Production of neutralizing antibodies against the secreted Clostridium chauvoei toxin A (CctA) upon blackleg vaccination.

Nicholson, Pamela; Furrer, Julia; Hässig, Michael; Strauss, Christian; Heller, Manfred; Braga, Sophie; Frey, Joachim (2019). Production of neutralizing antibodies against the secreted Clostridium chauvoei toxin A (CctA) upon blackleg vaccination. Anaerobe, 56, pp. 78-87. Elsevier 10.1016/j.anaerobe.2019.02.011

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Clostridium chauvoei is the etiologic agent of blackleg in cattle, inducing fever, severe myonecrosis, oedemic lesions and ultimately death of infected animals. The pathogen often results in such rapid death that antibiotic therapy is futile and thus vaccination is the only efficient strategy in order to control the disease. The β-barrel pore forming leucocidin Clostridium chauvoei toxin A (CctA) is one of the best characterised toxins of C. chauvoei and has been shown to be an important virulence factor. It has been reported to induce protective immunity and is conserved across C. chauvoei strains collected from diverse geographical locations for more than 50 years. The aim of this study was to identify the location of the CctA toxin during liquid culture fermentation and to use CctA to develop an in vitro assay to replace the current guinea pig challenge assay for vaccine potency in standard batch release procedures. We report that CctA is fully secreted in C. chauvoei culture and show that it is found abundantly in the supernatant of liquid cultures. Sera from cattle vaccinated with a commercial blackleg vaccine revealed strong haemolysin-neutralizing activity against recombinant CctA which reached titres of 1000 times 28 days post-vaccination. Similarly, guinea pig sera from an official potency control test reached titres of 600 times 14 days post-vaccination. In contrast, ELISA was not able to specifically measure anti-CctA antibodies in cattle serum due to strong cross-reactions with antibodies against other proteins present pre-vaccination. We conclude that haemolysin-neutralizing antibodies are a valuable measurement for protective immunity against blackleg and have the potential to be a suitable replacement of the guinea pig challenge potency test, which would forego the unnecessary challenge of laboratory animals.

Item Type:

Journal Article (Original Article)

Division/Institute:

05 Veterinary Medicine > Department of Infectious Diseases and Pathobiology (DIP) > Institute of Veterinary Bacteriology
04 Faculty of Medicine > Pre-clinic Human Medicine > BioMedical Research (DBMR)
04 Faculty of Medicine > Pre-clinic Human Medicine > BioMedical Research (DBMR) > DCR Services > Core Facility Massenspektrometrie- und Proteomics-Labor
04 Faculty of Medicine > Pre-clinic Human Medicine > BioMedical Research (DBMR) > Unit Childrens Hospital > Protein- und Zellbiologie

UniBE Contributor:

Nicholson, Pamela; Heller, Manfred and Braga, Sophie Marie-Pierre

Subjects:

600 Technology > 630 Agriculture
600 Technology > 610 Medicine & health

ISSN:

1095-8274

Publisher:

Elsevier

Language:

English

Submitter:

Marla Rittiner

Date Deposited:

27 Dec 2019 13:46

Last Modified:

27 Dec 2019 13:46

Publisher DOI:

10.1016/j.anaerobe.2019.02.011

PubMed ID:

30771460

Uncontrolled Keywords:

Animal testing Blackleg CctA Clostridium chauvoei Haemolysin-neutralizing antibodies Immunoblotting Leucocidin Mass spectrometry Proteomics Vaccination Vaccine batch release tests

BORIS DOI:

10.7892/boris.136304

URI:

https://boris.unibe.ch/id/eprint/136304

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