Figure 2

Specific IFN-γ and IL-2 release and proliferation of PSCA-CAR-engineered T cells. Lymphocytes were isolated from healthy donors and T cells were stimulated before being transduced with a lentiviral vector encoding the CAR against PSCA or Mock lentivirus. PSCA-CAR T cells were then co-cultured overnight with mel526(PSCA) or irrelevant mel526(TARP) target cells. In a separate experiment PSCA-CAR T cells and Mock T cells were co-cultured with relevant target mel526(PSCA) cells. ELISA was used to analyze IFN-γ release from (A) PSCA-CAR T cells against mel526(PSCA) or mel526(TARP) target cells (n = 4), (B) PSCA-CAR T cells and Mock T cells against mel526(PSCA) target cells (n = 4). ELISA was used to analyze IL-2 release from (C) PSCA-CAR T cells against mel526(PSCA) or mel526(TARP) target cells (n = 4), (D) PSCA-CAR T cells and Mock T cells against mel526(PSCA) target cells (n = 4). PSCA-CAR T cells and Mock T cells were labeled with a fluorescent dye and T cell proliferation, assessed as dilution of the dye for each cell division, was measured by flow cytometry after 5 days of co-culture with mel526(PSCA) target cells or mel526(TARP) control cells. Pooled T cell proliferation data for (E) PSCA-CAR T cells against mel526(PSCA) or mel526(TARP) target cells, (F) PSCA-CAR T cells and Mock T cells against mel526(PSCA) target cells (n = 4). (G) A representative histogram of proliferation for T cells established from one donor in the pool presented in E. (H) A representative histogram of proliferation for T cells established from one donor in the pool presented in F. Asterisks indicate significance (***p < 0.001, paired Student’s t-test). Error bars represent standard deviation from four individual donors run in triplicates.