1887

Abstract

is a zoonotic, Gram-negative coccobacillus that causes tularemia in humans and animals. subspecies (type A) and subspecies (type B) are antigenically similar and more virulent than in humans. The genetic locus that encodes the LPS O antigen was found to be substantially different between the type B live vaccine strain (LVS) and One LVS-specific gene with homology to a galactosyl transferase was selected for allelic replacement using a –chloramphenicol expression suicide plasmid, and recombinants were screened for colony morphology on Congo red agar that matched that of . Two mutants (WbtI and WbtI) were isolated that contained substitutions in conserved motifs in the sugar transamine/perosamine synthetase (WbtI) of the O-antigen locus, and the latter mutant was extensively tested and characterized. WbtI grew at the same rate as the parent strain in Chamberlain's defined medium, completely lacked O antigen, was serum-sensitive but could grow in a mouse macrophage cell line, had increased resistance to sodium deoxycholate, and was highly attenuated in mice. Complementation of WbtI with the wild-type gene restored normal LPS synthesis, phenotypic properties similar to the parent, and virulence in mice. Immunization with WbtI protected mice against a relatively low-dose intraperitoneal challenge with LVS, but was less protective against a high-dose challenge. These results indicate that complete loss of O antigen alters the surface phenotype and abrogates virulence in , but also compromises the induction of full protective immunity against infection in mice.

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2007-09-01
2019-12-09
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