File S1 - A ΔclpB Mutant of Francisella tularensis Subspecies holarctica Strain, FSC200, Is a More Effective Live Vaccine than F. tularensis LVS in a Mouse Respiratory Challenge Model of Tularemia

Figure S1. Attenuation of FSC200ΔclpB. BALB/c mice (n = 5/group) were challenged ID with 107 CFU of FSC200ΔclpB (circle) or 10 CFU of complemented FSC200ΔclpB (squares), and were monitored for survival. Figure S2. Intranasal virulence of FSC200ΔclpB versus LVS. BALB/c mice were inoculated IN with 104 (open circle) or 105 (closed circle) CFU of FSC200ΔclpB or 103 (open square) or 104 (closed square) CFU of LVS and were monitored for survival. *, significantly shorter survival than mice challenged with the same dose of FSC200ΔclpB. Figure S3. In vivo growth kinetics of FSC200ΔclpB vs LVS. BALB/c mice (n = 4/group) were immunized ID with ∼ 105 CFU of FSC200ΔclpB (circle) or LVS (square). Mice were killed on days 2,4,7, and 14 for bacteriology. *, significantly higher burden vs LVS. Dashed line shows limit of detection. Figure S4. Skin reactogenicity score chart for F. tularensis strains. Figure S5. Protection against ID or IN challenge with SCHU S4 following immunization with SCHU S4ΔclpB versus LVSΔclpB. BALB/c mice were immunized ID with ∼105 CFU of SCHUS4ΔclpB or LVSΔclpB. Immunized and control mice were challenged six weeks later with 1000 CFU ID or 40 CFU IN of SCHU S4 and were monitored for survival. *, significantly longer survival than control mice; **, significantly longer survival than mice immunized with LVSΔclpB. Table S1. Growth of SCHU S4ΔclpB, FSC200ΔclpB, and LVS in the tissues of SCID mice. Table S2. Attenuation and immunogenicity profiles of selected SCHU S4 deletion mutants administered ID or IN to BALB/c mice. (PDF)