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Antigen identification for HLA class I– and HLA class II–restricted T cell receptors using cytokine-capturing antigen-presenting cells

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Science Immunology  22 Jan 2021:
Vol. 6, Issue 55, eabf4001
DOI: 10.1126/sciimmunol.abf4001

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Finding TCR epitopes in peptide haystacks

Identification of the peptide-MHC ligands recognized by a specific T cell receptor (TCR) remains a formidable bottleneck for T cell biology. Lee and Meyerson developed an epitope discovery platform based on introduction of DNA-encoded peptide pools into immortalized antigen-presenting cells (APCs) engineered to express defined class I or II MHC molecules. When αβ T cells of interest were activated by a cognate peptide-MHC complex, secreted cytokines were captured by anti-cytokine antibodies tethered to the APC surface. Positive selection for cytokine-displaying APCs followed by next-generation DNA sequencing revealed the peptide epitope seen by the TCR. This approach enabled a proof-of-principle experiment that defined several previously unknown CD4+ and CD8+ T cell epitopes from cytomegalovirus recognized by “public” TCRs shared among multiple CMV seropositive donors.

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