In situ staining using MHC class I tetramers

Curr Protoc Immunol. 2005 Jan:Chapter 17:17.4.1-17.4.9. doi: 10.1002/0471142735.im1704s64.

Abstract

The invention of MHC-tetramer technology to label antigen-specific T cells has lead to a greatly enhanced understanding of T lymphocyte biology. This protocol describes the use of MHC class I tetramers to stain antigen-specific T cells in tissue sections. In situ tetramer staining (IST) can be used to determine the localization, abundance, and phenotype of antigen-specific T cells in native environments and in three-dimensional space. IST is broadly applicable because it can be used to stain essentially any antigen-specific T cell in any tissue for which MHC tetramers are available. IST can be combined with histochemistry and/or immunohistochemistry to permit visualization and characterization of antigen-specific T cells relative to other cell types in stained tissue sections. Thus, IST is a useful and valuable component of MHC-tetramer technology.

MeSH terms

  • Animals
  • Cell Count / methods*
  • Fluorescein-5-isothiocyanate
  • Histocompatibility Antigens Class I* / analysis
  • Histocompatibility Antigens Class I* / metabolism
  • Humans
  • Peptide Fragments* / metabolism
  • Staining and Labeling / methods*
  • T-Lymphocyte Subsets / cytology*
  • T-Lymphocyte Subsets / immunology

Substances

  • Histocompatibility Antigens Class I
  • Peptide Fragments
  • Fluorescein-5-isothiocyanate