An optimized, synthetic DNA vaccine encoding the toxin A and toxin B receptor binding domains of Clostridium difficile induces protective antibody responses in vivo (VAC7P.958)

Clostridium difficile -associated disease (CDAD) constitutes a large majority of nosocomial diarrhea cases within industrialized nations and is mediated by the effects of two secreted toxins, toxin A (TcdA) and toxin B (TcdB). Key toxin-neutralizing epitopes have been discovered within the carboxy-terminal receptor-binding domain (RBD) of the toxins, which has generated interest in developing the RBD as a vaccine target. While numerous platforms have been studied, very few data describe the potential of DNA vaccination against CDAD. Therefore, we created highly optimized plasmids encoding the RBD from TcdA and TcdB and immunized C57BL/6 mice and rhesus macaques intramuscularly followed by in vivo electroporation. In these animal models, vaccination induced significant levels of anti-RBD antibodies within the serum and feces that could neutralize toxins in an in vitro cytotoxicity assay. Moreover, mice that were either immunized with plasmids or given non-human primate immune sera were protected from a lethal intraperitoneal challenge of purified C. difficile toxins. Finally, immunized mice were significantly protected when challenged with strains of C. difficile spores that were homologous (VPI 10463; n=10/10) and heterologous (UK1; n=4/8) to our vaccine antigens. These data demonstrate the robust immunogenicity and efficacy of a TcdA/B RBD-based DNA vaccine in preclinical models of acute toxin-associated disease.