Fig. 2

Efficient A2aR knockdown in MSLN-CAR T cells by anti-A2aR shRNA. a Human peripheral T cells, Mock T, as well as 4 different groups of CAR T cells from 6 healthy donors were analyzed to determine the percentage of A2aR positive cells. All cells were evaluated before and after stimulation with indicated stimulators. Histogram plots represent percentage of A2aR positive T cell, Mock T cell and CAR T cells from one donor. b Bar graph represents the average value for A2aR positive peripheral T cells, Mock T cells and CAR T cells from all 6 donors with or without stimulation. c Quantification of mean fluorescence intensity (MFI) demonstrates a reduction in A2aR expression in A2aR.KD1, KD2, and KD3.MSLN-CAR T cells from 6 donors (MFI for MSLN-CAR T cell was set as 100%). * Intragroup comparison of A2aR positive cells before and after stimulation. # Intergroup Comparison of the average levels of the percentage of A2aR positive cells in MSLN-CAR T cell groups with A2aR.KD1, KD2 and KD3.MSLN-CAR T cell groups. ɸ Intergroup Comparison of the percentage of A2aR positive cells in stimulated T cells with stimulated MSLN-CAR T cell groups. P < 0.05 were considered as statistically significant calculated with one-way ANOVA with Tukeyʼs post-test for multiple comparisons. MSLN-CAR: fully human anti mesothelin CAR, A2aR-KD-MSLN-CARs: A2aR-knocked down anti mesothelin-CAR VCM: viruses containing media; MFI: mean fluorescence intensity. (*P < 0.05, ^ɸP < 0.05, ^{ɸ ɸ}P < 0.01, ^***P < 0.001, ^###P < 0.001)