Transduction with a fiber-modified adenoviral vector is superior to non-viral nucleofection for expressing tumor-associated Ag mucin-1 in human DC

Cytotherapy. 2006;8(1):36-46. doi: 10.1080/14653240500508166.


Background: DC-presenting tumor Ag are currently being developed to be used as a vaccine in human cancer immunotherapy. To increase the chances for successful therapy it is important to deliver full-length tumor Ag instead of loading single peptides. Methodologically, several recombinant DNA delivery techniques have been used.

Methods: In this study we compared nucleofection, an optimized form of electroporation, and adenoviral transduction regarding their efficiency to transduce human monocyte-derived (Mo-) DC in vitro. Expression of the tumor-associated Ag mucin-1 (MUC1) after adenoviral transduction (rAd5Fib35-MUC1) was determined using two MAb.

Results: We showed that the viability of cells and percentage of green fluorescent protein (GFP)-positive cells after transduction with a fiber-modified adenoviral vector (rAd5F35-GFP) was much higher than after nucleofection. Furthermore, phenotype and function of DC were not impaired by infection with adenovirus particles. Cells matured normally; up-regulation of CD40, CD80, CD83, CD86 and HLA-DR was not affected by adenoviral transduction. The capacity to stimulate naive T-cell proliferation was preserved and no change in IL-10 production was observed. Production of IL-12 increased up to 500-fold upon adenoviral transduction, considered to contribute positively to an anti-tumor immune response. Non-transduced mature DC expressed low levels of endogenous MUC1. After transduction with the rAd5F35-MUC1 adenoviral vector, a 100-fold increase in MUC1 expression by DC was observed.

Discussion: The use of the fiber-modified adenoviral vector presented here may therefore be favorable compared with non-viral gene delivery systems for DC that will be used in cancer immunotherapy.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenoviridae / genetics*
  • Adenoviridae / physiology
  • Antibodies, Monoclonal / immunology
  • Antigens, Neoplasm / genetics
  • Antigens, Neoplasm / metabolism
  • Cell Proliferation
  • Cell Survival
  • Cells, Cultured
  • Dendritic Cells / cytology
  • Dendritic Cells / immunology*
  • Dendritic Cells / metabolism*
  • Electroporation
  • Genetic Vectors / genetics
  • Green Fluorescent Proteins / metabolism
  • Humans
  • Interleukin-10 / biosynthesis
  • Interleukin-12
  • Lymphocyte Culture Test, Mixed
  • Mucin-1
  • Mucins / genetics
  • Mucins / metabolism*
  • Phenotype
  • Transduction, Genetic*
  • Viral Load


  • Antibodies, Monoclonal
  • Antigens, Neoplasm
  • MUC1 protein, human
  • Mucin-1
  • Mucins
  • Interleukin-10
  • Green Fluorescent Proteins
  • Interleukin-12