Enhanced Short- and Long-Term Reactivity of CAR19-iNKT Cells against B Lineage Malignancies (A) Second- and third-generation CAR19-T and CAR19-iNKT cell expansion (fold change) and absolute cell numbers (cell count) over a period of 3 weeks (n = 4). p value is for CAR19-iNKT versus CAR19-T cells using Friedman test. Error bars represent SEM. (B) Proliferation analysis of second- and third-generation CAR19-T and CAR19-iNKT cells in the presence (stimulated) or not (resting) of irradiated CD1d + CD19 + (C1R-CD1d) cells over 7 days. p value is for CAR19-iNKT versus CAR19-T cells using Friedman test. (C) Cytotoxicity of third-generation CAR19-T and -NKT cells against C1R-CD1d (representative of n = 3) and Farage lymphoma cell lines (representative of n = 2) pre-loaded or not with αGalCer. Error bars represent SEM of triplicate assays. (D) IncuCyte images of representative wells showing the final effector (gray) and live target cells (red) after 7 days. Effectors were second-generation CAR19-T and CAR19-NKT cells. Targets were CD19 + ARH-77-CD1d cells expressing mCherry red fluorescent protein. Scale bar represents 400 μm. (E) Seven-day trajectory of effector and target cell proliferation and elimination respectively as per (D). p value is for CAR19-iNKT versus CAR19-T cells using Friedman test. (F) Cytotoxicity of second-generation CAR19-iNKT, CAR19-T, and of untransduced iNKT cells against lymphoma cells from one patient with mantle cell lymphoma (MCL; top) and two patients with marginal zone B lymphoma (MZL; bottom) using three different T/iNKT cell healthy donors (A, B, and C). Error bars represent SEM of triplicate assays. ∗∗ p