Genetic Modification of the O-Polysaccharide of Francisella tularensis Results in an Avirulent Live Attenuated Vaccine

Abstract

Francisella tularensis, the causative agent of tularemia, is a highly virulent microbe. One significant virulence factor of F. tularensis is the O-polysaccharide (O-PS) portion of the organism's lipopolysaccharide. A wzy (O-antigen polymerase) deletion mutant of Ft. live attenuated vaccine strain (Ft.LVS), designated Ft.LVS::Δwzy, was created and evaluated as a live attenuated vaccine. Specifically, the mutant's virulence potential and its protective efficacy against type A and type B strains were investigated by challenge of immunized mice. F. tularensis LVS::Δwzy expressed only 1 repeating unit of O-PS and yet, upon immunization, induced O-PS-specific antibodies. Compared with Ft.LVS, the mutant was highly sensitive to complement-mediated lysis, significantly attenuated in virulence, and was recovered in much lower numbers from the organs of infected mice. Intranasal immunization with Ft.LVS::Δwzy provided protection against subsequent intranasal infection with the highly virulent type A strain SchuS4 and with Ft.LVS. Immunization with Ft.LVS::Δwzy elicited both humoral and cell-mediated immunity. Ft.LVS::Δwzy was avirulent in mice and, despite expressing only 1 repeating unit of the O-PS, induced antibodies to the full-length O-PS. Vaccination with Ft.LVS::Δwzy protected mice against intranasal challenge with both type A and type B strains of F. tularensis and induced functional immunity through both humoral and cellular mechanisms.