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Toxins 2015, 7(4), 1235-1252; doi:10.3390/toxins7041235

Binding Studies on Isolated Porcine Small Intestinal Mucosa and in vitro Toxicity Studies Reveal Lack of Effect of C. perfringens Beta-Toxin on the Porcine Intestinal Epithelium

1
Department of Infectious Diseases and Pathobiology, Institute of Animal Pathology, Vetsuisse Faculty, University of Bern, Bern 3012, Switzerland
2
Veterinary Public Health Institute, Vetsuisse Faculty, University of Bern, Bern 3012, Switzerland
3
Department of Pathology, Bacteriology and Avian Medicine, Ghent University, Ghent 9000, Belgium
*
Author to whom correspondence should be addressed.
Academic Editor: Teresa Krakauer
Received: 29 January 2015 / Revised: 18 March 2015 / Accepted: 31 March 2015 / Published: 9 April 2015
(This article belongs to the Special Issue Enterotoxins: Microbial Proteins and Host Cell Dysregulation)
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Abstract

Beta-toxin (CPB) is the essential virulence factor of C. perfringens type C causing necrotizing enteritis (NE) in different hosts. Using a pig infection model, we showed that CPB targets small intestinal endothelial cells. Its effect on the porcine intestinal epithelium, however, could not be adequately investigated by this approach. Using porcine neonatal jejunal explants and cryosections, we performed in situ binding studies with CPB. We confirmed binding of CPB to endothelial but could not detect binding to epithelial cells. In contrast, the intact epithelial layer inhibited CPB penetration into deeper intestinal layers. CPB failed to induce cytopathic effects in cultured polarized porcine intestinal epithelial cells (IPEC-J2) and primary jejunal epithelial cells. C. perfringens type C culture supernatants were toxic for cell cultures. This, however, was not inhibited by CPB neutralization. Our results show that, in the porcine small intestine, CPB primarily targets endothelial cells and does not bind to epithelial cells. An intact intestinal epithelial layer prevents CPB diffusion into underlying tissue and CPB alone does not cause direct damage to intestinal epithelial cells. Additional factors might be involved in the early epithelial damage which is needed for CPB diffusion towards its endothelial targets in the small intestine. View Full-Text
Keywords: Clostridium perfringens type C; beta-toxin; endothelium; mucosa; epithelium; pathogenesis; porcine Clostridium perfringens type C; beta-toxin; endothelium; mucosa; epithelium; pathogenesis; porcine
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Roos, S.; Wyder, M.; Candi, A.; Regenscheit, N.; Nathues, C.; van Immerseel, F.; Posthaus, H. Binding Studies on Isolated Porcine Small Intestinal Mucosa and in vitro Toxicity Studies Reveal Lack of Effect of C. perfringens Beta-Toxin on the Porcine Intestinal Epithelium. Toxins 2015, 7, 1235-1252.

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