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M9470121.TXT
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1994-07-02
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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).