Interaction of ND-XNK cells with CLL targeted therapies. (A) ND-XNK cells were cocultured with primary CLL cells and 5 μg/mL obinutuzumab ± 1 μM ibrutinib, 1 μM acalabrutinib, 5 μM idelalisib, or 8 nM venetoclax. Cytotoxicity measured by 4-hour calcein release assay. Graph shows mean ± standard deviation of 10 to 12 experiments. (B) Left: ND-XNK cells were stimulated for 3 days with 10 μg/mL IL-2 and/or IL-15, plus IL-12 to induce AICD, ±1 μM ibrutinib or acalabrutinib. Survival was measured as the % of annexin V^–/PI^– cells. Graph shows mean ± standard deviation of 5 experiments, normalized to each stimulus without IL-12. Right: ND-XNK cells were stimulated for 3 days with plate-adsorbed rituximab or obinutuzumab, ±1 μM ibrutinib or acalabrutinib. Survival was measured by annexin V/PI staining and shown as mean ± standard deviation of 5 experiments, normalized to the unstimulated condition to account for normal cell death in culture. (C) Expression of BTK, RLK, TEC, and ITK was measured in NNKs and ND-XNKs by western blotting. (D) ND-XNK or NNK cells were cultured with varying concentrations of venetoclax for 24 hours, and viability was measured by annexin V/PI flow cytometry. Graph shows mean ± standard deviation of 4 NNK, 5 ND-XNK experiments. (E) Expression of Bcl-2, Mcl-1, Bcl-XL, and Bim was measured in NNKs and ND-XNKs by western blotting. ∗P < .05 by mixed effect modeling. mAb, monoclonal antibody.