Figure 4.

CE-XTC manufacturing induces CE Env- and CE Gag-specific CD4+ and CD8+ T cells expressing one or more effector functions. Pre-expansion PBMC (circles) and post-expansion CE-XTCs (triangles) from SHIV-infected, ART suppressed donors (closed shapes, n = 3) and uninfected donors (open shapes, n = 2) were expanded for 19 days and then stimulated overnight with CE peptide pools comprising either 7 regions of SIV Gag (A, C) or 12 regions of HIV Env (B, D). Following peptide stimulation, effector functions were analyzed by intracellular cytokine staining and flow cytometry. Shown are the increases in frequencies of CD4 and CD8 T cells expressing one or more of the cytokines IFN-γ, IL-2, TNFα or co-expression of CD107a and Granzyme B as markers of cytolytic effector function after the 19-day expansion period. P values comparing responses before and after expansion are shown for each individual effector function (A, B) or as polyfunctional responses (C, D) defined as cells expressing 1 or more cytokine and/or cytolytic effector functions. Individual responses were compared by Mixed-effects model (REML) adjusted for multiple comparisons by Sidak’s multiple comparison test (A, B) * = p<0.05. Differences in paired pre- vs. post-expansion T-cell response polyfunctionality were compared by Wilcoxon matched-pairs signed rank test (C, D).