Cytofluorographic analysis of effects of interferons on expression of human cytomegalovirus proteins.
The appearance of cytomegalovirus (CMV) proteins in infected fibroblasts was determined by flow cytometry. The sequential production of immediate early (IE), early (E), and late (L) proteins reacting with respective monoclonal antibodies (mAbs) E13, 58/5, and 24/4 was determined in fibroblasts infected with the AD-169 strain of CMV. The percentage of cells expressing CMV proteins and the intensity of fluorescence within cells were determined from day 1 to day 7 post-infection. The effect of interferons (IFNs) alpha, beta, gamma on expression of CMV proteins was analyzed using flow cytometry. IFNs inhibited E and L protein production at days 3 and 6 post-infection in a dose-dependent manner. This inhibitory effect on protein expression was associated with a reduction in release of infectious CMV into culture media. The method described here for detection of CMV proteins using flow cytometry may be useful for basic studies of gene expression and for diagnostic purposes.
Torigoe
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Division of Infectious Diseases and Immunology, Joseph Stokes, Jr. Research Institute of the Children's Hospital of Philadelphia, PA 19104.
Campbell
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Torigoe
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Michelson
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Starr
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D000911:Antibodies, Monoclonal; D002460:Cell Line; D003587:Cytomegalovirus / Q000187:drug effects* / Q000276:immunology / Q000502:physiology; D005069:Evaluation Studies as Topic; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D007370:Interferon Type I / Q000494:pharmacology; D016899:Interferon-beta / Q000494:pharmacology; D007371:Interferon-gamma / Q000494:pharmacology; D007372:Interferons / Q000494:pharmacology*; D011994:Recombinant Proteins; D014764:Viral Proteins / Q000032:analysis / Q000096:biosynthesis* / Q000276:immunology; D014773:Virology / Q000379:methods; D014779:Virus Replication / Q000187:drug effects
D000911:Antibodies, Monoclonal; D007370:Interferon Type I; D011994:Recombinant Proteins; D014764:Viral Proteins; D016899:Interferon-beta; D007371:Interferon-gamma; D007372:Interferons