Improved detection of melanoma antigen-specific T cells expressing low or high levels of CD8 by HLA-A2 tetramers presenting a Melan-A/Mart-1 peptide analogue

Int J Cancer. 2002 Jan 1;97(1):64-71. doi: 10.1002/ijc.1580.


MHC class I tetramers containing peptide epitopes are sensitive tools for detecting antigen-specific CD8(+) T-cell responses. We demonstrate here that binding of HLA-A2 tetramers to CD8(+) T cells specific for the melanoma-associated antigen Melan-A/MART-1 can be fine-tuned by altering either the bound peptide epitope or residues in the alpha 3 domain of HLA-A2, which is important for CD8 binding. Antigen-specific T cells expressing high levels of CD8 could be detected using HLA-A2 tetramers containing the peptide AAGIGILTV, an epitope which is naturally processed and presented from Melan-A/MART-1. In contrast, low CD8-expressing, antigen-specific T cells could be detected efficiently only by using a mutated HLA-A2 tetramer with an altered CD8 binding site or, less efficiently, using the wild-type HLA-A2 tetramer loaded with the peptide analogue ELAGIGILTV, which is superior in stimulating antigen-specific T-cell responses. Our results suggest ways to optimize the identification and expansion of antigen-specific T cells with different requirements for the costimulatory CD8 molecule in facilitating T-cell receptor binding to peptide variants.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens, Neoplasm / immunology*
  • CD8-Positive T-Lymphocytes / immunology*
  • Cytokines / immunology
  • Cytotoxicity, Immunologic / immunology
  • Epitopes / immunology
  • Flow Cytometry
  • HLA-A2 Antigen / immunology*
  • Humans
  • MART-1 Antigen
  • Melanoma / immunology*
  • Neoplasm Proteins / immunology*
  • Neoplasm Staging
  • Protein Binding / drug effects
  • Protein Binding / immunology
  • Receptors, Antigen, T-Cell / immunology
  • Tumor Cells, Cultured


  • Antigens, Neoplasm
  • Cytokines
  • Epitopes
  • HLA-A2 Antigen
  • MART-1 Antigen
  • MLANA protein, human
  • Neoplasm Proteins
  • Receptors, Antigen, T-Cell