Specific T cell receptor targeting cytomegalovirus antigen and application thereof
A technology of cytomegalovirus and cell receptors, which is applied in the field of specific T cell receptors targeting cytomegalovirus antigens, can solve the problems of missing TCR pairing information and complicated screening procedures, and achieve the effect of simple and easy-to-operate preparation methods
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[0060] In this case, antigen-specific cytotoxic T lymphocytes were induced and expanded in vitro based on HLA-A*0201-typed antigen CMV pp65 antigen NLV epitope polypeptide NLVPMVATV, and positive T cells were sorted by single-cell sequencing and MHC tetramer According to the TCR pairing information, TCR was screened according to the clone sequence frequency, TCR-T was constructed, functional verification was carried out in vitro, and the function of TCR-T was evaluated to finally determine the specific T cell receptor in this case. The details are as follows:
[0061] 1. Induction and functional detection of CMV pp65 NLV-specific cytotoxic T lymphocytes
[0062] 1.1 Induction of CMV pp65 NLV-specific cytotoxic T lymphocytes
[0063] 50 mL of peripheral blood from healthy people with HLA-A*0201 typing was collected, and peripheral blood mononuclear cells (PBMC) were obtained by Ficoll density gradient centrifugation. From the isolated PBMC, CD14 was sorted using anti-CD14 ant...
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