UNIT 6.18 Identification of Human Antigen-Specific T Cells Using MHC Class I and Class II Tetramers

  1. Lori A. Krueger,
  2. C. Thomas Nugent,
  3. Johannes Hampl

Published Online: 1 NOV 2004

DOI: 10.1002/0471142956.cy0618s30

Current Protocols in Cytometry

Current Protocols in Cytometry

How to Cite

Krueger, L. A., Nugent, C. T. and Hampl, J. 2004. Identification of Human Antigen-Specific T Cells Using MHC Class I and Class II Tetramers. Current Protocols in Cytometry. 30:6.18:6.18.1–6.18.12.

Author Information

  1. Beckman Coulter Inc., San Diego, California

Publication History

  1. Published Online: 1 NOV 2004
  2. Published Print: OCT 2004


Major histocompatibility complex (MHC) tetramers typically consist of a fluorophore-streptavidin complex and biotinylated soluble MHC molecules carrying a peptide of interest. Tetramers bind to T cell receptors (TCR) that recognize the MHC molecule/peptide combination with high specificity. Native MHC molecules are expressed as cell-surface glycoproteins capable of binding a variety of peptides generated from the degradation of self and non-self proteins for display to T cells. The human MHC gene locus is highly polymorphic, with >800 class I and >500 class II alleles currently identified. This heterogeneity contributes to the uniqueness of each person's immune system. This unit describes procedures for labeling CD8+ T cells with MHC class I tetramers and CD4+ T cells with MHC class II tetramers. The protocols can be used for detecting and enumerating human antigen-specific T cells. Both CD8+ and CD4+ antigen-specific T cells are rare events and require that sufficient numbers of cells be evaluated. To minimize nonspecific tetramer binding contributed by irrelevant cell populations, a cumulative gating strategy using positive selection and/or exclusion gating is described.


  • MHC tetramers;
  • antigen-specific T cells;
  • flow cytometry