Fig. 4.

Targeting chronic lymphocytic leukemia (CLL) with MC10029 CAR-T cells. a Flow cytometry contour plots show the functional potency of CAR-T cells against target cells by the surface expression CD107a in a degranulation assay. Non-CAR-T cells and MC10029 CAR-T cells were generated from the same donor and incubated with MEC-1 cells at an E:T ratio of 2:1 to characterize the cytotoxicity of MC10029 CAR-T cells. (Statistical analysis, Supplementary Fig. 7e). b A granzyme B ELISA shows functional potency of MC10029 CAR-T cells against MEC-1 cells. Non-CAR-T cells or MC10029 CAR-T cells were co-incubated MEC-1 cells at an E:T ratio of 4:1 for 72 h; the supernatants were then harvested for subsequent ELISA. Graphed data are means of quadruplicate sampling. The data are representative of three independent experiments. c Flow cytometry shows the cell surface expression of BAFF-R on enriched tumor cells and PBMCs that were collected from six subjects with CLL. d MC10029 CAR-T cells derived from two different healthy donors were incubated with the enriched primary B-cell tumor cells at an E:T ratio of 2:1. Using a CD107a degranulation assay, the cytotoxic activity of MC10029 CAR-T cells was visualized using flow cytometry. Non-CAR T cells from the same donor were used as a negative control. e The release of granzyme B confirms the functional potency of MC10029 CAR-T cells against primary CLL tumor cells. MC10029 CAR-T cells or non-CAR-T cells were co-incubated with enriched primary CLL tumor cells (E:T ratio of 4:1) for 72 h, after which the supernatants were harvested for subsequent ELISA. Graphed data are means of quadruplicate sampling. (***p < 0.001)