title: Dendritic cell vaccine with mRNA targeted to the proteasome by polyubiquitination.
abstract: Dendritic cells (DCs) transfected with mRNA encoding tumor-associated antigens (TAAs) can induce tumor-specific T-cell responses. To potentiate this, we transfected mature DCs (mDCs) with mRNA encoding TAA targeted to the proteasome. DCs were generated from bone marrow cells by culture with 20 ng/ml GM-CSF and maturation with 1 microg/ml LPS. These mDCs were then electroporated with 10 microg of mRNA. Antigen presentation after electroporation with in vitro transcribed mRNA was compared with mRNA from a construct of the TAA preceded by ubiquitin. Proteasomal targeting of mRNA encoding cotranslationally ubiquitinated antigen was found to enhance intracellular degradation of target protein, and result in more efficient priming and expansion of TAA-specific CD8(+) T-cells. We therefore suggest that RNA-transfected DC vaccine efficacy could be improved by the use of mRNA targeted to the proteasome.
authors.affiliation: Department of Immunotherapeutics (Medinet), Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-Ku, Tokyo 113-8655, Japan.
authors.forename: Akihiro
authors.initials: A
authors.lastname: Hosoi
authors.forename: Yayoi
authors.initials: Y
authors.lastname: Takeda
authors.forename: Kazuko
authors.initials: K
authors.lastname: Sakuta
authors.forename: Satoshi
authors.initials: S
authors.lastname: Ueha
authors.forename: Makoto
authors.initials: M
authors.lastname: Kurachi
authors.forename: Kiminori
authors.initials: K
authors.lastname: Kimura
authors.forename: Ryuji
authors.initials: R
authors.lastname: Maekawa
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Kakimi
mesh_terms: D000818:Animals; D017951:Antigen Presentation; D000951:Antigens, Neoplasm / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D019496:Cancer Vaccines / Q000276:immunology*; D018929:Cell Culture Techniques; D003713:Dendritic Cells / Q000187:drug effects / Q000276:immunology*; D018274:Electroporation; D058965:MART-1 Antigen; D051379:Mice; D008815:Mice, Inbred Strains; D009363:Neoplasm Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D014162:Transfection; D054875:Ubiquitination / Q000235:genetics
chemical_list: D000951:Antigens, Neoplasm; D019496:Cancer Vaccines; D058965:MART-1 Antigen; C548668:Mlana protein, mouse; D009363:Neoplasm Proteins; D012333:RNA, Messenger; D046988:Proteasome Endopeptidase Complex