Macrophage-derived IL-18–mediated intestinal inflammation in the murine model of Crohn's disease

doi:10.1053/gast.2001.28021

Gastroenterology

Volume 121, Issue 4, October 2001, Pages 875-888

https://doi.org/10.1053/gast.2001.28021 Get rights and content

Abstract

Background & Aims: Crohn's disease (CD) is associated with an increased number of infiltrating macrophages, which release a variety of proinflammatory cytokines. Interleukin (IL)-18 has been implicated in the modulation of mucosal CD4⁺ T cells towards Th1 responses, which are implicated in the pathogenesis of CD. Here we assess the role of macrophages and of IL-18 in the murine model of intestinal inflammation that mimics the immunologic characteristics of human CD. Methods: Colitis was induced in C57BL/6 mice immunized with 2,4,6-trinitrobenzene sulfonic acid (TNBS) followed by rectal administration of TNBS in ethanol. Mice were treated with either an antibody directed against macrophages conjugated to the ribosome-inactivating protein saporin (anti–Mac-1-saporin) or with a neutralizing antibody against IL-18. In addition, we assessed whether an identical TNBS immunization/challenge protocol could induce colitis in IL-18^−/− mice. Results: The colonic mucosa of TNBS-treated mice was marked by infiltration of Mac-1–positive macrophages and up-regulation of IL-18. The administration of the anti–Mac-1-saporin antibody or the neutralizing anti–IL-18 antibody resulted in a dramatic attenuation of mucosal inflammation in this model. In addition, TNBS was unable to induce significant colitis in the IL-18^−/− mice. Conclusions: Our data underscore the pivotal role of macrophages, and the macrophage-derived IL-18, in the establishment of TNBS-induced colitis in mice. Our results highlight the potential use of therapy directed against IL-18 in the treatment of patients with CD.

GASTROENTEROLOGY 2001;121:875-888

Section snippets

Mice

Normal (IL-18^+/+) C57BL/6J mice were purchased from Japan Clea (Tokyo, Japan). IL-18–deficient mice C57BL/6J (IL-18^−/−) were generated as described previously³⁰ and maintained in the Animal Care facility of Keio University.

Cytokines and antibodies

A rat monoclonal antibody (mAb) to murine Mac-1 (CD11b) conjugated to saporin (anti–Mac-1-saporin, or MAC-1-ZAP) was obtained from Advanced Targeting Systems (Carlsbad, CA). The neutralizing anti–IL-18 antibody was prepared from sera of rabbits immunized with murine

Wasting disease in C57BL/6J mice by immunization of TNBS/BSA followed by intrarectal administration of TNBS/EtOH

Based on the previous report showing that TNBS immunization followed by rectal administration of TNBS in EtOH induces profound colitis in BALB/c mice,¹⁰ we explored the possibility that this protocol could similarly induce a chronic inflammation in the colon of C57BL/6J mice. In the current study, we found that systemic immunization of TNBS conjugated with BSA followed by intrarectal administration of TNBS/EtOH reproducibly developed pancolitis in almost all treated C57BL/6J mice. Specifically,

Discussion

The results of the present study clearly show the crucial role of infiltrating macrophages, and, more specifically, the proinflammatory cytokine IL-18 derived from activated macrophages, in the pathogenesis of experimental colitis induced by the hapten TNBS. These conclusions were supported by the demonstration that antibody treatment that targeted the effector macrophage or IL-18 itself dramatically prevented the establishment of TNBS-induced colitis. The results were further strengthened by

Acknowledgements

The authors thank Dr. Shigeo Koyasu for helpful discussion, Drs. Yasushi Iwao and Naoya Sakamoto for technical assistance, and Reiko Fujisaki for manuscript preparation.

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^☆: Address requests for reprints to: Toshifumi Hibi, M.D., Keio Cancer Center, 35 Shinanomachi, Shinjuku-ku, Tokyo 160-8582, Japan. e-mail: [email protected]; fax: (81) 3-3357-6156.

^☆☆: Supported in part by grants-in-aid from the Japanese Ministry of Education, Culture and Science, the Japanese Ministry of Health and Welfare, Chiyoda Mutual Life Foundation, Japan Health Sciences Foundation, and Keio University Medical Science Fund, Tokyo, Japan.

View full text

Alimentary TractMacrophage-derived IL-18–mediated intestinal inflammation in the murine model of Crohn's disease☆,☆☆

Abstract

Section snippets

Mice

Cytokines and antibodies

Wasting disease in C57BL/6J mice by immunization of TNBS/BSA followed by intrarectal administration of TNBS/EtOH

Discussion

Acknowledgements

Gastroenterology

Curr Opin Immunol

Gastroenterology

Gastroenterology

Clin Immunol Immunopathol

Methods

Curr Opin Immunol

J Biol Chem

Curr Opin Immunol

Gastroenterology

Immunity

J Immunol Methods

Blood

Immunity

Inflammatory bowel disease (first of two)

N Engl J Med

Immunological diseases of the gastrointestinal tract

Role of the intestinal microflora in pathogenesis and complications

Dendritic cells and scavenger macrophages in chronic inflammatory bowel disease

Gut

Changes in phenotypically distinct mucosal macrophage populations may be prerequisite for the development of inflammatory bowel disease

Clin Exp Immunol

Mucosal immunoregulation and inflammatory bowel disease: new insights from murine models of inflammation

Scand J Immunol

A synthetic mimetic of CD4 is able to suppress disease in a rodent model of immune colitis

Eur J Immunol

Antibodies to interleukin 12 abrogate established experimental colitis in mice

J Exp Med

Up-regulation of the IL-12/STAT-4 signaling pathway distinguishes Crohn's disease from ulcerative colitis (abstr)

Gastroenterology

Alimentary Tract
Macrophage-derived IL-18–mediated intestinal inflammation in the murine model of Crohn's disease☆,☆☆