Figure 3.

GD2-CAR T cell killing assay. A. CAR T and target cell coculture and analysis of cytolytic activities by flow cytometry. B. Target cell killing analysis of the three different GD2 CAR T cells, Hu3F8, Hu14.18 and c60C3. The OS cells transduced with a lentiviral wasabi green fluorescence reporter gene were used as target cells, and plated into 96 or 48 wells for 2 hours before adding the different CAR T cells (Jurkat or primary T cells, at E:T ratio 3:1). After co-culture at 37°C, the cells were harvested using 2.5-5 mM EDTA, washed twice with PBS and stained with Annexin V and propidium iodide (PI). The percentage of target cell death was detected by flow cytometry and the % specific lysis was calculated as illustrated.