Production of neutralizing antibodies against the secreted Clostridium chauvoei toxin A (CctA) upon blackleg vaccination.
Animals
Antibodies, Bacterial
/ blood
Antibodies, Neutralizing
/ blood
Bacterial Toxins
/ immunology
Bacterial Vaccines
/ administration & dosage
Cattle
Cattle Diseases
/ prevention & control
Clostridium Infections
/ prevention & control
Clostridium chauvoei
/ immunology
Culture Media
/ chemistry
Enzyme-Linked Immunosorbent Assay
Guinea Pigs
Leukocidins
/ immunology
Neutralization Tests
Virulence Factors
/ immunology
Animal testing
Blackleg
CctA
Clostridium chauvoei
Haemolysin-neutralizing antibodies
Immunoblotting
Leucocidin
Mass spectrometry
Proteomics
Vaccination
Vaccine batch release tests
Journal
Anaerobe
ISSN: 1095-8274
Titre abrégé: Anaerobe
Pays: England
ID NLM: 9505216
Informations de publication
Date de publication:
Apr 2019
Apr 2019
Historique:
received:
12
10
2018
revised:
30
01
2019
accepted:
13
02
2019
pubmed:
17
2
2019
medline:
25
7
2019
entrez:
17
2
2019
Statut:
ppublish
Résumé
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.
Identifiants
pubmed: 30771460
pii: S1075-9964(19)30034-4
doi: 10.1016/j.anaerobe.2019.02.011
pii:
doi:
Substances chimiques
Antibodies, Bacterial
0
Antibodies, Neutralizing
0
Bacterial Toxins
0
Bacterial Vaccines
0
Culture Media
0
Leukocidins
0
Virulence Factors
0
Types de publication
Journal Article
Langues
eng
Pagination
78-87Informations de copyright
Copyright © 2019 The Authors. Published by Elsevier Ltd.. All rights reserved.