FIGURE 5.

CARD9 deficient mice have a reduced protective efficacy concurring with reduced Th17 response to the GCP-rCpa1 vaccine. WT and Card9^−/− (ΔC9) mice (n=10) were subcutaneously immunized with GCP adjuvant alone or GCP-rCpa1 vaccine twice and then intranasally challenged with a lethal dose of Coccidioides spores. (A) Fungal CFUs were detected by dilution plate culture of lung and spleen homogenates at 14 dpc. CFUs are reported in box plots and asterisks indicate statistically significant differences (Mann-Whitney U test; p <0.05) between the vaccinated and nonvaccinated mice of the same strain, while the hashtags indicate significant differences between these 2 strains of mice (Mann-Whitney U test, P< 0.05). (B) Representation plots of gated IL-17a- and IFN-γ-expressing Th17 and Th1 cells derived from the lungs of WT and ΔC9 mice were analyzed by the intracellular cytokine staining method. (C) Data were processed using FlowJo X software. Total numbers of Th1 and Th17 cells were determined for both vaccinated WT and ΔC9 mice at 7 and 14 dpc and compared to those of nonvaccinated mice. Data are mean values ± SEM of 3-4 mice per group. For statistical analysis of cell population, asterisks indicate significantly higher percentage (B) and cell numbers (C) of the responsive T-cell phenotypes in lungs of vaccinated compared to nonvaccinated mice of the same strain, while hashtags represent significant differences in the vaccinated ΔC9 mice compared to WT mice (Mann-Whitney U test, P< 0.05).