Use of a GFP-PML-expressing cell line as a biosensor for human cytomegalovirus infection.
Human cytomegalovirus (HCMV) infection has a marked effect on promyelocytic leukemia (PML) bodies. Here, we describe a novel real-time monitoring system for HCMV-infected cells in vitro using a newly established cell line that stably expresses GFP-PML protein. Upon infection, HCMV causes specific dispersion of GFP-PML bodies, thereby allowing the infected cells to be monitored by fluorescence microscopy without immunostaining. Quantitative protocols using either an NPB fluorescence assay or a GFP-PML imaging assay are also described. The NPB fluorescence assay is rapid, sensitive, and sufficiently simple for screening of inhibitory reagents, while the GFP-PML imaging assay is highly sensitive and applicable to drug susceptibility testing of low-titer clinical isolates.
Ueno
Tomonori
T
Nippi Research Institute of Biomatrix, Toride, Ibaraki, Japan.
Ogawa-Goto
Kiyoko
K
D000818:Animals; D015374:Biosensing Techniques / Q000379:methods*; D002460:Cell Line; D002478:Cells, Cultured; D018920:Coculture Techniques; D006224:Cricetinae; D003586:Cytomegalovirus Infections / Q000235:genetics / Q000378:metabolism* / Q000821:virology*; D004198:Disease Susceptibility / Q000378:metabolism; D015870:Gene Expression / Q000235:genetics; D017930:Genes, Reporter / Q000235:genetics*; D006801:Humans; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D025521:Tumor Suppressor Proteins / Q000235:genetics / Q000378:metabolism*
D009687:Nuclear Proteins; D014157:Transcription Factors; D025521:Tumor Suppressor Proteins