Diverse expansion potential and heterogeneous avidity in tumor-associated antigen-specific T lymphocytes from primary melanoma patients

Eur J Immunol. 2001 Feb;31(2):412-20. doi: 10.1002/1521-4141(200102)31:2<412::aid-immu412>3.0.co;2-4.


While tumor-associated antigen (TAA)-specific CD8(+) T lymphocytes have been detected in metastatic melanoma patients, immune response in early disease phases has not yet been carefully evaluated. We looked for circulating cytotoxic T lymphocytes (CTL) directed against Melan-A / MART1, tyrosinase, gp100 and MAGE-3 antigens in patients with a diagnosis of primary cutaneous melanoma by using fluorescent HLA-A2 tetramers. In five out of six cases high numbers of CD8(+)/tetramer(+) cells could be detected by flow cytometry, and in four patients lymphocyte populations specific for two different melanoma antigens (Melan-A/MART1 and tyrosinase) were contemporaneously present. The TAA-specific cells could represent as much as 1/220 T lymphocytes in the circulating CD8(+) population. When tetramers were used to monitor the in vitro expansion of TAA-specific CTL precursors upon antigen-specific stimulation, a diverse expansion potential was evidenced in CTL from the different donors and, more strikingly, in CTL specific for the different TAA. Melan-A/MART1-specific CTL clones derived from two patients exhibited a broad range of avidity. Only the highest avidity clones, representing about 50 % of the cases analyzed, were tumor specific. By correlating tetramer staining with clone avidity, we found that tetramer fluorescence intensity could represent a good indicator of TCR affinity, but not of overall clone avidity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Antigens, Neoplasm / immunology*
  • Female
  • Fluorescence
  • HLA-A2 Antigen / chemistry
  • HLA-A2 Antigen / metabolism
  • Humans
  • MART-1 Antigen
  • Male
  • Melanoma / immunology*
  • Membrane Glycoproteins / immunology
  • Middle Aged
  • Monophenol Monooxygenase / immunology
  • Neoplasm Proteins / immunology
  • Receptors, Antigen, T-Cell / metabolism
  • T-Lymphocytes, Cytotoxic / immunology*
  • gp100 Melanoma Antigen


  • Antigens, Neoplasm
  • HLA-A2 Antigen
  • MAGEA3 protein, human
  • MART-1 Antigen
  • MLANA protein, human
  • Membrane Glycoproteins
  • Neoplasm Proteins
  • PMEL protein, human
  • Receptors, Antigen, T-Cell
  • gp100 Melanoma Antigen
  • Monophenol Monooxygenase