Figure 5.

B6 mice (n=12) were vaccinated IN with 10,000 viable spores of Δcps1 or saline (n=8). Two weeks after vaccination splenocytes were harvested and stimulated with 10 ug/ml of Coccidioides spherule lysate (cocci stim) or PBS (NT) for 18 hours, then stained for T-cell cytokine production. There was a significant increase in CD4⁺ T cells producing IFN-γ in vaccinated mice stimulated with Coccidioides spherule lysate compared to unvaccinated mice (p=0.0008). A second set of vaccinated B6 mice (n=10) was infected with 100 spores of Cp 4 weeks post-vaccination. Two weeks post-infection, spleens were harvested and stimulated as above. As with vaccinated only mice, vaccinated and infected mice also had a significant increase in CD4⁺ cells making IFN-γ (p<0.0001). Calculation of the total number of CD4⁺ cells making IFN-γ (B) showed similar results for both vaccinated (p=0.0003) and vaccinated and infected mice (p<0.0001) compared to naïve animals. Dots for vaccinated only mice represent pools of 4 spleens; all other dots represent individual animals. Significance was determined using a Student’s t-test, and error bars indicate SD.