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. 2021 Jan 18;12:409. doi: 10.1038/s41467-020-20696-x

Fig. 3. dCAR T cells exhibit enhanced antitumour reactivity and the maintenance of a memory-like phenotype at a low effector:target ratio.

Fig. 3

a Continuous graphical output of cell index values determined using the xCELLigence Impedance system from NT, CAR T and dCAR T cells co-cultured with Raji cells at an effector-to-target (E:T) ratio of 1:5 up to the 140-h time point. The density of Raji cell changes from baseline at each time point was calculated and compared using two-tailed paired t tests. **P < 0.01. (n = 2). b Representative phase: dCAR T or CAR T cells co-cultured with Raji cells at an E:T ratio of 1:30 (n = 2). Green fluorescent (nuclear-restricted RFP) cells were CAR T or dCAR T cells. Red fluorescent (Yoyo3) cells were counted as dead cells. Scale bar: 400 μm. c Cytotoxic analysis of dCAR and CAR T cells co-cultured with Raji cells at an E:T ratio of 1:1 to 1:30 (1:1, 1:5, 1: 10 (n = 4); E:T = 1:20 and 1:30 (n = 3)). d CD107a expression on CAR T cells after co-culture with Raji cells at an E:T ratio of 1:1 for 0.5 or 1 h (n = 3). e Cytokine production by dCAR and CAR T cells co-cultured with Raji cells at an E:T ratio of 1:1 for 24 h (n = 4). f Histogram plots of HLA-DR and Ki67 expression on of CAR and dCAR T cells after co-culture with Raji cells at an E:T ratio of 1:1 for 24 h (n = 3). g Pattern of repeated antigen stimulation in vitro. (“↓” represents the time point when Raji cells were added). h Cell count analysis of CAR T-cell proliferation as measured after 24 h of co-culture with Raji cells at every time point (n = 5). i Continuous testing of CD3/CD25, CD3/EOMES, CD3/PD1 and CD45RO/CD62L expression on CAR and dCAR T cells was performed after 24 h of co-culture with Raji cells at an E:T ratio of 1:1 (n = 3). Data for all panels are presented as the mean ± s.e.m. P values for c–f, h and i were calculated by two-tailed unpaired t tests. *P < 0.05, **P < 0.01, ***P < 0.001 and ****P < 0.0001. For c and e, exact P values are available in the Source Data File. Source data are provided as a Source Data file.