Extended Data Fig. 4: The majority of αβILTCKs recognize tumor-associated antigens presented in the context of classical MHC-I.
From: Programme of self-reactive innate-like T cell-mediated cancer immunity
a, A representative flow plot showing the expression of classical MHC-I (H2-Kb and H2-Db) on PyMT cancer cells, defined as CD45–CD31–EpCAM+ from mice of indicated genotypes. Data are representative of two independent experiments. b, Frequency of GFP+ cells among CD8+ reporter cell line expressing indicated TCRs 24 h after co-culturing with primary PyMT cancer cells doubly deficient for H2-Kb and H2-Db. Data are pooled from three independent experiments. c, Flow cytometric analysis showing the frequency of GFP+ cells among CD8+ and CD8– reporter cell line expressing the indicated TCRs 24 h after co-culturing with primary PyMT cancer cells lacking β2m. Data are representative of two independent experiments. d, Frequency of TCRβ+CD8α+ cells among CD45+ tumor-infiltrating cells in mice lacking classical MHC-I (H2-K1–/–H2-D1–/–, n = 3), both classical and non-classical MHC-I (B2m–/–, n = 3) or control wild-type mice (n = 7). The profiles of PD-1 and NK1.1 expression by TCRβ+CD8α+ are shown in the bottom panel. e, Statistical analysis of frequency of PD-1+ T cells and αβILTCKs among tumor-infiltrating CD45+ cells in mice of indicated genotypes. f, Flow cytometric analysis showing H-2Db expression on control, TAP1- or β2m-deficient PyMT cell lines generated by Cas9-mediated genome editing with small guide RNA (sgRNA). g, Expression of GFP in reporter cell lines expressing the indicated TCRs 24 h after co-culture with control or TAP1-deficient PyMT cells lines in the presence of absence of SIINFEKL peptide. h, Frequency of GFP+ cells among TCR-expressing reporter cells after co-culture with PyMT cell lines of indicated genotypes with or without SIINFEKL peptide. Data are pooled from three independent experiments. All statistical data are shown as mean ± S.D (biologically independent mice in a, c–e and independent samples in b, f–h). **P < 0.01; ***P < 0.001 and n.s.: not significant.
