Figure 3.

Fine mapping of KMP11 and HASPB epitopes. A, Naive BALB/c mice were vaccinated in the footpad with 10⁹ pfu Ad5-KH and 10 days later splenocytes were restimulated with either the pooled pepsets from which the epitope was derived or synthetic peptides corresponding to predicted epitopes. After 3 h, Brefeldin A was added for an additional 4 h. Cells were surface-stained for CD3 and CD8 and then stained for intracellular IFNγ and IL-10. The percentage of cytokine positive cells is indicated in each quadrant. B, CD8⁺ T cells from mice vaccinated as above were restimulated with HASPB peptide, surface-stained for CD3 and CD8, and stained for intracellular cytokines. HASPB-specific CD8⁺ T cells were initially separated into IFNγ⁺TNF^-or IFNγ⁺TNF⁺ subsets and then further sub-divided on the basis of IL-2 and IL-10 production. Pie charts reflect the relative contribution of the different cytokine-producing CD8⁺ T cell populations and percentage of each population is indicated. Data are mean ± standard error of the mean (5 mice).