Immunogenicity assessment of Clostridium perfringens type D epsilon toxin epitope-based chimeric construct in mice and r
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ORIGINAL ARTICLE
Immunogenicity assessment of Clostridium perfringens type D epsilon toxin epitope‑based chimeric construct in mice and rabbit Ajay Pratap Singh1 · Shyama N. Prabhu2 · Viswas K. Nagaleekar3 · Saroj K. Dangi3 · Chandan Prakash4 · Vijendra Pal Singh5 Received: 16 March 2020 / Accepted: 17 August 2020 © King Abdulaziz City for Science and Technology 2020
Abstract Epsilon toxin (Etx) belongs to family of pore-forming toxin and is produced by Clostridium perfringens type D. The Etx toxin is responsible for the pathogenesis of enterotoxaemia in sheep and goats, and occasionally in other livestock animals. The present study aimed to develop a Clostridium perfringens epsilon toxin-based chimeric epitope construct having immunodominant B-cell epitope and universal T-cell epitope and its immunogenicity was evaluated in mice and rabbit. An artificial chimeric epitope construct (CEC) was prepared by joining tandem repeats of a peptide containing amino acids (aa) 134–145 of epsilon toxin B-cell epitope and universal T-cell epitopes. The CEC was expressed in the Escherichia coli following codon optimization for efficient translational efficiency and purified by affinity chromatography. The antigenic reactivity of r-CEC proteins was confirmed by western blot with rabbit anti-r-Etox hyperimmune sera. The immunogenicity of the recombinant single CEC was examined in mice and rabbit by indirect ELISA. It was found that r-CEC yielded high titers of neutralizing antibodies (≥ 1.035 IU/ml) in immunized mice and rabbit. The potency of chimeric protein immunized serum was observed to be higher than the recommended level (0.1–0.3 IU/ml) for protection in sheep and goats. This indicated the potential ability of the chimeric protein as a vaccine candidate. This further requires studying the immune response in targeted host species (sheep and goat). Keywords Enterotoxaemia · Clostridium perfrienges · Epsilon toxin · Recombinant · B-cell epitope
1
Department of Veterinary Microbiology, College of Veterinary and Animal Science, COVSc.&AH, U.P. Pandit Deen Dayal Upadhyaya Pashu Chikitsa Vigyan Vishwavidyalaya Evam Go Anusandhan Sansthan (DUVASU), Mathura, UP 281001, India
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Saroj K. Dangi https://www.researchgate.net/scientific-contributions/204499 8288-Saroj-K-Dangi
Department of Veterinary Pathology, COVSc.&AH, U.P. Pandit Deen Dayal Upadhyaya Pashu Chikitsa Vigyan Vishwavidyalaya Evam Go Anusandhan Sansthan (DUVASU), Mathura, UP 281001, India
3
Chandan Prakash https://www.researchgate.net/scientific-contributions/205992 1277_C_Prakash
Division Bacteriology and Mycology, Indian Veterinary Reaserch Institute (IVRI), Izatnagar, Bareilly, Uttar Pradesh 243 122, India
4
Vijendra Pal Singh https://www.researchgate.net/scientific-contributions/508540 16-Vijendra-Pal-Singh
Centre for Advance Animal Research and Diagnosis, Indian Veterinary Reaserch Institute (IVRI), Izatnagar, Bareilly, Uttar Pradesh 243 122, India
5
National Institute of High Security Animal Disease (NISHAD), Bhopal, Madhya Pradesh
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