Differential ability of novel attenuated targeted deletion mutants ofFrancisella tularensissubspeciestularensisstrain SCHU S4 to protect mice against aerosol challenge with virulent bacteria: Effects of host background and route of immunization

Francisella tularensissubspeciestularensisis a highly virulent facultative intracellular pathogen of humans and a potential biological weapon. A live vaccine strain,F. tularensisLVS, was developed more than 50 years ago by pragmatic attenuation of a strain of the less virulentholarcticasubspecies. LVS was demonstrated to be highly effective in human volunteers who were exposed to intradermal challenge with fully virulent subsp.tularensis, but was less effective against aerosol exposure. LVS faces regulatory hurdles that to date have prevented its licensure for general use. Therefore, a better defined and more effective vaccine is being sought. To this end we have created gene deletion mutants in the virulent subsp.tularensisstrain and tested them for their ability to elicit a protective immune response against systemic or aerosol challenge with the highly virulent wild-type subsp. tularensis strain, SCHU S4. Both oral and intradermal (ID) primary vaccination routes were assessed in BALB/c and C3H/HeN mice as was oral boosting. One SCHU S4 mutant missing the heat shock gene,clpB, was significantly more attenuated than LVS whereas a double deletion mutant missing genesFTT0918andcapBwas as attenuated as LVS. In general mice immunized with SCHU S4ΔclpBwere significantly better protected against aerosol challenge than mice immunized with LVS. A single ID immunization of BALB/c mice with SCHU S4ΔclpBwas at least as effective as any other regimen examined. Mice immunized with SCHU S4Δ0918ΔcapBwere generally protected to a similar degree as mice immunized with LVS. A preliminary examination of immune responses to vaccination with LVS, SCHU S4ΔclpB,or SCHU S4Δ0918ΔcapBprovided no obvious correlate to their relative efficacies.