Fig. 3

GUCY2C-specific CD8⁺, but not CD4⁺, T-cell responses. PBMCs from GUCY2C-responder patients 1007 (a and b) and 1008 (c and d) collected 30 days after Ad5-GUCY2C-PADRE administration were left unsorted or depleted of CD4⁺ or CD8⁺ T cells by MACS. PBMCs, CD4-depleted PBMCs, and CD8-depleted PBMCs were analyzed by FACS to confirm depletion (a and c) and tested for GUCY2C-specific T-cell responses by IFNγ-ELISpot (b and d). NS = not significant, *** P < 0.001, **** P < 0.0001, Two-way ANOVA. Depletion efficiencies determined by FACS were > 98% for CD4⁺ T cells and ~ 75–95% for CD8⁺ T cells