Mice were immunized with saline or liquid, liquid covial, or lyophilized covial ID93+GLA-SE exposed to 4°C or 50°C for one month. (A) B and T cell blood counts were determined 18 hours after immunization. (B) ID93-specific serum antibody titers were determined three weeks after the first immunization. (D) The frequency of ID93-specific CD4 T cells in the spleen were evaluated one month after the final immunization by analyzing cytokine production following in vitro restimulation with ID93. One month after the final immunization animals were challenged with a low dose of aerosolized M. tuberuculosis. Bacterial burden in the (D) lung and (E) spleen were determined three weeks later. Data are displayed as mean + s.d. of N=5 −7 mice/group. Data are shown from one of two experiments with similar results. *, **, ***, and **** indicate P<0.05, 0.1, 0.001 and 0.0001, respectively, relative to saline. n.s. not significant relative to saline. Statistical comparisons between 4ºC and 50ºC samples are indicated.