Optimization of dendritic cell maturation and gene transfer by recombinant adenovirus Journal Article

Authors: Miller, G.; Lahrs, S.; Shah, A. B.; DeMatteo, R. P.
Article Title: Optimization of dendritic cell maturation and gene transfer by recombinant adenovirus
Abstract: Dendritic cells (DC) have vast potential for immunotherapy. Transferring therapeutic genes to DC may enhance their inherent T cell-stimulatory capacity. Recombinant adenovirus is the most efficient vehicle for DC gene transfer and can alone mature DC. We sought to define the parameters of adenovirus infection of murine bone marrow-derived DC (BMDC) and the concomitant impact on BMDC maturation. The efficiency of adenoviral gene transfer to DC depended on the mouse strain, the organ source of DC, and the level of DC maturation. C57BL/6 BMDC consistently had higher transgene expression than BALB/c DC. While BMDC had considerable GFP expression after AdGFP infection, adenovirus was relatively ineffective in accomplishing transgene expression in freshly isolated hepatic or splenic DC. BMDC that were relatively immature because of a shorter duration of culture had higher transgene expression after infection. Nevertheless, pretreatment of DC with exogenous stimulants such as LPS or TNF-α resulted in higher transgene expression. Maturation of BMDC depended only on virus entry but not viral gene or transgene expression. Therefore, DC maturation was disproportionately high compared to the percentage of DC that actually expressed the adenoviral transgene. Maturation by adenovirus was only seen in BMDC, but not in liver or splenic DC, and was more pronounced in DC from later in culture (day 12 versus day 6). There was a dose-response relationship, up to a threshold dose, between adenovirus infection and both DC maturation and enhancement of DC activation of antigen-specific T cells. Our findings underscore the importance of optimizing gene transfer to DC in designing strategies for immunotherapy.
Keywords: controlled study; nonhuman; flow cytometry; t-lymphocytes; animal cell; mouse; animals; mice; microscopy, electron; cell division; gene expression; cell maturation; spleen; microscopy, confocal; dendritic cell; green fluorescent protein; dose-response relationship, drug; mice, inbred balb c; mice, inbred c57bl; time factors; gene transfer; viral gene delivery system; luminescent proteins; liver; dendritic cells; immunotherapy; recombinant proteins; cell isolation; transgene; green fluorescent proteins; immunophenotyping; immunostimulation; virus infection; bone marrow cell; virus recombinant; mouse strain; liver cell; t lymphocyte activation; transgenes; adenoviridae; spleen cell; gene transfer techniques; adenovirus; male; priority journal; article; virus adsorption
Journal Title: Cancer Immunology, Immunotherapy
Volume: 52
Issue: 6
ISSN: 0340-7004
Publisher: Springer  
Date Published: 2003-06-01
Start Page: 347
End Page: 358
Language: English
PUBMED: 12739067
PROVIDER: scopus
DOI: 10.1007/s00262-003-0379-6
Notes: Export Date: 12 September 2014 -- Source: Scopus
Citation Impact
MSK Authors
  1. George Miller
    18 Miller
  2. Ronald P DeMatteo
    635 DeMatteo
  3. Alaap Shah
    20 Shah
  4. Svenja   Lahrs
    9 Lahrs