Analysis of the paired TCR α- and β-chains of single human T cells

PLoS One. 2012;7(5):e37338. doi: 10.1371/journal.pone.0037338. Epub 2012 May 23.


Analysis of the paired i.e. matching TCR α- and β-chain rearrangements of single human T cells is required for a precise investigation of clonal diversity, tissue distribution and specificity of protective and pathologic T-cell mediated immune responses. Here we describe a multiplex RT-PCR based technology, which for the first time allows for an unbiased analysis of the complete sequences of both α- and β-chains of TCR from single T cells. We validated our technology by the analysis of the pathologic T-cell infiltrates from tissue lesions of two T-cell mediated autoimmune diseases, psoriasis vulgaris (PV) and multiple sclerosis (MS). In both disorders we could detect various T cell clones as defined by multiple T cells with identical α- and β-chain rearrangements distributed across the tissue lesions. In PV, single cell TCR analysis of lesional T cells identified clonal CD8(+) T cell expansions that predominated in the epidermis of psoriatic plaques. An MS brain lesion contained two dominant CD8(+) T-cell clones that extended over the white and grey matter and meninges. In both diseases several clonally expanded T cells carried dual TCRs composed of one Vβ and two different Vα-chain rearrangements. These results show that our technology is an efficient instrument to analyse αβ-T cell responses with single cell resolution in man. It should facilitate essential new insights into the mechanisms of protective and pathologic immunity in many human T-cell mediated conditions and allow for resurrecting functional TCRs from any αβ-T cell of choice that can be used for investigating their specificity.

Publication types

  • Research Support, Non-U.S. Gov't
  • Validation Study

MeSH terms

  • Antigen-Presenting Cells / immunology
  • DNA Primers / genetics
  • Flow Cytometry
  • Genes, T-Cell Receptor alpha / genetics*
  • Genes, T-Cell Receptor beta / genetics*
  • Humans
  • Multiple Sclerosis / immunology
  • Multiplex Polymerase Chain Reaction / methods*
  • Psoriasis / immunology
  • Reverse Transcriptase Polymerase Chain Reaction / methods*
  • Skin / immunology
  • T-Lymphocytes / metabolism*


  • DNA Primers