Figure 3: A) Flow cytometric analysis of pentamer binding (HLAA* 0201) to human survivin-specific CD8+ T cells stimulated ex vivo with SurVaxM. Specific reactivity of CD8+ T cells to two different survivin epitopes but not control peptide are shown (upper right quadrants are CD8+/Pentamer+). B) Flow cytometric measurement of CD4+ proliferation via CFSE dye mitotic distribution in response to ex vivo stimulation with SurVaxM. As CD4+ cells proliferate, CFSE intensity decreases as dye is redistributed among daughter cells. Stimulation was without added peptide (Control) or with SurVaxM peptide. Grey, raw data; black line, computer curve fit; blue, undivided cells; yellow line, multiple proliferative generations. Data acquisition using BD LSRFortessa and analysis using FCS express v4. [anti-survivin custom pentamer (ProImmune, Oxford, UK) and CFSE (Invitrogen-Molecular Probes, Eugene, OR)].