title: Cotransfection of dendritic cells with RNA coding for HER-2/neu and 4-1BBL increases the induction of tumor antigen specific cytotoxic T lymphocytes.
abstract: Ribonucleic acid (RNA) transfection of dendritic cells (DCs) was shown to be highly efficient in eliciting CD8+ and CD4+ T-cell responses. We analyzed whether electroporation of DCs with RNA coding for a tumor-associated antigen (TAA) would elicit antigen-specific effector cytotoxic T lymphocyte (CTL) responses and whether these responses could be modulated by cotransfection with a second specific synthetic RNA. Therefore in vitro generated human monocyte-derived DCs were electroporated with in vitro transcribed RNA (in vitro transcript, IVT) encoding the TAA HER-2/neu. Additionally, these cells were cotransfected with IVT coding for human 4-1BBL. Transfection of DCs with 4-1BBL-IVT did not alter their typical phenotype. However, it increased the expression of the costimulatory molecules CD80 and CD40. Coadministration of HER-2/neu- and 4-1BBL-IVT resulted in an increased specific lysis of target cells by the in vitro induced CTL lines, indicating that 4-1BBL enhances their ability to elicit primary CTL responses. Interestingly, transfection of DCs with 4-1BBL-IVT did not augment their capacity to stimulate allogeneic lymphocyte responses. The here established approach of cotransfection of DCs with tumor-RNA and a second specific IVT could improve and optimize the in vitro manipulation of DCs for the induction of antigen-specific CTL responses.
authors.affiliation: Department of Internal Medicine II, Hematology, Oncology and Immunology, University of Tübingen, Tübingen, Germany.
authors.forename: Frank
authors.initials: F
authors.lastname: Grünebach
authors.forename: Katrin
authors.initials: K
authors.lastname: Kayser
authors.forename: Markus M
authors.initials: MM
authors.lastname: Weck
authors.forename: Martin R
authors.initials: MR
authors.lastname: Müller
authors.forename: Silke
authors.initials: S
authors.lastname: Appel
authors.forename: Peter
authors.initials: P
authors.lastname: Brossart
mesh_terms: D053330:4-1BB Ligand; D000951:Antigens, Neoplasm / Q000235:genetics* / Q000276:immunology; D018122:B7-1 Antigen / Q000032:analysis / Q000378:metabolism; D019013:CD40 Antigens / Q000032:analysis / Q000378:metabolism; D003713:Dendritic Cells / Q000276:immunology* / Q000378:metabolism; D015316:Genetic Therapy; D006801:Humans; D016056:Immunodominant Epitopes / Q000235:genetics / Q000276:immunology; D007167:Immunotherapy / Q000379:methods; D009369:Neoplasms / Q000276:immunology* / Q000628:therapy; D012333:RNA, Messenger / Q000235:genetics; D012334:RNA, Neoplasm / Q000235:genetics*; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000235:genetics* / Q000378:metabolism; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology* / Q000378:metabolism; D014162:Transfection; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics* / Q000276:immunology; D015854:Up-Regulation
chemical_list: D053330:4-1BB Ligand; D000951:Antigens, Neoplasm; D018122:B7-1 Antigen; D019013:CD40 Antigens; D016056:Immunodominant Epitopes; D012333:RNA, Messenger; D012334:RNA, Neoplasm; D018719:Erb-b2 Receptor Tyrosine Kinases; D014409:Tumor Necrosis Factor-alpha; C505974:TNFSF9 protein, human