title: Expression of foreign genes in cultured human primary macrophages using recombinant vaccinia virus vectors.
abstract: Recombinant vaccinia viruses (re-VVs) provide an extremely versatile method for the expression of foreign genes in a wide range of cultured cell types of different lineages and species. In the present report, we examine the utility of re-VV vectors for re-protein production in cultured human primary macrophages obtained through in vitro differentiation of peripheral blood monocytes. Primary macrophages supported early stages of the VV infection cycle, including morphologic cytopathic effect, shut-off of host protein synthesis and activation of early viral protein synthesis; however, late stages of infection were blocked, including synthesis of late viral proteins, replication of viral DNA, and production of infectious progeny virions. Abortive infection was observed with several independent VV strains. Using re-VVs containing Escherichia coli lacZ as a reporter gene, we assayed the activities of different classes of VV promoters. Consistent with the results noted above, human primary macrophages supported reporter gene expression driven by an early or intermediate VV promoter, but not by a late promoter; expression was obtained with synthetic bifunctional promoters containing early and/or intermediate components. Primary macrophages also supported the VV/bacteriophage T7 RNA polymerase hybrid gene expression system. The utility of re-VV vectors for production of proteins of biological interest in human primary macrophages was demonstrated using re-VVs encoding human CD4 and the human immunodeficiency virus type-1 envelope glycoprotein.
authors.affiliation: Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892.
authors.forename: C C
authors.initials: CC
authors.lastname: Broder
authors.forename: P E
authors.initials: PE
authors.lastname: Kennedy
authors.forename: F
authors.initials: F
authors.lastname: Michaels
authors.forename: E A
authors.initials: EA
authors.lastname: Berger
mesh_terms: D015704:CD4 Antigens / Q000096:biosynthesis / Q000235:genetics; D002478:Cells, Cultured; D004279:DNA, Viral / Q000032:analysis; D012321:DNA-Directed RNA Polymerases / Q000235:genetics; D015967:Gene Expression Regulation, Viral; D015686:Gene Products, env / Q000096:biosynthesis / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics*; D015488:HIV Antigens / Q000096:biosynthesis / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D008264:Macrophages / Q000382:microbiology*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics; D014616:Vaccinia virus / Q000235:genetics* / Q000254:growth & development; D014764:Viral Proteins
chemical_list: D015704:CD4 Antigens; D004279:DNA, Viral; D015686:Gene Products, env; D015488:HIV Antigens; D011994:Recombinant Proteins; D014764:Viral Proteins; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases