title: Preclinical full-scale evaluation of dendritic cells transfected with autologous tumor-mRNA for melanoma vaccination.
abstract: Most cancer vaccines to date have made use of common tumor antigens or allogenic cancer cell lines. The majority of tumor antigens may, however, be unique patient-specific antigens. Dendritic cells (DCs) are the most potent antigen-presenting cells known. The present report is a full-scale preclinical evaluation of autologous DCs transfected with autologous tumor-mRNA (tDCs) for vaccination in malignant melanoma. By using autologous tumor-mRNA, we intend to make the DCs present a broad spectrum of tumor-associated antigens relevant to each individual patient. Previously, we have described effective methods for mRNA-transfection into DCs by square-wave electroporation and for generating large numbers of DCs. Here, we demonstrate the ability of tDCs, made under full-scale vaccine conditions, to generate in vitro T-cell responses specific for antigens encoded by the transfected tumor-mRNA. T-cell proliferation assays demonstrated tDC-specific responses for all six patients tested. Responses were further studied by IFNgamma ELISPOT and Bioplex cytokine assays (two patients) and by experiments on isolated CD4(+) and CD8(+) T cells, including HLA-blockage (one patient). Moreover, we describe the results of extensive tumor-RNA analysis using Agilent Bioanalyser, a method that we have implemented in the clinical protocol. Based on this preclinical evaluation, a vaccine trial has been started.
authors.affiliation: Section for Immunotherapy, Department of Immunology, Cancer Research Institute, The Norwegian Radium Hospital, University of Oslo, 0310 Oslo, Norway. j.a.kyte@klinmed.uoi.no
authors.forename: Jon A
authors.initials: JA
authors.lastname: Kyte
authors.forename: Gunnar
authors.initials: G
authors.lastname: Kvalheim
authors.forename: Steinar
authors.initials: S
authors.lastname: Aamdal
authors.forename: Stein
authors.initials: S
authors.lastname: Saebøe-Larssen
authors.forename: Gustav
authors.initials: G
authors.lastname: Gaudernack
mesh_terms: D000328:Adult; D000368:Aged; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D019496:Cancer Vaccines / Q000235:genetics / Q000276:immunology*; D003713:Dendritic Cells / Q000276:immunology*; D006801:Humans; D007371:Interferon-gamma / Q000378:metabolism; D008297:Male; D008545:Melanoma / Q000188:drug therapy / Q000276:immunology*; D008875:Middle Aged; D012333:RNA, Messenger / Q000235:genetics*; D012334:RNA, Neoplasm / Q000235:genetics*; D014162:Transfection
chemical_list: D019496:Cancer Vaccines; D012333:RNA, Messenger; D012334:RNA, Neoplasm; D007371:Interferon-gamma