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Blood, 1 August 2002, Vol. 100, No. 3, pp. 888-896
HEMOSTASIS, THROMBOSIS, AND VASCULAR BIOLOGY
Characterization of Kaposi sarcoma-associated herpesvirus/human
herpesvirus-8 infection of human vascular endothelial cells:
early events
Bruce J. Dezube,
Maria Zambela,
David R. Sage,
Jian-Feng Wang, and
Joyce D. Fingeroth
From the Divisions of Infectious Disease, Experimental
Medicine, and Hematology/Oncology, Department of Medicine, Beth Israel
Deaconess Medical Center and Harvard Medical School, Boston, MA.
Kaposi sarcoma-associated herpesvirus (KSHV)/human herpesvirus-8
(HHV-8) is causally associated with Kaposi sarcoma (KS). The absence of
a cell culture system that effectively reproduces the composite
mechanisms governing initiation and maintenance of HHV-8 infection
(lytic and latent) in KS endothelial cells, however, has left important
questions unanswered. Here, we report a culture system in which the
earliest events that accompany HHV-8 infection could be surveyed in
primary endothelial cells. Binding of HHV-8 to microvascular dermal
endothelial cells (MVDECs) was directly compared with other primary
target cells implicated in HHV-8-associated diseases. Virus
attachment, fusion, internalization and transport within MVDECs was
monitored by electron microscopy. Studies of genome configuration
revealed that rapid circularization of the viral DNA occurred on entry,
though by 72 hours after infection linear DNAs accumulated and early as
well as late lytic RNAs (T1.1, K8.1) could be detected. The latency
transcripts (LT1/LT2) were first detected on day 8, demonstrating that
both lytic and latent infection were initiated. Although most lytic
transcripts accrued until passage, open-reading frame-74 RNAs
fluctuated with a fixed periodicity, suggesting that early replication
after infection of MVDECs was synchronous.
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