To evaluate for autonomous NK cell growth, we maintained control NT and iC9/CAR.19/IL15+ NK cells in SCGM without stimulation or addition of exogenous cytokines. Cells were cultured for 42 days, and counted using trypan blue exclusion every 3 days.
To assess cytotoxicity, CAR-transduced and NT-NK cells were cocultured with 51Cr-labeled CLL, Raji and K562 targets (positive control) at multiple E:T ratios; cytotoxicity was measured by 51Cr-release as previously described,(14 (link)) the findings are reported as specific lysis relative to K562 targets.(24 (link)) For HLA-blocking experiments, the anti-HLA-ABC clone W6/32 (Biolegend) was used.
To assess cytotoxicity, CAR-transduced and NT-NK cells were cocultured with 51Cr-labeled CLL, Raji and K562 targets (positive control) at multiple E:T ratios; cytotoxicity was measured by 51Cr-release as previously described,(14 (link)) the findings are reported as specific lysis relative to K562 targets.(24 (link)) For HLA-blocking experiments, the anti-HLA-ABC clone W6/32 (Biolegend) was used.
