title: Single-Cell Analysis of T-Cell Receptor αβ Repertoire.
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.
authors.affiliation: Department of Immunology, St. Jude Children's Research Hospital, 262 Danny Thomas Place/MS 351‬, Memphis, TN, 38105, USA. pradyot.dash@stjude.org.
authors.forename: Pradyot
authors.initials: P
authors.lastname: Dash
authors.affiliation: Department of Immunology, St. Jude Children's Research Hospital, 262 Danny Thomas Place/MS 351‬, Memphis, TN, 38105, USA.|Center of Excellence in Geriatric Medicine, Newton Medical Center, Sparta, NJ, USA.
authors.forename: George C
authors.initials: GC
authors.lastname: Wang
authors.affiliation: Department of Immunology, St. Jude Children's Research Hospital, 262 Danny Thomas Place/MS 351‬, Memphis, TN, 38105, USA. paul.thomas@stjude.org.
authors.forename: Paul G
authors.initials: PG
authors.lastname: Thomas
mesh_terms: D005434:Flow Cytometry / Q000379:methods; D006801:Humans; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D059010:Single-Cell Analysis / Q000379:methods*; D013601:T-Lymphocytes / Q000378:metabolism*
chemical_list: D016693:Receptors, Antigen, T-Cell, alpha-beta
keywords: Clonotype; Complementarity-determining region 3; Epitope; Paired analysis; Single-cell analysis; T-cell receptor repertoire; TCR alpha; TCR beta; TCR diversity; Unbiased repertoire