Fig. 3: Antigen sensitivity and KRAS-G12V specific tumor responsiveness of 1-2C and 3-2E TCR-T cells.

a Schematic of the functional evaluation assay of the TCRs in Jurkat T cells, created with BioRender.com. Jurkat T cells were transduced to express 1-2C (b) or 3-2E (c) engineered Jurkat T cells were subsequently co-cultured with K562-HLA-A11 target cells and serially diluted peptides for 24 h. Co-cultured supernatants were analyzed by ELISA for secreted IL-2. d Schematic of the functional evaluation assay of the TCR-T cells engineered with primary T cells, created with BioRender.com. e, f 1-2C or 3-2E TCR-T cells were co-cultured with PANC-1 target cells and serially diluted peptides for 24 h. Co-cultured supernatants were analyzed by ELISA for secreted IFN-γ. Each data point represents the mean concentration of IL-2/IFN-γ for each sample run in triplicate wells in (b), (c), (e) and (f), and the data are representative of n = 3 independent experiments. g The 1-2C TCR-T cells were co-cultured with PANC-1 cells stably expressing the wildtype KRAS genes or KRAS G12V, G12C, or G12D mutants. Responses of 1-2C TCR-T cells with wildtype CFPAC-1 cells or CFPAC-1 cells stably expressing HLA-A*11:01 (h), or with wildtype SW-620 cells or SW-620 cells stably expressing HLA-A*11:01 (i). j–l Reactivity of 3-2E TCR-T cells against PANC-1, CFPAC-1, and SW-620 as indicated in (g–i). Each dot of (g–l) represents one technical replicate (n = 1 experiment). The responses were evaluated with 1-2C or 3-2E TCR-T cells prepared with T cells from n = 3 separate donors run independently (Donor 1-3) from (g) to (l). The columns show means of the three technical replicates. (m-r) Luciferase-transduced cell lines were co-cultured with mock-T or TCR-T for 48 h at various E: T ratios. The % specific lysis of the wild type tumor cell lines (black) and over-expression tumor cell lines (blue) obtained by bioluminescence assay is plotted against multiple E:T ratios. Dots of (m–r) represents three technical replicates (n = 1 experiment), data shown are representative of responses from n = 3 separate donors run independently (Donor S1-S3). Source data are provided as a Source Data file.