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Francisella tularensis  is the causative agent of a fatal human disease, tularemia.  F. tularensis  was used in bioweapon programs in the past and is now classified as a category A select agent owing to its possible use in bioterror attacks. Despite over a century since its discovery, an effective vaccine is yet to be developed. In this study four transposon insertion mutants of  F. tularensis  live vaccine strain (LVS) in Na/H antiporter ( FTL_0304 ), aromatic amino acid transporter ( FTL_0291 ), outer membrane protein A (OmpA)-like family protein ( FTL_0325 ) and a conserved hypothetical membrane protein gene ( FTL_0057 ) were evaluated for their attenuation and protective efficacy against  F. tularensis  SchuS4 strain. All four mutants were 100–1000 fold attenuated for virulence in mice than parental  F. tularensis . Except for the  FTL_0304 , single intranasal immunization with the other three mutants provided 100% protection in BALB/c mice against intranasal challenge with virulent  F. tularensis  SchuS4. Differences in the protective ability of the  FTL_0325  and  FTL_0304  mutant which failed to provide protection against SchuS4 were investigated further. The results indicated that an early pro-inflammatory response and persistence in host tissues established a protective immunity against  F. tularensis  SchuS4 in the  FTL_0325  immunized mice. No differences were observed in the levels of serum IgG antibodies amongst the two vaccinated groups. Recall response studies demonstrated that splenocytes from the  FTL_0325  mutant immunized mice induced significantly higher levels of IFN-γ and IL-17 cytokines than the  FTL_0304  immunized counterparts indicating development of an effective memory response. Collectively, this study demonstrates that persistence of the vaccine strain together with its ability to induce an early pro-inflammatory innate immune response and strong memory responses can discriminate between successful and failed vaccinations against tularemia. This study describes a live attenuated vaccine which may prove to be an ideal vaccine candidate for prevention of respiratory tularemia.

Identification of a Live Attenuated Vaccine Candidate for Tularemia Prophylaxis