Direct evidence of mast cell involvement in Clostridium difficile toxin a—induced enteritis in mice

doi:10.1016/S0016-5085(98)70315-4

Gastroenterology

Volume 114, Issue 5, May 1998, Pages 956-964

https://doi.org/10.1016/S0016-5085(98)70315-4 Get rights and content

Abstract

Background & Aims: The pathogenesis of Clostridium difficile toxin A–induced intestinal inflammation is not completely understood. The aim of this study was to define the contribution of mast cells to the fluid secretion and neutrophil infiltration associated with toxin A–induced enteritis. Methods: Fluid secretion and neutrophil infiltration in toxin A– or buffer–challenged ileal loops were assessed in normal, mast cell–deficient, and mast cell–deficient Kit^W/Kit^W-v mice that had undergone selective repair of their mast cell deficiency. The effect of a specific substance P–receptor antagonist was also studied. Results: Intestinal fluid secretion and neutrophil recruitment were significantly diminished in mast cell–deficient Kit^W/Kit^W-v and mast cell–deficient Mgf^Sl/Mgf^Sl-d mice compared with the respective normal mice. Mast cell–reconstituted Kit^W/Kit^W-v mice showed responses similar to the normal congenic mice. Administration of a specific substance P–receptor antagonist (CP-96,345) reduced toxin A–induced intestinal fluid secretion and inhibited neutrophil infiltration in normal, mast cell–deficient Kit^W/Kit^W-v, and mast cell–reconstituted Kit^W/Kit^W-v mice. Conclusions: C. difficile toxin A elicits intestinal fluid secretion and neutrophil infiltration by both mast cell–dependent and –independent pathways, and substance P participates in both pathways.

GASTROENTEROLOGY 1998;114:956-964

Section snippets

Reagents

The specific substance P–receptor antagonist CP-96,345 {(2S,3S)-cis-2-(diphenylmetahyl)-N-[(2-methoxyphenyl)metahyl]-1-azabicyclo[2.2.2]octan-3-amine} and its inactive 2R,3R enantiomer CP-96,344 were provided by Pfizer Diagnostics. Toxin A was purified to homogeneity from culture supernatants of C. difficile strain 10,463 as previously described.²⁴

Animals

Male C57BL/6 mice (Charles River Laboratories, Wilmington, MA) and male genetically mast cell–deficient WBB6F₁-Kit^W/Kit^W-v mice (referred to as Kit^W

C. difficile toxin A–induced intestinal fluid secretion is diminished in mast cell–deficient Kit^W/Kit^W-v and Mgf^Sl/Mgf^Sl-d mice

We initially examined the ileal secretory response to different concentrations of toxin A in C57BL/6 mice (which are semisyngeneic with mast cell–deficient mice). We found that the injection of 10 μg of toxin A into ileal loops in C57BL/6 mice elicited a significant and reproducible secretory response (data not shown). Therefore, 10 μg of toxin A per ileal loop was used for the remainder of our studies.

We then compared the secretory responses in normal +/+ mice and mast cell–deficient Kit^W/Kit

Discussion

Our study shows that genetically mast cell–deficient mice have diminished secretory responses and neutrophil infiltration in response to C. difficile toxin A. The reduced response to toxin A in mast cell–deficient mice was found to be the result of their mast cell deficiency because selective, systemic reconstitution of mast cell populations in mast cell–deficient Kit^W/Kit^W-v mice restored intestinal fluid secretion and neutrophil infiltration to the levels found in normal (+/+) mice (Figure 3

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^☆: Address requests for reprints to: Barry K. Wershil, M.D., Combined Program in Pediatric Gastroenterology and Nutrition, Massachusetts General Hospital, 149 13th Street (1493404), Charlestown, Massachusetts 02129. e-mail: [email protected]; fax: (617) 667-3616.

^☆☆: Supported by National Institutes of Health grants DK33506 and Core B (Morphology), DK46819 and DK40561 (Clinical Nutrition Research Center at Harvard) (to B.K.W), and DK 47343 (to C.P.). Dr. I. Castagliulo was supported by a Research Fellowship from the Crohn's and Colitis Foundation of America, Inc.

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Alimentary TractDirect evidence of mast cell involvement in Clostridium difficile toxin a—induced enteritis in mice☆,☆☆

Abstract

Section snippets

Reagents

Animals

C. difficile toxin A–induced intestinal fluid secretion is diminished in mast cell–deficient KitW/KitW-v and MgfSl/MgfSl-d mice

Discussion

Gastroenterology

Gastroenterology

Gastroenterology

Gastroenterology

Gastroenterology

Gastroenterology

Gastroenterology

Cell Immunol

Biochem Pharmacol

J Allergy Clin Immunol

Gastroenterology

Blood

Blood

Immunity

Eur J Pharmacol

Gastroenterology

Gastroenterology

Clostridium difficile: its disease and toxins

Clin Microbiol Rev

Clostridium difficile colitis

N Engl J Med

Clostridium difficile toxin B is more potent than toxin A in damaging human colonic epithelium in vitro

J Clin Invest

Biological activities of toxins A and B of Clostridium difficile

Infect Immun

Effect of toxins A and B of Clostridium difficile on rabbit ileum and colon

Gut

Neutrophil recruitment in Clostridium difficile toxin A enteritis in the rabbit

J Clin Invest

Alimentary Tract
Direct evidence of mast cell involvement in Clostridium difficile toxin a—induced enteritis in mice☆,☆☆

C. difficile toxin A–induced intestinal fluid secretion is diminished in mast cell–deficient Kit^W/Kit^W-v and Mgf^Sl/Mgf^Sl-d mice