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. 2017 Feb 6;127(3):929–941. doi: 10.1172/JCI89455

Figure 6. Targeting the A2AR with shRNA retroviral technology enhances CAR T cell function.

Figure 6

CAR T cells were transduced with retroviruses encoding A2AR-directed shRNA or scrambled shRNA control and then selected with 2 μg/ml puromycin from day 4 to day 6. Eight days after activation, CAR T cells were then assessed for function. (A) 1 × 106 CAR T cells were reactivated with plate-bound anti-CD3/anti-CD28 (0.5 μg/ml) for 4 hours and then lysed for RNA analysis. Data are presented as the mean ± SD of triplicates and expressed relative to the production of IFN-γ of control cultures for each shRNA-transduced CAR T cell. (B) 1 × 105 CAR T cells were cocultured with 24JK/24JK-HER2 tumor cells at a 1:1 ratio in the presence or absence of NECA at indicated concentrations. Supernatants were collected after 16 hours and analyzed for concentration of IFN-γ. Data are presented relative to the IFN-γ production of control cultures and as the mean ± SD of quadruplicates from a representative experiment of n = 2. *P < 0.05, **P < 0.01 by 1-way ANOVA.