Marked differences in human melanoma antigen-specific T cell responsiveness after vaccination using a functional microarray

• Published in: PLoS medicine Vol. 2; no. 10; p. e265
• Main Authors: Chen, Daniel S, Soen, Yoav, Stuge, Tor B, Lee, Peter P, Weber, Jeffrey S, Brown, Patrick O, Davis, Mark M
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 01.10.2005; Public Library of Science (PLoS)
• Subjects: Allergy/Immunology; Antibody Formation; Antigens; Antigens, Neoplasm - immunology; Bioengineering; Biotechnology; Cancer Biology; Cancer Vaccines - immunology; CD8-Positive T-Lymphocytes - immunology; Cell Biology; Cytokines; Cytokines - metabolism; Dermatology; General Medicine; Granulocytes; Humans; Immunology; Immunology and Allergy; Infectious Diseases; Interferon; Leukocytes; Lymphocytes; Melanoma; Melanoma - immunology; Melanoma - therapy; Molecular Biology/Structural Biology; Oncology; Peptides; Protein Array Analysis; Systems Biology; T cell receptors; Tumor necrosis factor-TNF; Vaccines; Virology; United States > US; California; CD8-Positive T-Lymphocytes; Antigens, Neoplasm; Cytokines; Cancer Vaccines; Humans; Melanoma; Antibody Formation; Protein Array Analysis
• ISSN: 1549-1676; 1549-1277; 1549-1676
• DOI: 10.1371/journal.pmed.0020265

In contrast to many animal model studies, immunotherapeutic trials in humans suffering from cancer invariably result in a broad range of outcomes, from long-lasting remissions to no discernable effect. In order to study the T cell responses in patients undergoing a melanoma-associated peptide vaccine trial, we have developed a high-throughput method using arrays of peptide-major histocompatibility complexes (pMHC) together with antibodies against secreted factors. T cells were specifically immobilized and activated by binding to particular pMHCs. The antibodies, spotted together with the pMHC, specifically capture cytokines secreted by the T cells. This technique allows rapid, simultaneous isolation and multiparametric functional characterization of antigen-specific T cells present in clinical samples. Analysis of CD8+ lymphocytes from ten melanoma patients after peptide vaccination revealed a diverse set of patient- and antigen-specific profiles of cytokine secretion, indicating surprising differences in their responsiveness. Four out of four patients who showed moderate or greater secretion of both interferon-gamma (IFNgamma) and tumor necrosis factor-alpha (TNFalpha) in response to a gp100 antigen remained free of melanoma recurrence, whereas only two of six patients who showed discordant secretion of IFNgamma and TNFalpha did so. Such multiparametric analysis of T cell antigen specificity and function provides a valuable tool with which to dissect the molecular underpinnings of immune responsiveness and how this information correlates with clinical outcome.