The
clostridial neurotoxin (CNT) family includes
botulinum neurotoxin (BoNT), serotypes A, B, E, and F of which can cause human
botulism, and
tetanus neurotoxin (TeNT), which is the causative agent of
tetanus. This suggests that the greatest need is for a multivalent or multiagent
vaccine that provides protection against all 5 agents. In this study, we investigated the feasibility of generating several pentavalent replicon
vaccines that protected mice against BoNTs and TeNT. First, we evaluated the potency of individual replicon
DNA or particle
vaccine against TeNT, which induced strong antibody and protective responses in BALB/c mice following 2 or 3 immunizations. Then, the individual replicon TeNT
vaccines were combined with tetravalent BoNTs
vaccines to prepare 4 types of pentavalent replicon
vaccines. These replicon
DNA or particle
pentavalent vaccines could simultaneously and effectively induce antibody responses and protect effects against the 5 agents. Finally, a solid-phase assay showed that the sera of pentavalent replicon formulations-immunized mice inhibited the binding of
THc to the
ganglioside GT1b as the sera of individual replicon
DNA or particle-immunized mice. These results indicated these pentavalent replicon
vaccines could protect against the 4 BoNT serotypes and effectively neutralize and protect the TeNT. Therefore, our studies demonstrate the utility of combining replicon
DNA or particle
vaccines into multi-agent formulations as potent
pentavalent vaccines for eliciting protective responses against BoNTs and TeNT.