Beta toxin is essential for the intestinal virulence of Clostridium perfringens type C disease isolate CN3685 in a rabbit ileal loop model

Sameera Sayeed, Francisco A Uzal, Derek J. Fisher, Juliann Saputo, Jorge E. Vidal, Yue Chen, Phalguni Gupta, Julian I. Rood, Bruce A. McClane

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Abstract

Clostridium perfringens type C isolates, which cause enteritis necroticans in humans and enteritis and enterotoxaemias of domestic animals, typically produce (at minimum) beta toxin (CPB), alpha toxin (CPA) and perfringolysin O (PFO) during log-phase growth. To assist development of improved vaccines and therapeutics, we evaluated the contribution of these three toxins to the intestinal virulence of type C disease isolate CN3685. Similar to natural type C infection, log-phase vegetative cultures of wild-type CN3685 caused haemorrhagic necrotizing enteritis in rabbit ileal loops. When isogenic toxin null mutants were prepared using TargeTron® technology, even a double cpa/pfoA null mutant of CN3685 remained virulent in ileal loops. However, two independent cpb null mutants were completely attenuated for virulence in this animal model. Complementation of a cpb mutant restored its CPB production and intestinal virulence. Additionally, pre-incubation of wild-type CN3685 with a CPB-neutralizing monoclonal antibody rendered the strain avirulent for causing intestinal pathology. Finally, highly purified CPB reproduced the intestinal damage of wild-type CN3685 and that damage was prevented by pre-incubating purified CPB with a CPB monoclonal antibody. These results indicate that CPB is both required and sufficient for CN3685-induced enteric pathology, supporting a key role for this toxin in type C intestinal pathogenesis.

Original languageEnglish (US)
Pages (from-to)15-30
Number of pages16
JournalMolecular Microbiology
Volume67
Issue number1
DOIs
StatePublished - Jan 2008

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Clostridium perfringens
Enteritis
Virulence
Rabbits
Enterotoxemia
Monoclonal Antibodies
Pathology
Domestic Animals
Neutralizing Antibodies
Vaccines
Animal Models
Technology
Growth
Infection
Therapeutics

ASJC Scopus subject areas

  • Molecular Biology
  • Microbiology

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Beta toxin is essential for the intestinal virulence of Clostridium perfringens type C disease isolate CN3685 in a rabbit ileal loop model. / Sayeed, Sameera; Uzal, Francisco A; Fisher, Derek J.; Saputo, Juliann; Vidal, Jorge E.; Chen, Yue; Gupta, Phalguni; Rood, Julian I.; McClane, Bruce A.

In: Molecular Microbiology, Vol. 67, No. 1, 01.2008, p. 15-30.

Research output: Contribution to journalArticle

Sayeed, Sameera ; Uzal, Francisco A ; Fisher, Derek J. ; Saputo, Juliann ; Vidal, Jorge E. ; Chen, Yue ; Gupta, Phalguni ; Rood, Julian I. ; McClane, Bruce A. / Beta toxin is essential for the intestinal virulence of Clostridium perfringens type C disease isolate CN3685 in a rabbit ileal loop model. In: Molecular Microbiology. 2008 ; Vol. 67, No. 1. pp. 15-30.
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