Figure 4.

Phenotype and function of human T cells expanded with PIC immunobrush beads on day 14. (a) Ratio of CD4⁺–CD8⁺ human T cells after expansion n = 2–5 in one or two independent experiments. Data were analyzed using a mixed-effects model, followed by a Dunnett’s multiple-comparisons test on log2-transformed data. (b, c) CD4⁺ and CD8⁺ differentiation of human T cells in to different memory subsets after expansion with PIC beads, Dynabeads, or T Cell TransAct, respectively; n = 2 in one independent experiment. (d) CD4⁺ and (e) CD8⁺ cytotoxic character of human T cells after expansion; n = 5 in two independent experiments. Statistics: CD4⁺ T cells: CD107^–GZMB⁺ PIC beads vs Dynabeads p < 0.05. CD8⁺ T cells: CD107^–GZMB^– PIC beads vs Dynabeads p < 0.05, CD107^–GZMB⁺ PIC beads vs T cell Transact p < 0.01, CD107⁺GZMB⁺ PIC beads vs T cell Transact p < 0.05. (f, g) CD4⁺ and CD8⁺ exhaustion character of human T cells after expansion, respectively; n = 5 in two independent experiments. Statistics: CD4⁺ T cells: PD-1^–TIM3^– PIC beads vs T cell Transact p < 0.01, PD-1^–TIM3⁺ PIC beads vs T cell Transact p < 0.05, PD-1⁺TIM-3⁺ PIC vs T cell Transact p < 0.01. CD8⁺ T cells: PD-1^–TIM3⁺ PIC beads vs T cell Transact p < 0.01, PD-1⁺TIM3⁺ PIC vs T cell Transact p < 0.01. (h, i) Intracellular expression of 0, 1, 2, or 3 cytokines (IL-2, IFNγ and/or TNFα) in CD4⁺ (h) and CD8⁺ (i) T cell; n = 5 in two independent experiments. Statistics: CD4⁺ T cells: three markers PIC vs Dynabeads p < 0.05. CD8⁺ T cells: 1 marker PIC vs T cell Transact p < 0.05, 0 markers PIC vs T cell Transact p < 0.05. (b–i) Significance was analyzed with an RM two-way ANOVA with Geisser–Greenhouse correction, followed by Dunnett’s multiple-comparisons test.