Single-cell analysis of T-cell receptor αβ repertoire

Pradyot Dash, George C. Wang, Paul G. Thomas

Research output: Chapter in Book/Report/Conference proceedingChapter

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.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages181-197
Number of pages17
DOIs
StatePublished - Jan 1 2015

Publication series

NameMethods in Molecular Biology
Volume1343
ISSN (Print)1064-3745

Keywords

  • Clonotype
  • Complementarity-determining region 3
  • Epitope
  • Paired analysis
  • Single-cell analysis
  • T-cell receptor repertoire
  • TCR alpha
  • TCR beta
  • TCR diversity
  • Unbiased repertoire

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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  • Cite this

    Dash, P., Wang, G. C., & Thomas, P. G. (2015). Single-cell analysis of T-cell receptor αβ repertoire. In Methods in Molecular Biology (pp. 181-197). (Methods in Molecular Biology; Vol. 1343). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-2963-4_15