Cytotoxin CctA, a major virulence factor of Clostridium chauvoei conferring protective immunity against myonecrosis.

Frey, Joachim; Johansson, Anders; Bürki, Sibylle; Vilei, Edy; Redhead, Keith (2012). Cytotoxin CctA, a major virulence factor of Clostridium chauvoei conferring protective immunity against myonecrosis. Vaccine, 30(37), pp. 5500-5505. Elsevier 10.1016/j.vaccine.2012.06.050

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OBJECTIVE The purpose of this study was to determine the identity of the major toxin of Clostridium chauvoei, an important pathogen of cattle causing black leg and to determine its value as a protective antigen in vaccines against myonecrosis. METHODS Genomic sequence analysis was used to determine potential virulence genes of C. chauvoei. Subsequently, the putative toxin candidate gene was cloned and expressed to obtain recombinant toxin. This toxin was investigated for its cytotoxic activity, hemolysis and its potential as a protective antigen in the guinea pig potency assay. RESULTS A novel protein toxin, named Clostridium chauvoei toxin A (CctA) that belongs to the family of β-barrel pore forming toxins of the leucocidin superfamily of bacterial toxins was discovered by whole genome sequence analysis. The corresponding gene cctA was found in all strains of C. chauvoei analyzed, isolated from various geographical areas over the globe during the last 50 years, but not in other pathogenic Clostridium species. Native CctA and recombinant rCctA produced in Escherichia coli in the form of a rCctA::NusA fusion protein or thrombin processed rCctA were highly cytotoxic for Embryonic Calf Nasal Epithelial (ECaNEp) cells and had high haemolytic activity against sheep erythrocytes in standard haemolysis assays. Polyclonal anti-rCctA rabbit antibodies fully neutralized the cytotoxic and haemolytic activity, not only of rCctA but also of supernatants from cultures of the various C. chauvoei strains, indicating that CctA is the main cytotoxic and haemolytic substance secreted by C. chauvoei. Using a standard vaccine release procedure, we demonstrated that vaccination of guinea pigs with CctA in the form of a fusion protein with the E. coli heat labile toxin B subunit (rCctA::LTB) as a peptide adjuvant protected the animals against challenge with spores of virulent C. chauvoei. CONCLUSIONS CctA is the major virulence factor of C. chauvoei and the main protective antigen in vaccines against blackleg.

Item Type:

Journal Article (Original Article)

Division/Institute:

05 Veterinary Medicine > Research Foci > Host-Pathogen Interaction
05 Veterinary Medicine > Department of Infectious Diseases and Pathobiology (DIP)
05 Veterinary Medicine > Department of Infectious Diseases and Pathobiology (DIP) > Institute of Veterinary Bacteriology

UniBE Contributor:

Frey, Joachim; Johansson, Anders; Bürki, Sibylle and Vilei, Edy

Subjects:

600 Technology > 630 Agriculture

ISSN:

0264-410X

Publisher:

Elsevier

Language:

English

Submitter:

Monika Mumenthaler

Date Deposited:

21 Aug 2018 12:51

Last Modified:

21 Aug 2018 12:51

Publisher DOI:

10.1016/j.vaccine.2012.06.050

PubMed ID:

22749595

BORIS DOI:

10.7892/boris.119392

URI:

https://boris.unibe.ch/id/eprint/119392

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