Reduced toxicity, attenuated immunogenicity and efficient mediation of human p53 gene expression in vivo by an adenovirus vector with deleted E1-E3 and inactivated E4 by GAL4-TATA promoter replacement

Abstract

A recombinant adenovirus with deleted E1 and E3, and E4-mouse livers from days 3 to 28. Ten weeks after injection, inactivated by replacing the E4 promoter with a synthetic p53 gene expression was still detected in G4-treated promoter composed of a minimal TATA box and five con-C57BL/6 mice at similar levels, but was not detectable in sensus yeast GAL4-binding site elements was developed WT-treated mice. Vector-induced liver toxicity was evaluand used to express the human tumor suppresser gene ated by analyzing serum transaminases (SGOT and p53. The toxicity and immunogenicity of this vector and SGPT) activities. In all cases, SGOT and SGPT activities vector-mediated p53 gene expression in vivo were studied were markedly decreased in EG-treated C3H and C57BL/6 in immunocompetent C3H and C57BL/6 mice. Expression mice compared with those in EW-treated mice on days 3, of the late viral gene product, hexon protein, was observed 7 and 14 after injection. In C57BL/6 mice, the total antiin C3H and C57BL/6 mice injected with E4 wild-type adenoviral CTL activities were two-to three-fold higher in adenovirus constructs Adv-cmv-␤-Gal (BG), Adv-cmv-hp53 animals treated with EW vector than in those treated with (WT), and empty E1 − vector Adv-E4 (EW) 3 to 28 days EG vector. These results suggest that inactivation of the after injection, but was undetectable in mice treated with E4 promoter efficiently diminished the viral replication and E4 modified empty E1 − vector Adv-GAL4 (EG) or Adv-cmvthe late viral gene expression, reduced host immune hp53-GAL4 (G4). Expression of the p53 gene was response and consequently reduced toxicity and prolonged observed in both WT-and G4-injected C3H and C57BL/6 the duration of transgene expression in vivo.

A rapid and efficient method for purification of recombinant adenovirus with arginine–glycine–aspartic acid-modified fibers

“…Histopathologic examination of liver sections of mice systemically treated with iodixanol-purified Ad/RGDLacZ at doses between 1 × 10 11 to 1 × 10 12 VP showed changes that are commonly observed in animals treated with E-deleted adenovectors, including mild portal triaditis, mild hepatocellular degeneration (cytomegaly and karyomegaly), and apoptosis in 1-3% of hepatocytes. Similar histopathologic changes have been observed in animals treated with CsClpurified adenovectors at doses of less than 1 × 10 11 VP [24,26], suggesting that adenovectors purified by iodixanol are at least as safe as those purified by the CsCl method. …”

“…The expansion, purification, titration, and quality analyses of the adenoviruses were performed and analyzed at the Institutional Vector Core Facility at the University of Texas M. D. Anderson Cancer Center as described previously [23,24].…”

Chimpanzee-origin adenovirus vectors as vaccine carriers

“…Furthermore, deletion of E4 minimizes outgrowth of replication-competent virus in packaging cell lines, although this is not a major concern for simianorigin Ad vectors grown on packaging cell lines containing the E1 of AdHu5 virus. Previous gene therapy trials reported that deletion or functional inactivation of E4 in AdHu5 vectors reduces the inflammatory response to the vector, 34 and decreases AdHu5-mediated upregulation of adhesion molecules in endothelial cells. 35 Such adhesion molecules are crucial for lymphocyte trafficking.…”

Adenoviral gene transfer of endothelial nitric oxide synthase (eNOS) to the penis improves age-related erectile dysfunction in the rat

“…25,26 However, they have also been shown to cause host in¯ammatory and immune responses that shorten the duration of gene expression in vivo. 27 Any successful gene therapy approach to human disease requires a therapeutic gene. NO plays a primary role in the process of penile erection.…”