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

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Abstract

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.

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Figure 1: Combinatorial pMHC tetramer staining.
Figure 2: Specificity of combinatorial pMHC tetramer staining.

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Acknowledgements

We thank K. Adachi, M. Kuhns, Y.-H. Chien, G. Nolan and D. Furman for helpful discussions. This work was supported by National Institutes of Health grant U19AI057229, a Bill and Melinda Gates Foundation Grand Challenges Explorations grant and The Howard Hughes Medical Institute. E.W.N. is supported by The American Cancer Society, California Division, Steven Stanley and Edward Albert Bielfelt postdoctoral fellowship. L.O.K. is supported by a National Science Foundation graduate research fellowship. W.Y. is supported by a Damon Runyon postdoctoral fellowship.

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Correspondence to Mark M Davis.

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Supplementary Figures 1–7 and Supplementary Tables 1–2 (PDF 1079 kb)

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Newell, E., Klein, L., Yu, W. et al. Simultaneous detection of many T-cell specificities using combinatorial tetramer staining. Nat Methods 6, 497–499 (2009). https://doi.org/10.1038/nmeth.1344

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