Construction, Characterization, and Animal Testing of WRSd1, a Shigella dysenteriae1 Vaccine

ABSTRACTWRSd1 is a Shigella dysenteriae1 vaccine containing deletions of the virG(icsA)gene required for intercellular spreading and a 20-kb chromosomal region encompassing the Shiga toxin genes (stxAB). WRSd1 was constructed from S. dysenteriae1 strain 1617 that was originally isolated during the 1968 to 1969 epidemic of Shiga dysentery in Guatemala. The virG(icsA)deletion was constructed from a streptomycin-resistant (Strr) mutant of 1617 by a filter mating procedures using a virG(icsA)deletion derivative, pΔvirG2. A colony that was invasive for HeLa cells and negative for the virG(icsA)gene by Southern blotting was grown anaerobically on plates containing chlorate for selection of resistant colonies that had lost the entire Shiga toxin gene. A virG(icsA) stxABStrrmutant selected from the chlorate plates was designated WRSd1. This candidate vaccine was evaluated for safety, immunogenicity, and protective efficacy using the guinea pig keratoconjunctivitis model. WRSd1 was Sereny negative, and two applications of this strain to the cornea elicited a significant protective immune response against the S. dysenteriae1 O antigen. Vaccination with WRSd1 conferred protection against challenge with each of three virulent S. dysenteriae1 strains. Since a vaccine protecting against multiple Shigellaspecies is required for most areas where Shigellais endemic, protection studies using a combination vaccine of Shigella sonneivaccine strain WRSS1, Shigella flexneri2a vaccine strain SC602, and WRSd1 were also performed. Guinea pigs vaccinated with a mixture of equal amounts of the three vaccine strains were protected against challenge with each of the homologous virulent strains. Unlike WRSS1 and SC602, however, the level of protection afforded by WRSd1 in a combination vaccine was lower than the protection elicited by a pure culture. A current Good Manufacturing Practice product of WRSd1 given intragastrically to rhesus monkeys proved safe and immunogenic.