, also known as
SIGIRR (single Ig interleukin-1-related receptor); TNFARE,
mice lacking TNF AU-rich elements; WASP, Wiskott-Aldrich
syndrome protein; XBP1, X-box binding protein
disease.811 In contrast, the Th1 pathway has been implicated in the pathogenesis of CD.811,13 In addition to the
Th1/Th2 theory, accumulating recent studies have
unveiled the critical involvement of the IL-23/IL-17
pathway in the pathogenesis of CD-like experimental
diseases.1418 IL-12 (p40/p35) has generally been believed
to be a crucial factor involved in the development of
Th1-mediated colitis because of the beneficial effect of
anti-IL-12p40 treatment in CD patients as well as in
Th1-mediated experimental colitis.11 However, recent
studies have demonstrated that IL-23 (a heterodimer of
p40 and p19 subunits) rather than IL-12 (a heterodimer
of p40 and p35 subunits) contributes to the development of Th1-mediated chronic colitis observed in several
murine CD models, including IL-10 KO mice,14 the
CD45RB model,16,17 the cecal bacteria-specific T cellinduced colitis model,18 the Helicobacter hepaticus-
TH2
IL-4
Treg
RA
Tn TGF
IL-12/IFN
NEMO
TCR
TH2
PRESENT
IL-12/IFN
TH1
IL-6
TH17
IL-23
the colon of CD patients, regardless of disease severity.30,31 In contrast, IL-17 expression becomes detectable
only in the moderately to severely affected colon of UC
patients, and the expression levels of colonic IL-17 are
significantly lower in UC than in CD.30,31 The pathogenic
role of IL-17 has been demonstrated in some colitis
models. Neutralization of IL-17A activity results in the
suppression of Th1-mediated chronic colitis in IL-10 KO
mice,14 and the absence of IL-17 receptor in mice induces
resistance to the Th1-mediated acute colitis induced by
TNBS administration.32 In addition, activation of an
IL-17 receptor-mediated signaling pathway via Act1 in
epithelial cells contributes to the development of DSSinduced intestinal injury.33 In contrast, neutralization of
systemic IL-17A activity by administration of anti-IL17A monoclonal antibodies has been shown previously
to exacerbate DSS-induced intestinal injury.34 IL-17A
does not seem to be involved in Th2-mediated chronic
colitis, which develops spontaneously in TCR KO
mice.35 The regulatory role of IL-17 has been documented in a Th2-mediated asthma model.36 Therefore,
the role of IL-17 in the pathogenesis of colitis may differ
distinctly, depending upon, for example, whether the
phase of colitis is chronic or acute, whether Th1 or Th2
has contributed to the disease, and whether IL-17 activity is systemic or local. Interestingly, a recent study has
shown that Treg cells, which are well known to suppress
Th1-mediated colitis in the CD45RB model, are incapable of suppressing IL-17-mediated inflammation.37
Finally, it is important to mention a very recent observation that a T-cell subset producing both IL-17 and IFN-
is present in the inflamed intestine but not in the peripheral blood of CD patients.38
IL-22, which belongs to the IL-10 cytokine family,
specifically targets innate, but not acquired, immune
responses through the activation of STAT3.39 Interestingly, similar to the expression pattern of IL-17, colonic
IL-22 expression is specifically induced by inflammation
in both CD and UC, but the expression level of IL-22 is
significantly higher in CD than in UC.4042 Indeed, recent
studies have demonstrated that, in addition to IL-17,
IL-22 is also expressed preferentially by the Th17 cell
subset.43,44 Functionally, IL-22 has recently been shown
to mediate IL-23-induced inflammation in the skin.44 In
contrast, our recent study demonstrated that IL-22 contributes to the amelioration of ongoing Th2-mediated
chronic colitis, in which IL-23 is not upregulated (unpublished data). Since IL-22 possesses the ability to stimulate production of both regulatory (e.g., IL-10,
antibacterial peptide, and SOCS3) and deleterious (e.g.,
IL-8) molecules,3941,45 the reciprocal regulatory versus
deleterious functions of IL-22 may be determined by
the disease dependence on IL-23.
Pathway 1
Bacteria
Pathway 3
Pathway 2
Ags
IC
Ags
M cells
FcRn
DCs
T cells
IgG
APC
Fig. 3. There are at least three pathways involved in the transportation of luminal antigens (Ags) to the intestinal lamina propria
(LP). M cells capture luminal Ags and then present them to T cells (Pathway 1). LP dendritic cells (DCs) directly capture the
luminal antigens by extending their dendrites to the intestinal lumen across epithelial cells (Pathway 2). Neonatal Fc receptor
(FcRn) serves as the vehicle that transports LP immunoglobulin G (IgG) across the colonic epithelial layer into the lumen where
the IgG can bind enteric bacterial antigens to form immune complexes (IC). The FcRn then recycles the antigens/IgG IC back
across the colonic epithelial cells into the LP for processing by antigen-presenting cells (APCs) such as DCs, which are capable
of presenting the antigens to T cells (Pathway 3)
PAST
PRESENT
B cells
B cells
2nd APCs
IL-12
Igs
Igs
IL-10
Fig. 4. In the past, immunoglobulin (Ig) production had generally been believed as the major function of B cells. However,
recent accumulating data have unveiled additional novel functions of B cells. B cells possess the abilities to produce several
cytokines such as IL-10 and IL-12 under inflammatory conditions and to serve as a secondary APCs to dampen T-cell
activation
healthy individuals or by LPS-stimulated B cells. Therefore, macrophages and DCs but not B cells have generally been believed to produce IL-12. However, this
concept may need to be revised in light of recent accumulating evidence that production of IL-12 by B cells
becomes easily detectable under certain inflammatory
conditions and when unmethylated CpG dinucleotides
(CpG), which became available for use only recently,
but not LPS, are used for the stimulation.77,85 Indeed,
we recently found that a Th2-mediated intestinal inflammatory condition as well as CpG stimulation induces
the differentiation of IL-12-producing B cells in the
presence of IL-10.35 Importantly, the inducible IL-12producing B cells contributed to the amelioration of
this Th2-mediated colitis. This finding may support
the earlier observations that Toll-like receptor (TLR)
9/CpG signaling contributes to the suppression of
colitis.8688 Interestingly, absence of IL-12-producing B
cells enhances the production of IL-17A by CD4+ T
cells.35 Indeed, IL-12 has previously been demonstrated
to inhibit IL-17 production.89 These data indicate that B
cells possess considerably more functional plasticity
than previously believed (Fig. 4).
