title: Marked differences in human melanoma antigen-specific T cell responsiveness after vaccination using a functional microarray.
abstract: BACKGROUND
  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.
  
  
  RESULTS
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Department of Internal Medicine/Division of Oncology, Stanford University, Stanford, California, United States of America.
authors.forename: Daniel S
authors.initials: DS
authors.lastname: Chen
authors.forename: Yoav
authors.initials: Y
authors.lastname: Soen
authors.forename: Tor B
authors.initials: TB
authors.lastname: Stuge
authors.forename: Peter P
authors.initials: PP
authors.lastname: Lee
authors.forename: Jeffrey S
authors.initials: JS
authors.lastname: Weber
authors.forename: Patrick O
authors.initials: PO
authors.lastname: Brown
authors.forename: Mark M
authors.initials: MM
authors.lastname: Davis
mesh_terms: D000917:Antibody Formation; D000951:Antigens, Neoplasm / Q000276:immunology*; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D019496:Cancer Vaccines / Q000276:immunology*; D016207:Cytokines / Q000378:metabolism; D006801:Humans; D008545:Melanoma / Q000276:immunology* / Q000628:therapy; D040081:Protein Array Analysis
chemical_list: D000951:Antigens, Neoplasm; D019496:Cancer Vaccines; D016207:Cytokines