title: Analysis of the T cell response to tumor and viral peptide antigens by an IFNgamma-ELISPOT assay.
abstract: 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.
authors.affiliation: Department of Hematology/Oncology, Medizinische Klinik V, University of Heidelberg, Germany.
authors.forename: C
authors.initials: C
authors.lastname: Scheibenbogen
authors.forename: K H
authors.initials: KH
authors.lastname: Lee
authors.forename: S
authors.initials: S
authors.lastname: Stevanovic
authors.forename: M
authors.initials: M
authors.lastname: Witzens
authors.forename: M
authors.initials: M
authors.lastname: Willhauck
authors.forename: V
authors.initials: V
authors.lastname: Waldmann
authors.forename: H
authors.initials: H
authors.lastname: Naeher
authors.forename: H G
authors.initials: HG
authors.lastname: Rammensee
authors.forename: U
authors.initials: U
authors.lastname: Keilholz
mesh_terms: D000951:Antigens, Neoplasm / Q000276:immunology*; D000956:Antigens, Viral / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D015788:HLA-A1 Antigen / Q000276:immunology; D015789:HLA-A2 Antigen / Q000276:immunology; D006801:Humans; D007371:Interferon-gamma / Q000276:immunology*; D008545:Melanoma / Q000097:blood / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology*
chemical_list: D000951:Antigens, Neoplasm; D000956:Antigens, Viral; D015788:HLA-A1 Antigen; D015789:HLA-A2 Antigen; D007371:Interferon-gamma