Dual-specific cytotoxicity of CAR.GD2 NKT cells. The cell-mediated cytotoxicity of parental and CAR.GD2 NKT cells with the indicated constructs was tested using a 4-hour ⁵¹Cr-release assay against the following targets: GD2⁺CD1d^neg CHLA255 NB cell line (A); GD2^negCD1d⁺ in vitro polarized M2 macrophages (B); and GD2^negCD1d^neg LA-N-6 NB cell line (C). (D) 7-AAD staining was performed on day 3 of NKT/NB cell culture. Histograms show percent 7-AAD positive cells after gating on CAR-positive cells with the indicated constructs. Results are from a representative of 3 independent experiments.