Dose of Incorporated Immunodominant Antigen in Recombinant BCG Impacts Modestly on Th1 Immune Response and Protective Efficiency against Mycobacterium tuberculosis in Mice

Figure 3

Flow cytometric analysis of intracellular cytokine production after immunization. Eight weeks after vaccination, splenocytes from 6 mice were isolated, pooled and stimulated with PPD for 14 h, and then analyzed for cytokine production by intracellular cytokine staining (ICS) assay. CD3⁺CD4⁺ T cells producing IFN-γ, TNF-α, and IL-2 were distinguished as seven distinct subpopulations based on their production of these cytokines in any combination. The subpopulation proportions as components of the total CD4⁺ T cell population are shown (a) and their proportions as components of the seven subpopulations are shown in pie chart form (b). Integrated mean fluorescence intensities (iMFI) of IL-2 in four cytokine profiles (c). Data are shown as mean ± SEM. ^∗∗𝑃<0.01 (one-way ANOVA).