Fig. 4. mKRAS TCRαβ gene transfer confers HLA-restricted lytic activity against human tumor cells.

a FACS plot demonstrating p-HLA multimer binding to TCRA3V-, TCRA11V-, and TCRB7R-engineered TCRαβ^null primary CD8⁺ T cells gated on viable/CD3⁺ population. The following p-HLA multimers were used as staining controls: gp17-25/A3, gp17-25/A11, NY60-72/B7 (Supplementary Table 3). b 4 h ⁵¹Cr-release cytotoxicity assay demonstrating TCRA3V, TCRA11V, and TCRB7 cell tumoricidal activity against TMC-expressing (red), mKRAS peptide-pulsed (blue), or WT KRAS peptide-pulsed (black) monoallelic K562 target cells. Percent-specific lysis of triplicates is shown for each data point. Data are presented as mean ± SD. p < 0.05 or p < 0.01 for all E:T ratios ≥0.3:1 comparing TMC-expressing or mKRAS peptide-pulsed to WT KRAS peptide-pulsed K562 targets, respectively; one-way ANOVA followed by post-hoc pair-wise Student’s t-test with multiple comparison adjustment. c, d. 4 h ⁵¹Cr-release cytotoxicity assay demonstrating TCRA3V and TCRA11V tumoricidal activity against cancer cell lines harboring WT KRAS (BxPC-3) or endogenous KRAS G12V mutation either non-modified (open circles, non-HLA matched) or engineered to express the restricting HLA allele (closed circles, HLA-matched). Each color represents a different cell line as identified in the figure legend. No significant cytotoxicity was observed against HLA matched vs non-matched BxPC-3 cell lines. Data are presented as mean values ± SD (n = 3 biologically independent samples). p < 0.05 for all E:T ratio ≥1:1 comparing HLA matched vs non-matched cell lines harboring endogenous KRAS G12V mutation; one-way ANOVA followed by post-hoc pair-wise Student’s t-test with multiple comparison adjustment. Source data are provided as a Source Data file.