Figure 3
Effector CD8+ T cells generated under TSCM-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 TSCM-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 TSCM-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 TSCM-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).

Effector CD8+ T cells generated under TSCM-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 TSCM-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 TSCM-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 TSCM-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).

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