CD8+ Cytotoxic T Cells Induce Relapsing Colitis in Normal Mice

doi:10.1053/j.gastro.2006.05.018

Gastroenterology

Volume 131, Issue 2, August 2006, Pages 485-496

https://doi.org/10.1053/j.gastro.2006.05.018 Get rights and content

Background & Aims: Most mouse models of IBD have emphasized an effector role of type-1 CD4⁺ T cells in colitis. The aim of this study was to develop a model of antigen-specific relapsing colitis to investigate the relative contribution of CD4⁺ and CD8⁺ effectors.

Methods: Balb/C mice were sensitized and challenged with a suboptimal dose of 2.4 dinitrobenzene sulfonic acid to generate a colonic delayed-type hypersensitivity response. The respective role of CD4⁺ and CD8⁺ T cells in the initiation of colitis was analyzed by in vivo monoclonal antibody depletion and cell-transfer experiments. Dynamic and function of the colitogenic effectors were studied by immunohistochemistry, fluorescence-activated cell sorter analysis, enzyme-linked immunospot assay, quantitative polymerase chain reaction, and in vivo CTL assays.

Results: Relapsing colitis rapidly occurred only after challenge of previously sensitized mice. Interferon-γ–producing cytotoxic CD8⁺ T cells (Tc1) specific for hapten-modified self-proteins were generated in colon-draining lymph nodes on day 5 after sensitization, before the onset of disease. These CD8⁺ T cells were rapidly recruited upon challenge into colon lamina propria as granzyme B–expressing effectors exerting ex vivo cytotoxicity against syngeneic hapten-modified colonic epithelial cells. Colitis was prevented by in vivo antibody depletion of CD8⁺, but not of CD4⁺, T cells and could be induced in naive recipients within 48 hours after transfer of CD8⁺, but not CD4⁺, T cells purified from sensitized mice.

Conclusions: Our data show that antigen-specific CD8⁺ T cells can induce relapsing colitis in normal mice and suggest that the cytolytic function of CD8 Tc1 against epithelial cells may initiate the intestinal inflammatory process.

Section snippets

Mice

Balb/C male mice and syngeneic athymic nude male mice (6–10 weeks old) were purchased from Charles River laboratories (L’Arbresle, France). All experiments were previously approved by the Animal Care and Use Committee according to governmental guidelines and were performed in the accredited establishment, PBES of Ecole Normale Superieure de Lyon. All mice were fed with standard mice chow pellets ad libidum.

Model of DNBS-Specific T-Cell–Mediated Colitis

The model of hapten-specific colonic DTH was set up by using DNBS, a hapten comparable

Relapsing Colitis Results From a Hapten-Specific Colonic DTH Response

To induce DNBS-specific colonic DTH, Balb/C mice were immunized by intracolonic administration (enema) of a nontoxic dose of 2 mg of DNBS in 50% ethanol. This dose of hapten was determined in preliminary dose-response experiments as the minimal nontoxic dose of hapten unable to induce colitis or mortality in naive recipient (not shown). Five days later, colonic DTH was elicited by intracolonic challenge with 1 mg of DNBS. No sign of colitis or body weight loss was detected in day

Discussion

In this study, we developed and characterized a new model of antigen-specific colonic DTH response in normal mice, based on colonic sensitization and challenge with DNBS, and showed that specific IFN-γ–producing CD8⁺ cytotoxic T (Tc1) cells can induce acute and relapsing colitis.

The DNBS-specific DTH response was characterized by 2 temporally dissociated phases. The sensitization (ie, afferent) phase induced by intracolonic immunization with 2 mg of DNBS resulted within 5 days in the in vivo

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: Supported by an institutional grant from Institut National de la Santé et de la Recherche Médicale (INSERM) and grants from Institut de Recherche sur les Maladies de l’Appareil Digestif (IRMAD), from Académie Nationale de Médecine, and from Association François Aupetit.

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Basic–alimentary tractCD8+ Cytotoxic T Cells Induce Relapsing Colitis in Normal Mice

Section snippets

Mice

Model of DNBS-Specific T-Cell–Mediated Colitis

Relapsing Colitis Results From a Hapten-Specific Colonic DTH Response

Discussion

Immunity

Gastroenterology

Gastroenterology

Immunity

Gastroenterology

Gastroenterology

Blood

Gastroenterology

Gastroenterology

Immunity

Disparate CD4+ lamina propria (LP) lymphokine secretion profiles in inflammatory bowel disease. Crohn’s disease LP cells manifest increased secretion of IFN-gamma, whereas ulcerative colitis LP cells manifest increased secretion of IL-5

J Immunol

Inflammatory bowel disease

N Engl J Med

The immunological and genetic basis of inflammatory bowel disease

Nat Rev Immunol

Tolerance towards resident intestinal flora in mice is abrogated in experimental colitis and restored by treatment with interleukin-10 or antibodies to interleukin-12

Eur J Immunol

Phenotypically distinct subsets of CD4+ T cells induce or protect from chronic intestinal inflammation in C. B-17 scid mice

Int Immunol

Colitogenic Th1 cells are present in the antigen-experienced T cell pool in normal micecontrol by CD4+ regulatory T cells and IL-10

J Immunol

Th1-type responses mediate spontaneous ileitis in a novel murine model of Crohn’s disease

J Clin Invest

CD4+ T cells reactive to enteric bacterial antigens in spontaneously colitic C3H/HeJBir miceincreased T helper cell type 1 response and ability to transfer disease

J Exp Med

Characterization of the mucosal cell-mediated immune response in IL-2 knockout mice before and after the onset of colitis

Immunology

Inhibition of IFN-gamma-inducible protein-10 abrogates colitis in IL-10-/- mice

J Immunol

Basic–alimentary tract
CD8⁺ Cytotoxic T Cells Induce Relapsing Colitis in Normal Mice