title: A rationally designed tyrosine hydroxylase DNA vaccine induces specific antineuroblastoma immunity.
abstract: Therapeutic vaccination against tumor antigens without induction of autoimmunity remains a major challenge in cancer immunotherapy. Here, we show for the first time effective therapeutic vaccination followed by suppression of established spontaneous neuroblastoma metastases using a tyrosine hydroxylase (TH) DNA minigene vaccine. We identified three novel mouse TH (mTH3) derived peptides with high predicted binding affinity to MHC class I antigen H2-K(k) according to the prediction program SYFPEITHI and computer modeling of epitopes into the MHC class I antigen binding groove. Subsequently, a DNA minigene vaccine was generated based on the expression vector pCMV-F3Ub encoding mutated ubiquitin (Gly(76) to Ala(76)) and mTH3. Prophylactic and therapeutic efficacies of this vaccine were established following oral delivery with attenuated Salmonella typhimurium SL7207. Only mice immunized with mTH3 were free of spontaneous liver metastases. This effect was clearly dependent on ubiquitin and high affinity of the mTH epitopes to MHC class I antigens. Specifically, we showed a crucial role for minigene expression as a stable ubiquitin-Ala(76) fusion peptide for vaccine efficacy. The immune response following the mTH3 DNA minigene vaccination was mediated by CD8(+) T cells as indicated by infiltration of primary tumors and TH-specific cytolytic activity in vitro. Importantly, no cell infiltration was detectable in TH-expressing adrenal medulla, indicating the absence of autoimmunity. In summary, we show effective therapeutic vaccination against neuroblastoma with a novel rationally designed TH minigene vaccine without induction of autoimmunity providing an important baseline for future clinical application of this strategy.
authors.affiliation: Charité-Universitätsmedizin Berlin, Department of Pediatrics, Campus Virchow, Augustenburger Platz 1, 13353 Berlin, Germany.
authors.forename: Nicole
authors.initials: N
authors.lastname: Huebener
authors.forename: Stefan
authors.initials: S
authors.lastname: Fest
authors.forename: Anne
authors.initials: A
authors.lastname: Strandsby
authors.forename: Elke
authors.initials: E
authors.lastname: Michalsky
authors.forename: Robert
authors.initials: R
authors.lastname: Preissner
authors.forename: Yan
authors.initials: Y
authors.lastname: Zeng
authors.forename: Gerhard
authors.initials: G
authors.lastname: Gaedicke
authors.forename: Holger N
authors.initials: HN
authors.lastname: Lode
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000918:Antibody Specificity / Q000276:immunology; D019556:COS Cells; D002522:Chlorocebus aethiops; D003602:Cytotoxicity, Immunologic; D015195:Drug Design; D015395:Histocompatibility Antigens Class I / Q000276:immunology; D007109:Immunity / Q000276:immunology*; D008213:Lymphocyte Activation; D016246:Lymphocytes, Tumor-Infiltrating / Q000276:immunology; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009447:Neuroblastoma / Q000276:immunology* / Q000517:prevention & control*; D010455:Peptides / Q000737:chemistry; D013601:T-Lymphocytes / Q000276:immunology; D014446:Tyrosine 3-Monooxygenase / Q000276:immunology*; D025801:Ubiquitin / Q000378:metabolism; D014611:Vaccination; D019444:Vaccines, DNA / Q000276:immunology*
chemical_list: D015395:Histocompatibility Antigens Class I; D010455:Peptides; D025801:Ubiquitin; D019444:Vaccines, DNA; D014446:Tyrosine 3-Monooxygenase