Unbiased molecular analysis of T cell receptor expression using template-switch anchored RT-PCR

Curr Protoc Immunol. 2011 Aug:Chapter 10:Unit10.33. doi: 10.1002/0471142735.im1033s94.

Abstract

A detailed knowledge of the principles that guide clonal selection within the memory and effector T cell pools is essential to further our understanding of the factors that influence effective T cell-mediated immunity and has direct implications for the rational design of vaccines and immunotherapies. This unit provides methods for the unbiased quantification and characterization of all expressed T cell receptor (TCR) gene products within any defined T cell population. The approach is based on a template-switch anchored reverse transcription-polymerase chain reaction (RT-PCR) and is optimized for the analysis of antigen-specific T cells isolated directly ex vivo.

MeSH terms

  • Animals
  • Clone Cells
  • Genes, T-Cell Receptor* / genetics
  • Humans
  • Immunity, Cellular / genetics
  • Pathology, Molecular / instrumentation
  • Pathology, Molecular / methods
  • Polymerase Chain Reaction*
  • T-Cell Antigen Receptor Specificity
  • T-Lymphocytes / immunology*
  • Templates, Genetic