
We previously established methods to identify neoantigen-specific TCRs based on patients' PBMCs. In this study, we substituted blood-derived T cells for tumor-infiltrating lymphocytes and used an HLA-matched cell line of antigen-presenting cells to replace autologous dendritic cells. TCRs were isolated from reactive TIL cultures and functionality was tested using TCR- T cells in vitro and in vivo. To exemplify the screening approach, we identified the targeted neoantigen leading to recognition of the minigene construct that stimulated the strongest TIL response. Neoantigen peptides were used to load MHC-tetramers for T cell isolation and a TCR was identified targeting the KIAA1429D1358E mutation. TCR-T cells were activated, exhibited cytotoxicity, and secreted cytokines in a dose-dependent manner,..
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