Effector CD8⁺ T cells generated under T_SCM-enriched culture condition are polyfunctional. (A) Concentration of cytokine in the supernatant of CD19-CAR–modified T cells after 16-hour coculture with CD19⁺ SUDHL4 cells or CD19⁻ CCRF-CEM cells. Values are shown after subtraction of background cytokine release values obtained after coculture with the CCRF-CEM. Data representing results from 6 healthy volunteer donors (HDs) are shown as box-and-whisker plots extending to minimum and maximum values. Bands inside the box represent median values (*P < .05; Wilcoxon matched-pairs signed rank test). (B) Intracellular cytokine staining of CD19-CAR–modified standard and T_SCM-enriched products from a representative HD after coculture with CD19⁺ SUDHL4 cells. Data are shown after gating on live CD3⁺CD8⁺ cells. Numbers indicate the percentage of cells in each quadrant. (C) Mean fluorescence intensity of cytokines produced by CCR7⁻ effector CD8⁺ T cells within standard and T_SCM-enriched products after coculture with CD19⁺ SUDHL4 cells. Data representing results from 6 HDs are shown as box-and-whisker plots extending to minimum and maximum values. Bands inside the box represent median values (*P < .05; Wilcoxon matched-pairs signed rank test). (D) Intracellular cytokine staining of CD19-CAR–modified standard and T_SCM-enriched products from a representative HD after coculture with CD19⁺ SUDHL4 cells. Data are shown after gating on live CD3⁺CD8⁺CCR7⁻ cells. Numbers indicate the percentage of cells in each quadrant. (E) Pie charts depicting the quality of the cytokine response in CCR7⁻ effector CD8⁺ T cells from 6 HDs after coculture with CD19⁺ SUDHL4 cells. Values are determined by the Boolean combination of gates identifying IFN-γ⁺, IL-2⁺, TNF-α⁺, and CD107a⁺ cells. Numbers indicate cell percentages. (F) Percentage of polyfunctional CCR7⁻ effector CD8⁺ T cells from 6 HDs after coculture with CD19⁺ SUDHL4 cells (*P < .05; Wilcoxon matched-pairs signed rank test).