CAR-ML NK cells exhibit enhanced functional responses compared with CAR cNK cells. cNK cells or ML NK cells were transduced with 19-CAR lentivirus and effector functions against CD19⁺ Raji targets were assessed 7 days later. (A) Representative GFP gating strategy from cNK (blue) and ML NK cells (green) transduced with 19-CAR lentivirus. (B) Representative flow plots of GFP⁺ cNK or ML NK cells stimulated with Raji targets (total NK/T, 5:1) depicting IFN-γ (top) and degranulation (CD107a; bottom). (C) Summary IFN-γ production from panel B. (D) Summary degranulation from panel B (n = 8-9; 5 independent experiments). (E) Combination index values of IFN-γ and CD107a from panels C and D. (F) Summary data show mean ± SEM percentage of specific killing by 19-CAR cNK and 19-CAR-ML NK cells at the indicated total NK effector/target ratios (n = 4; 2 independent experiments). cNK cells, open lavender circle; ML NK cells, orange circle; 19-CAR cNK cells, blue open triangle, 19-CAR ML NK cells, green triangle. Data were compared using 2-way analysis of variance with Bonferroni’s multiple comparisons test (C-E) or linear mixed model with Bonferroni’s multiple comparisons test (F). *P < .05, **P < .01, ***P < .001, ****P < .0001. ns, no significance.