title: Sensitive detection of human papillomavirus type 16 E7-specific T cells by ELISPOT after multiple in vitro stimulations of CD8+ T cells with peptide-pulsed autologous dendritic cells.
abstract: BACKGROUND
  Cervical cancer is the second most common gynecological cancer amongst women world-wide. Despite optimized protocols, standard treatments still face several disadvantages. Therefore, research aims at the development of immune-based strategies using tumor antigen-loaded dendritic cells for the induction of cellular anti-tumor immunity.
  
  
  RESULTS
  In this study, we used dendritic cells loaded with the HLA-A2-restricted HPV type 16 E711-20 peptide in order to induce an in vitro CD8+ T cell response. For this purpose, peptide-pulsed dendritic cells were co-cultured with autologous CD8+ T cells. After 5 weekly stimulations with peptide-pulsed mature dendritic cells, cultured T cells were analyzed for antigen specificity by an IFN-gamma ELISPOT assay. Using this ELISPOT assay, we were able to detect E7-specific IFN-gamma-secreting CD8+ T cells in 5/5 healthy donors.
  
  
  CONCLUSIONS
  We show that peptide-pulsed mature dendritic cells are able to stimulate a HPV type 16 E7 peptide-specific immune response in vitro. These experiments describe an efficient culture protocol for antigen-specific T cells for use in pre-clinical vaccination research and confirm the need for sensitive T cell assays for detection of tumor-specific immune responses in vitro.
authors.affiliation: Laboratory of Experimental Hematology, Antwerp University, Faculty of Medicine, Antwerp University Hospital, Belgium. nathalie.cools@uza.be
authors.forename: Nathalie
authors.initials: N
authors.lastname: Cools
authors.forename: Peter
authors.initials: P
authors.lastname: Ponsaerts
authors.forename: Marc
authors.initials: M
authors.lastname: Lenjou
authors.forename: Griet
authors.initials: G
authors.lastname: Nijs
authors.forename: Dirk R
authors.initials: DR
authors.lastname: Van Bockstaele
authors.forename: Viggo F I
authors.initials: VF
authors.lastname: Van Tendeloo
authors.forename: Zwi N
authors.initials: ZN
authors.lastname: Berneman
mesh_terms: D000957:Antigens, Viral, Tumor / Q000276:immunology*; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D018920:Coculture Techniques; D003713:Dendritic Cells / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D006801:Humans; D016130:Immunophenotyping; D007371:Interferon-gamma / Q000096:biosynthesis; D020014:K562 Cells; D009842:Oligopeptides / Q000276:immunology; D009856:Oncogene Proteins, Viral / Q000276:immunology*; D050725:Papillomavirus E7 Proteins; D030361:Papillomavirus Infections / Q000276:immunology; D012680:Sensitivity and Specificity; D002583:Uterine Cervical Neoplasms / Q000821:virology
chemical_list: D000957:Antigens, Viral, Tumor; D009842:Oligopeptides; D009856:Oncogene Proteins, Viral; D050725:Papillomavirus E7 Proteins; C059731:oncogene protein E7, Human papillomavirus type 16; D007371:Interferon-gamma