Expression of foreign genes in cultured human primary macrophages using recombinant vaccinia virus vectors.

Important note: Information in this article was accurate in 1994. The state of the art may have changed since the publication date.

Expression of foreign genes in cultured human primary macrophages using recombinant vaccinia virus vectors.

Gene. 1994 May 16;142(2):167-74. Unique Identifier : AIDSLINE MED/94252563
Broder CC; Kennedy PE; Michaels F; Berger EA; Laboratory of Viral Diseases, National Institute of Allergy and; Infectious Diseases, National Institutes of Health, Bethesda, MD; 20892.

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.

Keywords: Antigens, CD4/BIOSYNTHESIS/GENETICS Cells, Cultured DNA, Viral/ANALYSIS Gene Expression Regulation, Viral Gene Products, env/BIOSYNTHESIS/GENETICS Genetic Vectors/*GENETICS Hela Cells Human HIV Antigens/BIOSYNTHESIS/GENETICS Macrophages/*MICROBIOLOGY Promoter Regions (Genetics)/*GENETICS Recombinant Proteins/*BIOSYNTHESIS/GENETICS RNA Polymerases/GENETICS Support, Non-U.S. Gov't Support, U.S. Gov't, P.H.S. Vaccinia Virus/GROWTH & DEVELOPMENT/*GENETICS JOURNAL ARTICLE

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