The regulatory role of B cells in colitis and ileitis has
been demonstrated by several studies using different
IBD models, including TCR KO mice, Gi2 KO mice,
NFATc KO mice, and the CD45RB model.71,9096 Wei
and colleagues94 have identified a critical cellular
network for B cell-mediated regulation of colitis. MLN
B cells suppress the chronic colitis of Gi2 KO mice by
cooperating with regulatory CD8+ T cells and natural
killer T cells.94 In addition, recent studies in other autoimmune disease models have demonstrated that B cells
indirectly or directly enhance Treg cell function.9799 The
B cells that possess regulatory functions independent of
immunoglobulin production have been termed Breg
cells.95 It is important to mention that B cells, which
NOD2 (CARD15)
One of the significant advances in IBD studies in the
last decade is the discovery of nucleotide-binding oligomerization domain 2 (NOD2)/caspase-recruitment
domain 15 (CARD15) as a CD susceptibility gene by
Hugot et al.103 and Ogura et al.104 Since recent advances
in IBD genetics and NOD2 biology have already been
clearly introduced in excellent reviews,1,3,4,103106 this
review focuses only on findings in murine models regarding the role of NOD2 in the pathogenesis of IBD.
NOD2 induces innate immune responses through
cytosolic recognition of bacterial components such as
muramyl dipeptide (MDP), which is a conserved structure in virtually all types of peptidoglycans (PGN).104 To
explore the role of NOD2 mutation in the pathogenesis
of CD, three independent research groups have generated NOD2 KO mice107,108 or NOD2939ic knockin (KI)
mice, which carry a NOD2 gene mutated at position
2939 (corresponding to 3020 in human NOD2).109 In the
NOD2 KO mice generated by Kobayashi et al.,108 protective innate immune responses mediated by NOD2
recognition of MDP are completely abolished. Although
the NOD2 KO mice do not spontaneously develop
colitis, they are more susceptible, with a significantly
reduced production of an antibacterial peptide (cryptidin) to Listeria monocytogenesis infection but not to
DSS-induced intestinal injury.108 Abnormally low antibacterial peptide production in the ileum (mainly by
Paneth cells) is likely a primary defect in CD.110 Watanabe and colleagues111,112 proposed a role of NOD2 in
the suppression of the TLR2-dependent innate immune
pathway involved in the activation of the Th1 response
in studies using the other NOD2 KO mice generated by
Pauleau and Murray.107 The NOD2 deficiency significantly enhances the production of IFN-, IL-18, and
IL-12p70 by the splenocytes in response to stimulation
with PGN111 and increases the susceptibility of mice to
TLR2-dependent colitis induced by ovalbumin (OVA)specific T cells in the presence of OVA-producing Escherichia coli (ECOVA).112 In contrast, alteration of such
TLR2-mediated immune responses is not observed in
TLRs
The identification of mammalian TLRs as pattern recognition receptors for specific recognition of bacterial
products by Medzhitov et al.120 and Hoshino et al.121
led to major breakthroughs in many research fields,
including IBD studies. Recent advances in TLR biology
are well summarized in excellent reviews by the pioneers.122124 Therefore, here we focus only on the functional role of TLRs in the pathogenesis of colitis as
revealed by data accumulated during the last 5 years or
so.
TLR2 and TLR4
The role of TLR4 and TLR2 and their common adaptor
protein, myeloid differentiation factor 88 (MyD88), in
Future topics
Basic science has been continuously progressive both
technically and conceptually. In the following section,
we introduce attractive novel avenues of investigation
in IBD research and discuss classical, but clinically relevant, subjects that might contribute to the immediate
translation from bench to bedside of concepts for
improving the lives of patients with IBD.
Stem cells
A major current topic in science and medicine is stem
cell biology. Further advances in this field are guaranteed to contribute significantly to breakthroughs in the
development of novel therapeutic strategies for several
diseases. Importantly, the relevance of stem cell biology
10
11
Other topics
References
Conclusion
Finding accumulated in the past and more recently
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for developing novel therapeutic strategies to save the
lives of patients with IBD. Since the roles of several
molecules in the pathogenesis of IBD distinctly differ
depending on the type of cell, therapeutic strategies that
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