Fig. 2

CAR.CD123-NK cells exert a significant cytotoxicity against CD123⁺ AML cells. A–B CAR.CD123 is stably expressed on NK cells over extensive in vitro culture (evaluated up to 25 days). A The CAR.CD123 expression on NK cells from 9 different HDs was monitored at Day 3 and at Day 25 after transduction and is represented as average ± SD (B) Representative FACS analysis of CAR.CD123 expression in NK cells at Day 3 and Day 25 after transduction is shown. CAR expression was assessed by the use of anti-CD34 for the CAR detection in combination with anti-CD56 for NK detection. C Long-term 6-day cocultures were performed in 9 independent experiments, in which CD123⁺ leukaemia cell lines (namely, THP-1, OCI-AML3, and MOLM-13) (black bars) were cocultured with either NT-NK (grey bars) or CAR.CD123-NK cells (line black bars) derived from 9 independent HDs at the E:T ratio of 1:1. Data are shown as average ± SD. D Granz B, IFN-γ and TNF-α were measured by ELISA assay in 24 h culture supernatant of NT-NK (grey bars) or CAR.CD123-NK cells (line black bars) in response to CD123.⁺ leukaemia cell lines (namely, THP-1, OCI-AML3, and MOLM-13). Cytokine analysis was performed in 4 independent experiments, and data are shown as average ± SD. *p < 0.05, **p < 0.01, ***p < 0.001