Tracking epitope-specific T cells

Nat Protoc. 2009;4(4):565-81. doi: 10.1038/nprot.2009.9.


The tracking of antigen-specific T cells in vivo is a useful approach for the study of the adaptive immune response. This protocol describes how populations of T cells specific for a given peptide-major histocompatibility complex (pMHC) epitope can be tracked based solely on T-cell receptor (TCR) specificity as opposed to other indirect methods based on function. The methodology involves the adoptive transfer of TCR transgenic T cells with defined epitope specificity into histocompatible mice and the subsequent detection of these cells through the use of congenic or clonotypic markers. Alternatively, endogenous epitope-specific T cells can be tracked directly through the use of pMHC tetramers. Using magnetic bead-based enrichment and advanced multiparameter flow cytometry, populations as small as five epitope-specific T cells can be detected from the peripheral lymphoid organs of a mouse. The adoptive transfer procedure can be completed within 3 h, whereas analysis of epitope-specific cells from mice can be completed within 6 h.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Binding Sites
  • Cell Count / methods
  • Cell Culture Techniques
  • Epitopes, T-Lymphocyte / analysis*
  • Epitopes, T-Lymphocyte / genetics
  • Flow Cytometry / methods*
  • Genetic Markers
  • Magnetics
  • Major Histocompatibility Complex
  • Mice
  • Mice, Transgenic
  • Receptors, Antigen, T-Cell / genetics
  • Receptors, Antigen, T-Cell / metabolism
  • T-Lymphocytes / chemistry
  • T-Lymphocytes / immunology*


  • Epitopes, T-Lymphocyte
  • Genetic Markers
  • Receptors, Antigen, T-Cell