Multiplex MHC Class I Tetramer Combined with Intranuclear Staining by Mass Cytometry

Methods Mol Biol. 2019:1989:147-158. doi: 10.1007/978-1-4939-9454-0_11.

Abstract

Antigen-specific CD8+ T cells play a crucial role in the host protective immune response against viruses, tumors, and other diseases. Major histocompatibility complex (MHC) class I tetramers allow for a direct detection of such antigen-specific CD8+ T cells. Using mass cytometry together with multiplex MHC class I tetramer staining, we are able to screen more than 1000 different antigen candidates simultaneously across tissues in health and disease, while retaining the possibility to deliver an in-depth characterization of antigen-specific CD8+ T cells and associated phenotypes. Here we describe the method for a MHC class I tetramer multiplexing approach together with intracellular antibody staining for a parallel phenotypic cell characterization using mass cytometry in human specimens.

Keywords: Antigen-specific T cell; MHC class I tetramer; Mass cytometry; Multiplex tetramer staining.

MeSH terms

  • CD8-Positive T-Lymphocytes / immunology*
  • Cell Nucleus / immunology
  • Cell Nucleus / metabolism*
  • Flow Cytometry / methods*
  • Histocompatibility Antigens Class I / analysis*
  • Histocompatibility Antigens Class I / immunology
  • Histocompatibility Antigens Class I / metabolism
  • Humans
  • Mass Spectrometry / methods*
  • Neoplasms / immunology*
  • Neoplasms / metabolism
  • Single-Cell Analysis / methods
  • Staining and Labeling / methods*

Substances

  • Histocompatibility Antigens Class I