title: Kinetic and biologic properties of recombinant ChIFN-gamma expressed via CELO-virus vector.
abstract: In this study, a replicative fowl adenovirus serotype 1 (CELO) recombinant expressing chicken interferon-gamma (ChIFN-gamma) was constructed. In the engineered recombinant, the ChIFN-gamma gene was placed under the control of cytomegalovirus (CMV) promoter. The ChIFN-gamma expression cassette was inserted in the right end of the CELO genome (D fragment), which was able to carry the largest insertion of foreign DNA without affecting the replication functions of the vector. The recombinant ChIFN-gamma (rChIFN-gamma) produced in the CELO-virus expression system was characterized by comparing its biologic activities with that of rChIFN-gamma produced via the baculovirus expression system (Bac-ChIFN-gamma). CELO-ChIFN-gamma inhibited the replication of cytolytic virus in chicken embryo fibroblasts (CEFs) and activated macrophages in a better manner than did Bac-ChIFN-gamma . Moreover, the in vitro and in vivo stability of the CELO-derived rChIFN-gamma was considerably higher than that of the Bac-ChIFN-gamma. The CELO-ChIFN-gamma recombinant vector was able to replicate in vitro in the loghorn male hepatoma (LMH) hepatocyte cell line and to produce detectable levels of recombinant cytokine in supernatant as early as 90 min post-infection. Therefore, the CELO-virus expression system is an appropriate system for high-level expression of biologically active and stable ChIFN-gamma.
authors.affiliation: Avian Virology and Immunology Unit, Veterinary and Agrochemical Research Centre (VAR), 1180 Ukkel, Brussels, Belgium. farau@var.fgov.be
authors.forename: Fabienne
authors.initials: F
authors.lastname: Rauw
authors.forename: Bénédicte
authors.initials: B
authors.lastname: Lambrecht
authors.forename: Achille
authors.initials: A
authors.lastname: François
authors.forename: Patrick
authors.initials: P
authors.lastname: Langlois
authors.forename: Thierry
authors.initials: T
authors.lastname: van den Berg
mesh_terms: D000818:Animals; D000998:Antiviral Agents / Q000378:metabolism / Q000494:pharmacology*; D045744:Cell Line, Tumor; D002642:Chick Embryo; D025681:Fowl adenovirus A; D015870:Gene Expression; D007371:Interferon-gamma / Q000096:biosynthesis / Q000494:pharmacology*; D007700:Kinetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000494:pharmacology*
chemical_list: D000998:Antiviral Agents; D011994:Recombinant Proteins; D007371:Interferon-gamma