Fig. 1.

G12D mutation is critical for presentation and T cell recognition of KRAS-G12D neoantigens. (A) Strategy to study TIL TCRs. (B and C) HLA-I stabilization on TAP-deficient 221-C*08:02-ICP47 cells incubated overnight at 26 °C with 100 μM WT and G12D KRAS 9- and 10-mer and control peptide YVD (YVDEHGTRL). NP, no peptide. A representative experiment is shown in B and data from five or six independent experiments are summarized in C. Statistical significance was assessed by one-way ANOVA with Tukey’s multiple comparison test (****P < 0.0001). (D and E) Frequency of TCR⁺ Jurkat T cells expressing CD69 after incubation with 221-C*08:02-ICP47 cells loaded with WT and G12D KRAS 9- and 10-mer, as measured by flow cytometry. Peptides were tested from 1,000 to 0.01 nM. A representative experiment is the data shown in D with 10 nM peptide and four to six independent experiments are summarized in E with mean and SEM values. MdFI = median fluorescence intensity.