Quantitative analysis of latent human cytomegalovirus.
Cytomegalovirus latency depends on an interaction with hematopoietic cells in bone marrow and peripheral blood. The distribution of viral DNA was investigated by PCR-driven in situ hybridization (PCR-ISH), and the number of viral genomes per cell was estimated by quantitative competitive PCR during both experimental and natural latent infection. During experimental latent infection of cultured granulocyte-macrophage progenitors, the viral genome was detected in >90% of cells at a copy number of 1 to 8 viral genomes per cell. During natural infection, viral genomes were detected in 0.004 to 0.01% of mononuclear cells from granulocyte colony-stimulating factor-mobilized peripheral blood or bone marrow from seropositive donors, at a copy number of 2 to 13 genomes per infected cell. When evaluated by reverse transcription-PCR-ISH, only a small proportion of experimentally infected cells (approximately 2%) had detectable latent transcripts. This investigation identifies the small percentage of bone marrow-derived mononuclear cells that become latently infected during natural infection and suggests that latency may proceed in some cells that fail to encode currently identified latent transcripts.
Slobedman
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Department of Microbiology and Immunology, Stanford University School of Medicine, Stanford, California 94305-5124, USA.
Mocarski
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D002478:Cells, Cultured; D003587:Cytomegalovirus / Q000235:genetics* / Q000502:physiology; D004279:DNA, Viral / Q000032:analysis*; D016679:Genome, Viral; D006412:Hematopoietic Stem Cells / Q000821:virology; D006801:Humans; D017403:In Situ Hybridization; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000032:analysis; D017735:Virus Latency
D004279:DNA, Viral; D012333:RNA, Messenger