title: Development of adenovirus vectors encoding rat complement regulators for use in therapy in rodent models of inflammatory diseases.
abstract: C activation has been implicated in the pathogenesis of numerous inflammatory human diseases and disease models. A therapy based on C inhibition might therefore be of benefit to reduce inflammation and ameliorate disease. C inhibition in vivo can be accomplished by the delivery of soluble recombinant C regulators either systemically or directly to a target site, but effects are transitory. We have developed a strategy for the efficient delivery of the membrane-bound rat C inhibitors, CD59, Crry, and decay-accelerating factor (DAF), using replication-deficient adenovirus vectors with the intention of treating rat models of disease in which C is implicated. The adenovirus recombinants(RAd), RAdCD59, RAdCrry, and RAdDAF, respectively, have been tested for expression and function of the transgene in vitro. Infection of human fetal foreskin fibroblasts resulted in high levels of expression of each of the rat inhibitors. The constructs were also tested for inhibition of rat C-mediated cell lysis and C3b deposition. In a cell lysis assay, each inhibited to varying degrees of efficiency in the order RAdCD59 = RAdDAF > RAdCrry. In a C3b deposition assay, RAdDAF caused a greater reduction in C3b deposition than RAdCrry and RAdCD59 was ineffective. These agents, individually or in combination, provide the tools for testing the effects of prolonged inhibition of C at a target site on the progress of experimental models of disease.
authors.affiliation: Department of Medical Biochemistry, University of Wales College of Medicine, Cardiff, United Kingdom.
authors.forename: Y
authors.initials: Y
authors.lastname: McGrath
authors.forename: G W
authors.initials: GW
authors.lastname: Wilkinson
authors.forename: O B
authors.initials: OB
authors.lastname: Spiller
authors.forename: B P
authors.initials: BP
authors.lastname: Morgan
mesh_terms: D000256:Adenoviridae / Q000235:genetics* / Q000276:immunology*; D000818:Animals; D000954:Antigens, Surface; D018958:CD55 Antigens / Q000096:biosynthesis / Q000235:genetics; D018957:CD59 Antigens / Q000096:biosynthesis / Q000235:genetics; D002461:Cell Line, Transformed; D003176:Complement C3 / Q000037:antagonists & inhibitors / Q000378:metabolism; D003169:Complement Inactivator Proteins / Q000096:biosynthesis / Q000235:genetics*; D003602:Cytotoxicity, Immunologic; D004195:Disease Models, Animal; D004306:Dose-Response Relationship, Immunologic; D005347:Fibroblasts / Q000276:immunology / Q000378:metabolism / Q000821:virology; D005822:Genetic Vectors / Q000008:administration & dosage / Q000138:chemical synthesis / Q000276:immunology / Q000627:therapeutic use*; D017261:Glycosylphosphatidylinositols / Q000378:metabolism; D006801:Humans; D007249:Inflammation / Q000235:genetics* / Q000276:immunology / Q000473:pathology / Q000628:therapy*; D007269:Injections, Intra-Arterial; D043265:Phosphatidylinositol Diacylglycerol-Lyase; D051381:Rats; D011956:Receptors, Cell Surface; D011951:Receptors, Complement / Q000096:biosynthesis / Q000235:genetics; D013194:Staining and Labeling; D013583:Synovial Membrane / Q000201:enzymology / Q000473:pathology; D010738:Type C Phospholipases / Q000494:pharmacology; D001616:beta-Galactosidase / Q000008:administration & dosage / Q000032:analysis
chemical_list: D000954:Antigens, Surface; D018958:CD55 Antigens; D018957:CD59 Antigens; D003176:Complement C3; D003169:Complement Inactivator Proteins; C513105:Cr1l protein, rat; D017261:Glycosylphosphatidylinositols; D011956:Receptors, Cell Surface; D011951:Receptors, Complement; D010738:Type C Phospholipases; D001616:beta-Galactosidase; D043265:Phosphatidylinositol Diacylglycerol-Lyase