       Document 0121
 DOCN  M9470121
 TI    Expression of foreign genes in cultured human primary macrophages using
       recombinant vaccinia virus vectors.
 DT    9409
 AU    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.
 SO    Gene. 1994 May 16;142(2):167-74. Unique Identifier : AIDSLINE
       MED/94252563
 AB    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.
 DE    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

       SOURCE: National Library of Medicine.  NOTICE: This material may be
       protected by Copyright Law (Title 17, U.S.Code).

