Detection of immediate early, early and late antigens of human cytomegalovirus by flow cytometry.
The development of selective inhibitors of human cytomegalovirus (CMV) should lead to a better understanding of the mode of replication of CMV. A flow cytometric method was developed to monitor the expression of CMV antigens in CMV-infected human embryonic lung fibroblasts. The procedure is based on an indirect immunofluorescence assay using monoclonal antibodies directed against CMV-specific (immediate early, early, late) antigens and goat anti-murine IgG labeled with fluorescein isothiocyanate. Flow cytometric analysis clearly distinguished between the uninfected and infected cell population. There was a time-dependent appearance of CMV-specific antigens and a close correlation between the multiplicity of infection and the ratio of infected to uninfected cells. The method allows an accurate determination of the percentage of CMV-infected cells in the whole cell population and of the time of appearance of the viral antigens.
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Rega Institute for Medical Research, Katholieke Universiteit Leuven, Belgium.
Snoeck
R
R
Neyts
J
J
de Clercq
E
E
D000911:Antibodies, Monoclonal; D000956:Antigens, Viral / Q000032:analysis*; D002460:Cell Line; D003587:Cytomegalovirus / Q000276:immunology* / Q000502:physiology; D005347:Fibroblasts; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D006801:Humans; D017874:Immediate-Early Proteins; D013997:Time Factors
D000911:Antibodies, Monoclonal; D000956:Antigens, Viral; D017874:Immediate-Early Proteins; C044582:immediate-early proteins, cytomegalovirus