Extended Data Fig. 3: The CTNNB1-S37F-reactive HLA-A2- and HLA-A24- restricted TCRs recognize mutant but not the corresponding WT peptides.
From: TCR-engineered T cells targeting a shared β-catenin mutation eradicate solid tumors
a, Gating strategy used in Fig. 2a, to identify and sort CD8+ T cells staining double positive for dual-colored pMHC-multimers. b, Gating strategy used in Fig. 2b, c, to identify TCR-transduced cells. Pre-gating was performed as in Extended Data Fig. 3a (three left plots) and show viable CD8+ cells staining positively with anti-mouse TCRβ (middle plot), or with APC- and PE-labeled pMHC-multimers (right plot). c, Activation of CD8+ CTNNB1-S37F TCR-T cells following co-incubation with target cells expressing the relevant HLA in presence or absence of additional loading with the cognate CTNNB1-S37F or CTNNB1-WT peptide for the relevant TCR (1 µM). d, CTNNB1-S37F TCR and control TCR1G4 or TCRHIV T cells were co-incubated with target cells expressing the relevant HLA and pulsed with indicated concentrations of cognate peptide. Activation was measured as percentage of CD137+ cells and the mean EC50 values of the indicated TCRs were calculated. Data are pooled from n = 3 independent experiments and each datapoint represents mean ± s.e.m. of n = 3 different PBMC donors with n = 2 technical replicates. e, Related to Fig. 2e, flow plots showing activation of CD8+ CTNNB1-S37F TCR-T cells following co-incubation with Mel888A24, Hutu80A2, Mel888A24+A2 or Hutu80A2+A24 ± additional loading with the cognate CTNNB1-S37F peptide (100 nM). Data shown for one PBMC donor is representative of n = 2 donors in one experiment.
