Fig. 3: A_2AR knockdown promotes an effector-like CAR T-cell phenotype in vivo.

C57BL/6 Her2 mice were injected with 2 × 10⁵ E0771-Her2 tumor cells into the fourth mammary fat pad and treated with anti-Her2 CAR T cells transduced and cultured in IL-7 and IL-15 as per Fig. 2G. Once tumors were established, 1 × 10⁷ CAR T cells were injected intravenously on two subsequent days following 4-Gy total body irradiation. Mice were treated with 50,000U of IL-2 on days 0–4 post CAR T-cell treatment. A Tumor growth shown as the mean ± SEM of n = 5 (CAR T-cell treated) or 6 (nontreated) mice per group from a representative experiment: n = 2, *p < 0.05, two-way ANOVA. B–E Spleens and tumors were excised 9 days post treatment and the number and phenotype of NGFR⁺ CAR T cells determined by flow cytometry. B, D Expression from concatenated samples (n = 7 per group) from a representative experiment. C, E Data represent the mean ± SEM of n = 11–19 mice⁺ from 2 (E) or 3 (C) replicate experiments (see ⁺ below for details). Each data point represents an individual mouse. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, unpaired t test. ⁺ IFNγ, TNF, TIM-3 control n = 15, shRNA A_2AR n = 18, Granzyme B and Ki-67, control n = 15, shRNA A_2AR n = 19, IRF4, control n = 16, shRNA A_2AR n = 18, numbers in spleen control n = 11, shRNA A_2AR n = 13. Data from some control samples are also presented in ref. ³⁶. Source data are provided as a Source Data file.