Immunogenicity Assessment of Clostridium Perfringens Type D Epsilon Toxin Epitope-based Chimeric Construct in Mice and Rabbit

Overview

Journal 3 Biotech

Publisher Springer

Specialty Biotechnology

Date 2020 Sep 1

PMID 32864287

Authors

Ajay Pratap Singh

Shyama N Prabhu

Viswas K Nagaleekar

Saroj K Dangi

Chandan Prakash

Vijendra Pal Singh

Affiliations

Soon will be listed here.

Abstract

Epsilon toxin (Etx) belongs to family of pore-forming toxin and is produced by 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 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 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).

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