Rapid identification of antigen-specific TCRs for cancer immunotherapyOA

T cell receptors(TCRs)can recognize peptides presented by major histocompatibility complex(MHC)molecules,referred to as HLA in humans,which enables the targeted eradication of tumor cells expressing specific antigens.In recent years,TCR-engineered T cell(TCR-T)cell therapy has demonstrated substantial advancements in clinical trials targeting solid tumors.Notably,in August 2024,the U.S.FDA approved the first TCR-T drug for the treatment of advanced synovial sarcoma,representing a pivotal milestone in the field.For the development of TCR-T therapy,identifying tumor-associated antigen epitopes and high-functional TCRs are critical.Here,we present a comprehensive protocol outlining the process of identification of immunogenic epitopes and the efficient screening of antigen-specific TCRs from HLA transgenic mice.Additionally,the protocol encompasses methodologies for TCR-T cell preparation and their functional evaluation in vitro.These approaches provide a robust framework for advancing the development of tumor-specific TCRs and fostering the clinical translation of TCR-T therapies.