Dendritic cell vaccine with mRNA targeted to the proteasome by polyubiquitination.
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.
Hosoi
Akihiro
A
Department of Immunotherapeutics (Medinet), Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-Ku, Tokyo 113-8655, Japan.
Takeda
Yayoi
Y
Sakuta
Kazuko
K
Ueha
Satoshi
S
Kurachi
Makoto
M
Kimura
Kiminori
K
Maekawa
Ryuji
R
Kakimi
Kazuhiro
K
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
D000951:Antigens, Neoplasm; D019496:Cancer Vaccines; D058965:MART-1 Antigen; C548668:Mlana protein, mouse; D009363:Neoplasm Proteins; D012333:RNA, Messenger; D046988:Proteasome Endopeptidase Complex