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Research Article Free access | 10.1172/JCI117080

Neutrophil recruitment in Clostridium difficile toxin A enteritis in the rabbit.

C P Kelly, S Becker, J K Linevsky, M A Joshi, J C O'Keane, B F Dickey, J T LaMont, and C Pothoulakis

Section of Gastroenterology, Boston University School of Medicine, Massachusetts 02118.

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Section of Gastroenterology, Boston University School of Medicine, Massachusetts 02118.

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Section of Gastroenterology, Boston University School of Medicine, Massachusetts 02118.

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Section of Gastroenterology, Boston University School of Medicine, Massachusetts 02118.

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Section of Gastroenterology, Boston University School of Medicine, Massachusetts 02118.

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Section of Gastroenterology, Boston University School of Medicine, Massachusetts 02118.

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Section of Gastroenterology, Boston University School of Medicine, Massachusetts 02118.

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Section of Gastroenterology, Boston University School of Medicine, Massachusetts 02118.

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Published March 1, 1994 - More info

Published in Volume 93, Issue 3 on March 1, 1994
J Clin Invest. 1994;93(3):1257–1265. https://doi.org/10.1172/JCI117080.
© 1994 The American Society for Clinical Investigation
Published March 1, 1994 - Version history
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Abstract

Neutrophil infiltration is a prominent feature of Clostridium difficile-associated enteritis and colitis. The aim of this study was to examine the importance of neutrophil recruitment and neutrophil-mediated tissue damage in C. difficile toxin A-induced enteritis. Competitive binding experiments using purified 3H-toxin A demonstrated the presence of a single class of medium affinity receptors on rabbit neutrophils (Kd 7 x 10(-8) M). Pertussis toxin and the nonhydrolyzable GTP analog GTPgamma S both inhibited 3H-toxin A binding (by 56 and 65%, respectively), indicating that the rabbit neutrophil toxin A receptor is G protein linked. Toxin A elicited a dose-dependent (25-200 micrograms/ml) stimulation of neutrophil migration in vitro, and this functional effect was also pertussis toxin sensitive (69% inhibition). Treatment of neutrophils with R15.7, a blocking monoclonal antibody to the leuocyte adhesion molecule CD18, inhibited toxin A-stimulated neutrophil migration by 85% in vitro. Pretreatment of rabbits with R15.7 also prevented neutrophil infiltration of toxin A-exposed ileal loops in vivo as determined by histologic examination and by ileal tissue myeloperoxidase levels. Furthermore, R15.7 effected a substantial inhibition of fluid secretion (by 65%), mannitol permeability (by 66%), and histologic damage in toxin A-exposed ileal loops. Anti-CD18 (R15.7) had no inhibitory effect on cholera toxin enterotoxicity. These data demonstrate that C. difficile toxin A is a proinflammatory toxin whose enterotoxic effects are substantially dependent upon neutrophil recruitment.

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