Extended Data Fig. 7: Validation of antigen-specific TCRs upon TCR cloning.
From: Sensitive identification of neoantigens and cognate TCRs in human solid tumors
a-c, Representative examples of validation of TAA- and neoepitope-specific TCRαβ pairs from patients 1, 2 and 4. a-c, Validation of tumor antigen-specificity after labeling with cognate pMHC multimers of Jurkat cells co-electroporated with TCRα and TCRβ chain RNAs. Dot plots report the concomitant expression of the transgenic TCR and of the mouse TCRβ constant region used as a marker of transfection efficiency. d, For patient 3, tyrosinase508-514-specific TILs were FACS-sorted based on 4-1BB upregulation. Autologous activated primary T cells cloned with TCRαβ pair were co-cultured with autologous CD40-act B cells pulsed with peptide LPEEKQPL. Reactivity was assessed by 4-1BB upregulation. MOCK: control of transfection, neg pair: irrelevant TCRα/β pair, UNP: unpulsed, no antigen.
