Clostridium perfringens epsilon toxin mutant Y30A-Y196A as a recombinant vaccine candidate against enterotoxemia.
Epsilon toxin (Etx) is a β-pore-forming toxin produced by Clostridium perfringens toxinotypes B and D and plays a key role in the pathogenesis of enterotoxemia, a severe, often fatal disease of ruminants that causes significant economic losses to the farming industry worldwide. This study aimed to determine the potential of a site-directed mutant of Etx (Y30A-Y196A) to be exploited as a recombinant vaccine against enterotoxemia. Replacement of Y30 and Y196 with alanine generated a stable variant of Etx with significantly reduced cell binding and cytotoxic activities in MDCK.2 cells relative to wild type toxin (>430-fold increase in CT50) and Y30A-Y196A was inactive in mice after intraperitoneal administration of trypsin activated toxin at 1000× the expected LD50 dose of trypsin activated wild type toxin. Moreover, polyclonal antibody raised in rabbits against Y30A-Y196A provided protection against wild type toxin in an in vitro neutralisation assay. These data suggest that Y30A-Y196A mutant could form the basis of an improved recombinant vaccine against enterotoxemia.
College of Life and Environmental Sciences, University of Exeter, Stocker Road, Exeter EX4 4QD, United Kingdom. Electronic address: m.bokori-brown@exeter.ac.uk.
Monika
M
Bokori-Brown
College of Life and Environmental Sciences, University of Exeter, Stocker Road, Exeter EX4 4QD, United Kingdom.
Charlotte A
CA
Hall
College of Life and Environmental Sciences, University of Exeter, Stocker Road, Exeter EX4 4QD, United Kingdom.
Charlotte
C
Vance
College of Life and Environmental Sciences, University of Exeter, Stocker Road, Exeter EX4 4QD, United Kingdom.
Sérgio P
SP
Fernandes da Costa
Department of Biological Sciences, Birkbeck College, Malet Street, London WC1E 7HX, United Kingdom.
Christos G
CG
Savva
Department of Biological Sciences, Birkbeck College, Malet Street, London WC1E 7HX, United Kingdom.
Claire E
CE
Naylor
Department of Biological Sciences, Birkbeck College, Malet Street, London WC1E 7HX, United Kingdom.
Ambrose R
AR
Cole
Department of Biological Sciences, Birkbeck College, Malet Street, London WC1E 7HX, United Kingdom.
Ajit K
AK
Basak
Department of Biological Sciences, Birkbeck College, Malet Street, London WC1E 7HX, United Kingdom.
David S
DS
Moss
College of Life and Environmental Sciences, University of Exeter, Stocker Road, Exeter EX4 4QD, United Kingdom.
Richard W
RW
Titball
D000818:Animals; D001427:Bacterial Toxins / Q000276:immunology*; D001428:Bacterial Vaccines / Q000276:immunology*; D004285:Dogs; D004767:Enterotoxemia / Q000517:prevention & control*; D005260:Female; D061985:Madin Darby Canine Kidney Cells; D051379:Mice; D008807:Mice, Inbred BALB C; D016297:Mutagenesis, Site-Directed; D009500:Neutralization Tests; D017434:Protein Structure, Tertiary; D011817:Rabbits; D011994:Recombinant Proteins / Q000276:immunology; D014614:Vaccines, Synthetic / Q000276:immunology
D001427:Bacterial Toxins; D001428:Bacterial Vaccines; C039811:Clostridium perfringens epsilon-toxin; D011994:Recombinant Proteins; D014614:Vaccines, Synthetic
Clostridium perfringens; Enterotoxemia; Epsilon toxin pore-forming toxin; Recombinant vaccine