Figure 4.

Review of activation marker CD69 and CD25 expression by CD4⁺ and CD8+ T cell populations after CD3/CD28 bead-based activation in a 100-mL culture bag

(A) Early activation (CD69⁺CD25⁻), (B) late activation (CD69⁻CD25⁺), and (C) peak activation (CD69⁺CD25⁺) by CD4⁺ and CD8⁺ T cells after 48-h Dynabead activation of unsorted leukapheresis, monocyte-depleted, and isolated T cell starting populations. 5 different donors were activated: donor 1 (◇), donor 2 (○), donor 3 (⨯), donor 4 (▵), and donor 5 (▹). Donors 2, 4, and 5 were replicated for subsequent CAR viral transduction alongside donor 9 (denoted donor 2_CAR [●], donor 4_CAR ▴], donor 5_CAR [▸], and donor 9_CAR [★]). Error bars represent the mean ± 1 SD, where the mean of the value is represented by □. Statistical significance was determined by one-way ANOVA and Dunnett’s hypothesis testing; ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001.