Simultaneous detection of many T-cell specificities using combinatorial tetramer staining

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Abstract
Combinations of fluorescently labeled peptide–major histocompatability (pMHC) tetramers are used to simultaneously detect T cells with multiple antigen specificities from human blood samples. Also in this issue, Hadrup et al. present a very similar combinatorial encoding approach. The direct detection of antigen-specific T cells using tetramers of soluble peptide–major histocompatibilty complex (pMHC) molecules is widely used in both basic and clinical immunology. However, the number of specificities that can be assessed simultaneously has been a major limitation. Here we describe and validate a method using combinations of fluorescent pMHC tetramers to simultaneously detect and enrich for many (≥15) T-cell specificities in a single human blood sample.