Figure 6.

Immunization with aggregated Ag85B results in expansion of polyfunctional T cells in immunized mice. T cells isolated from mice immunized with Bacillus Calmette–Guerin (BCG), Ag85B-IFA (F1), and aggregated Ag85B (F2) were stimulated ex vivo with corresponding antigens for 24 h and stained for intracellular IFN-γ and TNF-α. (A) Representative FACS dot plots denoting cumulative frequencies of CD4⁺ and CD8⁺ polyfunctional T cells. Cells co-expressing IFN-γ and TNF-α on immunization with BCG (a and a′), native Ag85B-IFA (F1) (b and b′), and aggregated Ag85B (F2) (c and c′). (B) Bar graphs depicting percent population of IFN-γ, TNF-α co-expressing CD4⁺ (a), and CD8⁺ (b) polyfunctional T cells. The cells were isolated from splenocytes of various immunized groups of mice at 1 week post booster. Dot plots are representative of at least three animals for each group. The data were analyzed by employing one-way ANOVA followed by Bonferroni’s multiple comparison test and shown as means (±SD) of three independent determinants, where p < 0.05(*), p < 0.01(**) and p < 0.001(***) were considered significant.