A Francisella tularensis Live Vaccine Strain (LVS) Mutant with a Deletion in capB, Encoding a Putative Capsular Biosynthesis Protein, Is Significantly More Attenuated than LVS yet Induces Potent Protective Immunity in Mice against F. tularensis Challenge

Francisella tularensis, the causative agent of tularemia, is in the top category (category A) of potential agents of bioterrorism. The F. tularensis live vaccine strain (LVS) is the only vaccine currently available to protect against tularemia; however, this unlicensed vaccine is relatively toxic and provides incomplete protection against aerosolized F. tularensis, the most dangerous mode of transmission. Hence, a safer and more potent vaccine is needed. As a first step toward addressing this need, we have constructed and characterized an attenuated version of LVS, LVS Delta capB, both as a safer vaccine and as a vector for the expression of recombinant F. tularensis proteins. LVS Delta capB, with a targeted deletion in a putative capsule synthesis gene (capB), is antibiotic resistance marker free. LVS Delta capB retains the immunoprotective O antigen, is serum resistant, and is outgrown by parental LVS in human macrophage-like THP-1 cells in a competition assay. LVS Delta capB is significantly attenuated in mice; the 50% lethal dose (LD50) intranasally (i.n.) is > 10,000-fold that of LVS. Providing CapB in trans to LVS Delta capB partially restores its virulence in mice. Mice immunized with LVS Delta capB i.n. or intradermally (i.d.) developed humoral and cellular immune responses comparable to those of mice immunized with LVS, and when challenged 4 or 8 weeks later with a lethal dose of LVS i.n., they were 100% protected from illness and death and had significantly lower levels (3 to 5 logs) of LVS in the lung, liver, and spleen than sham-immunized mice. Most importantly, mice immunized with LVS Delta capB i.n. or i.d. and then challenged 6 weeks later by aerosol with 10x the LD50 of the highly virulent type A F. tularensis strain SchuS4 were significantly protected (100% survival after i.n. immunization). These results show that LVS Delta capB is significantly safer than LVS and yet provides potent protective immunity against virulent F. tularensis SchuS4 challenge.