title: Incorporation of adenovirus into a ligand-based DNA carrier system results in retention of original receptor specificity and enhances targeted gene expression.
abstract: Adenovirus type 5 was modified by coupling an asialoglycoprotein-polylysine conjugate to the virus by reactions that activate carbohydrate residues. Wild-type virus modified in this manner had greatly decreased infectivity toward normally susceptible HeLa S3 (asialoglycoprotein receptor (-)) and SK Hep1 (asialoglycoprotein receptor (-)) cells leaving 91 and 86% viable, respectively, after 48 h. However, with Huh 7 (asialoglycoprotein receptor (+)) cells, modified virus retained its infectivity leaving only 19% of cells viable under identical conditions. Modified virus was complexed to DNA in the form of a plasmid, pSVHBV surf, containing the gene for hepatitis B surface antigen as a marker of gene expression. Huh 7, receptor (+), cells treated with modified wild type, and modified replication-defective d1312 virus complexed to DNA raised antigen levels by approximately 13- and 30-fold, respectively, compared with asialoglycoprotein-polylysine DNA complex alone. Competition with a large excess of an asialoglycoprotein blocked the enhancement by more than 95%. Using a beta-galactosidase marker gene, the number of cells transfected by modified virus was found to be 200-fold higher than complex alone. Yet, specificity was retained exclusively for asialoglycoprotein receptor-bearing cells. These data indicate that adenovirus can be chemically modified by coupling ligands resulting in targeted gene expression dictated specifically by receptor recognition of the attached ligand.
authors.affiliation: Department of Medicine, University of Connecticut School of Medicine, Farmington 06030.
authors.forename: G Y
authors.initials: GY
authors.lastname: Wu
authors.forename: P
authors.initials: P
authors.lastname: Zhan
authors.forename: L L
authors.initials: LL
authors.lastname: Sze
authors.forename: A R
authors.initials: AR
authors.lastname: Rosenberg
authors.forename: C H
authors.initials: CH
authors.lastname: Wu
mesh_terms: D000260:Adenoviruses, Human / Q000235:genetics*; D037263:Asialoglycoprotein Receptor; D001212:Asialoglycoproteins; D002467:Cell Nucleus / Q000378:metabolism; D004337:Drug Carriers; D005819:Genetic Markers; D005822:Genetic Vectors; D006367:HeLa Cells; D006514:Hepatitis B Surface Antigens / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D010957:Plasmids; D011107:Polylysine / Q000031:analogs & derivatives; D011956:Receptors, Cell Surface / Q000378:metabolism*; D014162:Transfection / Q000379:methods*; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000235:genetics
chemical_list: D037263:Asialoglycoprotein Receptor; D001212:Asialoglycoproteins; D004337:Drug Carriers; D005819:Genetic Markers; D006514:Hepatitis B Surface Antigens; D011956:Receptors, Cell Surface; C086457:asialoglycoprotein-polylysine conjugate; D011107:Polylysine; D001616:beta-Galactosidase