Efficient gene transduction by RGD-fiber modified recombinant adenovirus into dendritic cells.
Dendritic cells (DC) are important antigen-presenting cells in the development of an anti-tumor T cell response. To extend the range of current immuno / gene therapies, we tested luciferase-expressing RGD-adenovirus (Ad) (Ad5lucRGD)-mediated transduction into DC. Phenotypically characterized DC were generated from peripheral blood CD14(+) cells by incubation with granulocyte-macrophage colony-stimulating factor, interleukin-4 and tumor necrosis factor alpha. On the 7th day of culture, the cells became mature DC with a CD1a(+), CD11c(+), CD80(+), CD83(+), CD86(+), human leukocyte antigen (HLA)-DR(+), CD14- phenotype. The expression of alpha( v)beta(3) integrin was enhanced on day 3 and returned to the basal level on day 7. We then compared the transduction efficiency of an Ad5lucRGD system to that using conventional Ad, in cells harvested on days 1, 3 and 7 of culture. Luciferase activity was negligible in AdCMVLuc, but remarkable in cells processed with Ad5lucRGD. Activity was maximal in cells that had been cultured for 3 days. Recombinant Ad5 fiber knob protein blocked AdCMVLuc- and Ad5lucRGD-mediated gene transduction by 90% and 20%, respectively. Surface markers and cytokine production were not affected by Ad5lucRGD-mediated transduction.
Asada-Mikami
R
R
Pharmacology Division, National Cancer Center Hospital, Chuo-ku, Tokyo 104-0045, Japan.
Heike
Y
Y
Kanai
S
S
Azuma
M
M
Shirakawa
K
K
Takaue
Y
Y
Krasnykh
V
V
Curiel
D T
DT
Terada
M
M
Abe
T
T
Wakasugi
H
H
D000256:Adenoviridae / Q000235:genetics*; D000595:Amino Acid Sequence; D015703:Antigens, CD / Q000032:analysis; D018929:Cell Culture Techniques / Q000379:methods; D002478:Cells, Cultured; D003713:Dendritic Cells / Q000166:cytology* / Q000187:drug effects / Q000276:immunology; D005434:Flow Cytometry; D017930:Genes, Reporter; D005822:Genetic Vectors; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D006801:Humans; D016130:Immunophenotyping; D016023:Integrins / Q000032:analysis; D015847:Interleukin-4 / Q000494:pharmacology; D018950:Lipopolysaccharide Receptors / Q000032:analysis; D008156:Luciferases / Q000032:analysis / Q000235:genetics*; D008175:Lung Neoplasms; D009842:Oligopeptides; D014162:Transfection / Q000379:methods; D014407:Tumor Cells, Cultured; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
D015703:Antigens, CD; D016023:Integrins; D018950:Lipopolysaccharide Receptors; D009842:Oligopeptides; D014409:Tumor Necrosis Factor-alpha; D015847:Interleukin-4; C047981:arginyl-glycyl-aspartic acid; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D008156:Luciferases