Figure 6. IFNγ secretion and granzyme B up-regulation triggered by tumor antigen-specific recognition of anti-EpCAM RNA CARs.
(A and B) Increased IFNγ secretion as determined by IFNγ ELISPOT assay. T cells were electroporated with anti-EpCAM RNA CARs and co-cultured with target tumor cells overnight before assay. mGFP RNA CAR-transfected T cells served as a negative control. Results from EpCAM-negative tumor cell lines Raji, IGR1, and PA-1 and EpCAM-positive cancer cell lines SKOV3-Luc and CAOV-3 are shown in (A) and results from EpCAM-positive cancer cell lines HCT8, HRT-18G, SW480, SW620 and SW626 in (B). (C) Granzyme B up-regulation as determined by granzyme B ELISPOT assay. EpCAM-positive tumor cell lines HRT-18G and SKOV3-Luc were tested as described in (A and B) Mean IFNγ or granzyme B spots per 1 × 106 T cells ± SD from triplicate cultures are shown in A, B and C.