Analysis of the T cell response to tumor and viral peptide antigens by an IFNgamma-ELISPOT assay

Int J Cancer. 1997 Jun 11;71(6):932-6. doi: 10.1002/(sici)1097-0215(19970611)71:6<932::aid-ijc3>;2-z.


We have established a sensitive ELISPOT assay measuring interferon gamma (IFN gamma) release on a single-cell basis to detect influenza peptide-specific CD8+ T cells in uncultured peripheral blood mononuclear cells (PBMC). Using this method, we studied the T cell response to HLA-A1 and HLA-A2.1 binding peptide epitopes derived from the MAGE-1 and MAGE-3 proteins, from the melanoma-associated antigens tyrosinase, Melan-A/MART-1 and gp100, and from influenza proteins in stage IV melanoma patients and healthy controls. In 18 of 24 HLA-A2-positive donors (75%), but only in 9 of 25 HLA-A2-positive melanoma patients (36%) T cells reactive with the influenza matrix peptide were demonstrated (p = 0.007). T cells responding to one or several of the melanoma-associated peptides were detected in 5 of 25 HLA-A2-positive patients with metastatic melanoma. Four of these 5 patients had been treated with interleukin-2- and IFN alpha-containing therapy. Two of the 24 healthy donors had T cells reactive with the MART-1 27-35 peptide. No reactivity with the HLA-A1-binding peptides from MAGE-1 or MAGE-3 was detected in any of the HLA-A1-positive healthy controls or melanoma patients. These results show that the IFN gamma-ELISPOT assay is suitable to determine quantitatively T cells reactive with melanoma-associated and influenza peptide epitopes in uncultured PBMC. The failure to detect T cells responding to influenza in many melanoma patients with progressive disease may indicate an impairment of their T cell function.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens, Neoplasm / immunology*
  • Antigens, Viral / immunology*
  • Enzyme-Linked Immunosorbent Assay
  • HLA-A1 Antigen / immunology
  • HLA-A2 Antigen / immunology
  • Humans
  • Interferon-gamma / immunology*
  • Melanoma / blood
  • Melanoma / immunology
  • T-Lymphocytes / immunology*


  • Antigens, Neoplasm
  • Antigens, Viral
  • HLA-A1 Antigen
  • HLA-A2 Antigen
  • Interferon-gamma