Factors involved in the maturation of murine dendritic cells transduced with adenoviral vector variants.
Adenoviral vector (Ad)-mediated gene transfer is an attractive method for manipulating the immunostimulatory properties of dendritic cells (DCs) for cancer immunotherapy. DCs treated with Ad have phenotype alterations (maturation) that facilitate T cell sensitization. We investigated the mechanisms of DC maturation with Ad transduction. Expression levels of a maturation marker (CD40) on DCs treated with conventional Ad, fiber-modified Ads (AdRGD, AdF35, AdF35DeltaRGD), or a different serotype Ad (Ad35) were correlated with their transduction efficacy. The alphav-integrin directional Ad, AdRGD, exhibited the most potent ability to enhance both foreign gene expression and CD40 expression, and induced secretion of interleukin-12, tumor necrosis factor-alpha, and interferon-alpha in DCs. The presence of a foreign gene expression cassette in AdRGD was not necessary for DC maturation. Maturation of DCs treated with AdRGD was suppressed by destruction of the Ad genome, inhibition of endocytosis, or endosome acidification, whereas proteasome inhibition increased CD40 expression levels on DCs. Moreover, inhibition of alphav-integrin signal transduction and blockade of cytokine secretion affected the maturation of DCs treated with AdRGD only slightly or not at all, respectively. Thus, our data provide evidence that Ad-induced DC maturation is due to Ad invasion of the DCs, followed by nuclear transport of the Ad genome, and not to the expression of foreign genes.
Kanagawa
Naoko
N
Department of Biotechnology and Therapeutics, Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka 565-0871, Japan.
Koretomo
Ryosuke
R
Murakami
Sayaka
S
Sakurai
Fuminori
F
Mizuguchi
Hiroyuki
H
Nakagawa
Shinsaku
S
Fujita
Takuya
T
Yamamoto
Akira
A
Okada
Naoki
N
D000256:Adenoviridae / Q000145:classification / Q000235:genetics / Q000472:pathogenicity; D000818:Animals; D019013:CD40 Antigens / Q000235:genetics / Q000378:metabolism; D002454:Cell Differentiation; D003713:Dendritic Cells / Q000166:cytology* / Q000821:virology*; D005260:Female; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D014161:Transduction, Genetic; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism
D019013:CD40 Antigens; D014764:Viral Proteins; D049452:Green Fluorescent Proteins