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Clostridium perfringens epsilon toxin vaccine candidate lacking toxicity to cells expressing myelin and lymphocyte

Helen Morcrette1, Monika Bokori-Brown1, Stephanie Ong1

  • 11University of Exeter, Exeter, EX4 4QD UK.

NPJ Vaccines
|August 3, 2019
PubMed

Insights

A modified Clostridium perfringens epsilon toxin (Y30A-Y196A-A168F) shows significantly reduced toxicity. This enhanced variant effectively induces protective antibodies, making it a promising candidate for a new enterotoxaemia vaccine.

Area of Science:

  • Veterinary Science
  • Microbiology
  • Toxicology

Background:

  • Clostridium perfringens epsilon toxin causes enterotoxaemia, a severe disease in livestock.
  • A previously developed variant (Y30A-Y196A) showed reduced toxicity but retained significant effects on certain cell types.
  • Understanding toxin-receptor interactions is crucial for developing effective vaccines.

Purpose of the Study:

  • To investigate the toxicity of epsilon toxin variants on cells expressing its receptor, myelin and lymphocyte protein (MAL).
  • To develop a more attenuated epsilon toxin variant with enhanced vaccine potential.
  • To evaluate the immunogenicity and protective capacity of the novel variant.

Main Methods:

  • Site-directed mutagenesis was used to create epsilon toxin variants, including Y30A-Y196A and Y30A-Y196A-A168F.
  • Toxicity assays were performed on Madin-Darby canine kidney (MDCK) cells, Chinese hamster ovary (CHO) cells expressing sheep MAL, and in vivo mouse models.
  • Immunization studies were conducted in rabbits and sheep, followed by antibody level assessments and challenge experiments.

Main Results:

  • The Y30A-Y196A variant unexpectedly showed no reduction in toxicity towards CHO cells expressing MAL.
  • The Y30A-Y196A-A168F variant demonstrated significantly reduced toxicity across all tested models: >3-fold reduction in MDCK cells, >4-fold in mice, and >200-fold in CHO-MAL cells.
  • Immunization with Y30A-Y196A-A168F in rabbits and sheep elicited robust and persistent neutralizing antibody responses against epsilon toxin for at least one year.

Conclusions:

  • The epsilon toxin variant Y30A-Y196A-A168F exhibits significantly attenuated toxicity compared to previous versions.
  • This highly attenuated variant successfully induces strong, long-lasting neutralizing antibodies.
  • Y30A-Y196A-A168F represents a promising candidate for the next generation of enterotoxaemia vaccines.
Keywords:
Bacterial infectionProtein vaccines

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