BALB/c mice were immunized with rBCG-Mkan85B followed by boosting with DNA-Mkan85B three times. Spleen cells from the immunized mice were stimulated with pep8 and pep9 peptide. (A) Different dose-response-generation of polyfunctional CD8+ T cells by the epitope peptides. pep8 is much more functional for the CD8+ T cell induction than pep9 stimulation of immune T cells by comparing the 50% stimulation dose between the functional pep8 and pep9. Data represent three independent experiments with three mice per group. Error bars represents SEM. (B) Representative histograms of CD107a/b expression. The polyfunctional and effector cytokines-producing CD8+ T cells were analyzed for expression of CD107a/b. Percentages of maximal expression with CD107a/b (% of max) are presented in histograms of red line (polyfunctional CD8+ T cells of pep8 or pep9-stimulations, left and right panels, respectively) and of blue line (polyfunctional) CD8+ T cells of simultaneous stimulation with PMA and ionomycin (P+I)), which were mainly negative of CD107a/b expression. (C) Expression of CD107a/b on the polyfunctional CD8+ T cells. Data are presented as the percentage of the CD107a/b expression in the polyfunctional cells (closed bar), and CD107a/b negatively (open bar) in the P+I, pep8 or pep9 -stimulated polyfunctional CD8+ T cells. Data represent three independent experiments with five mice per group. Error bars represent SEM.