Fig. 4.

In vitro functionality of CAR T and CAR NK cells. Expanded cells were cocultured with different cancer cell lines at an effector to target (E:T) ratio of 5:1 for 4 h. Specific killing was assessed by flow cytometry using a membrane-impermeable DNA stain (TO-PRO-3) and PKH26 membrane-labeled target cells and IFN-γ production was measured by ELISA. A Schematic of the timeline. B, C Specific (left) and CAR-mediated (right) killing of CD19⁺ 721.221 (B) and CD19^KO 721.221 cells (C) by anti-CD19 CAR T, CAR NK cells or untransduced controls. D Overall (left) and CAR-mediated (right) IFN-γ production by anti-CD19 CAR T, CAR NK cells and untransduced controls after coculture with CD19⁺ 721.221 cells. (E, F) Specific (left) and CAR-mediated (right) killing of NALM6 (E) or Raji (F) cells by anti-CD19 CAR T, CAR NK cells and untransduced controls. G, H Overall (left) and CAR-mediated (right) IFN-γ production by anti-CD19 CAR T, CAR NK cells and untransduced controls after coculture with NALM6 (G) or Raji (H) cells. I Specific (left) and CAR-mediated (right) killing of LCL by autologous anti-CD19 CAR T cells, allogeneic anti-CD19 CAR NK cells and untransduced controls. J Overall (left) and CAR-mediated (right) IFN-γ production by autologous anti-CD19 CAR T cells, allogeneic anti-CD19 CAR NK cells and untransduced controls after coculture with LCL target cells. Data are from 1 to 3 independent experiments with n = 3–12 donors. Significance by mixed-effects analysis, RM one-way ANOVA with Tukey’s multiple comparisons, ordinary one-way ANOVA with Tukey’s multiple comparisons, two-tailed paired t-test or two-tailed unpaired t-test as appropriate; *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. Symbols represent individual donors. Data shown as box and whiskers display the median as a line within the box; whiskers are shown from minimum to maximum data point