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Journal of Virology, June 1999, p. 4806-4812, Vol. 73, No. 6
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Quantitative Analysis of Latent Human
Cytomegalovirus
Barry
Slobedman and
Edward S.
Mocarski*
Department of Microbiology and Immunology,
Stanford University School of Medicine, Stanford, California
94305-5124
Received 17 December 1998/Accepted 2 March 1999
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.
*
Corresponding author. Mailing address: Department of
Microbiology and Immunology, Stanford University School of Medicine, Stanford, CA 94305-5124. Phone: (650) 723-6435. Fax: (650) 723-1606. E-mail: mocarski{at}stanford.edu.
Journal of Virology, June 1999, p. 4806-4812, Vol. 73, No. 6
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
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