Dendritic cell vaccine with mRNA targeted to the proteasome by polyubiquitination

Biochem Biophys Res Commun. 2008 Jun 27;371(2):242-6. doi: 10.1016/j.bbrc.2008.04.034. Epub 2008 Apr 28.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigen Presentation*
  • Antigens, Neoplasm / analysis
  • Antigens, Neoplasm / genetics
  • Antigens, Neoplasm / metabolism*
  • Cancer Vaccines / immunology*
  • Cell Culture Techniques
  • Dendritic Cells / drug effects
  • Dendritic Cells / immunology*
  • Electroporation
  • MART-1 Antigen
  • Mice
  • Mice, Inbred Strains
  • Neoplasm Proteins / analysis
  • Neoplasm Proteins / genetics
  • Neoplasm Proteins / metabolism
  • Proteasome Endopeptidase Complex / metabolism
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • T-Lymphocytes, Cytotoxic / immunology*
  • Transfection
  • Ubiquitination* / genetics

Substances

  • Antigens, Neoplasm
  • Cancer Vaccines
  • MART-1 Antigen
  • Mlana protein, mouse
  • Neoplasm Proteins
  • RNA, Messenger
  • Proteasome Endopeptidase Complex