Abstract
The unbiased, paired analysis of T-cell receptor (TCR) α- and β-chain usage at the single-cell level provides a valuable window of understanding into the TCR repertoire and the nature of the immune response. Earlier technologies for TCR repertoire analysis were often limited to examining TCR complementarity-determining region 3 (CDR3) β expression or required in vitro cloning procedures that can artificially skew the TCR repertoire from its in vivo state. We describe here a direct ex vivo, single-cell-based strategy for the clonotypic analysis of TCRαβ repertoires that utilizes multiplexed panels of TCRα and TCRβ-specific primers in a nested PCR to amplify expressed transcripts from individual, epitope-specific T cells. This strategy yields the paired TCRαβ sequences of any given population of αβ T cells of interest.
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Acknowledgments
This work was supported by NIH—National Institute of Allergy and Infectious Diseases grants AI077714 and AI107625, the American Syrian Lebanese Associated Charities, NIH—National Institute on Aging grant AG033113, Atlantic Philanthropies, American Geriatrics Society, the John A. Hartford Foundation, and the Association of Subspecialty Professors.
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Dash, P., Wang, G.C., Thomas, P.G. (2015). Single-Cell Analysis of T-Cell Receptor αβ Repertoire. In: Shaw, A. (eds) Immunosenescence. Methods in Molecular Biology, vol 1343. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2963-4_15
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DOI: https://doi.org/10.1007/978-1-4939-2963-4_15
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2962-7
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