title: Transduction with a fiber-modified adenoviral vector is superior to non-viral nucleofection for expressing tumor-associated Ag mucin-1 in human DC.
abstract: 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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Department of Internal Medicine, Division of Hemato-Oncology, University Hospital Maastricht, Maastricht, The Netherlands.
authors.forename: E B M
authors.initials: EB
authors.lastname: van Leeuwen
authors.forename: S
authors.initials: S
authors.lastname: Cloosen
authors.forename: B L M G
authors.initials: BL
authors.lastname: Senden-Gijsbers
authors.forename: W T V
authors.initials: WT
authors.lastname: Germeraad
authors.forename: G M J
authors.initials: GM
authors.lastname: Bos
mesh_terms: D000256:Adenoviridae / Q000235:genetics* / Q000502:physiology; D000911:Antibodies, Monoclonal / Q000276:immunology; D000951:Antigens, Neoplasm / Q000235:genetics / Q000378:metabolism; D049109:Cell Proliferation; D002470:Cell Survival; D002478:Cells, Cultured; D003713:Dendritic Cells / Q000166:cytology / Q000276:immunology* / Q000378:metabolism*; D018274:Electroporation; D005822:Genetic Vectors / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D016753:Interleukin-10 / Q000096:biosynthesis; D018664:Interleukin-12; D007959:Lymphocyte Culture Test, Mixed; D018396:Mucin-1; D009077:Mucins / Q000235:genetics / Q000378:metabolism*; D010641:Phenotype; D014161:Transduction, Genetic; D019562:Viral Load
chemical_list: D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm; C060192:MUC1 protein, human; D018396:Mucin-1; D009077:Mucins; D016753:Interleukin-10; D049452:Green Fluorescent Proteins; D018664:Interleukin-12