Supplemental Figure 2 from Augmenting CAR T-cell Functions with LIGHT

(A) ELISA quantification of soluble LIGHT in cell culture media after 24 hours of incubation. (B) Supernatant from LIGHT-CAR T cells and second-generation CAR T cells were added to corresponding CAR T cells and in vitro cytolysis was assessed against AsPC1. Data is representative of 2 independent experiments from two different T cell donors. (C) Supernatant from LIGHT-CAR T cells and second-generation CAR T cells were added to corresponding CAR T cells and in vitro cytolysis was assessed against MIAPACA2. Data is representative of 2 independent experiments from two different T cell donors. (D) Cytotoxicity assay with various mesothelin-directed CAR constructs and the addition of recombinant LIGHT to MIAPACA2. Data is representative of 3 independent experiments from two different T cell donors. (E) Mesothelin-directed LIGHT-CAR T cells exhibited similar proliferation in a repetitive antigen stimulation assay with moderately high mesothelin expression PDAC cell line, AsPC1. CAR T cells were cocultured with tumor cells at a 4:1 effector: tumor ratio for 5 days, and then CAR T cells were taken out and put onto new tumor cells at the original E:T ratio. The total fold expansion was quantified from multiplying each round of fold expansion every 5 days. Data is representative of 3 independent experiments of 3 different human donors and data errors were analyzed with mean ± SEM.