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Demonstration of Kaposi's Sarcoma-Associated Herpes Virus Cyclin D
Homolog in Cutaneous Kaposi's Sarcoma by Colorimetric In Situ
Hybridization Using a Catalyzed Signal Amplification System
Jon A. Reed,
Roland G. Nador,
David Spaulding,
Yoichi Tani,
Ethel Cesarman, and
Daniel M. Knowles
From the Department of Pathology, The New York Hospital-Cornell
Medical Center, New York, NY; and Dako Corp, Carpinteria, CA.
Kaposi's sarcoma-associated herpes virus (KSHV)/human herpes virus
8 (HHV8) DNA sequences have been demonstrated in Kaposi's sarcoma
(KS), as well as in some acquired immunodeficiency syndrome (AIDS)-related non-Hodgkin's lymphomas (NHL) and in multicentric Castleman's disease. Although KSHV DNA generally is abundant in KSHV-associated lymphomas, few copies of the virus are present in KS, a
property that confounds detection by in situ methods. Previous in situ
studies, which identified KSHV in lesions of KS, relied on the use of
polymerase chain reaction (PCR) to amplify target DNA sequences before
in situ hybridization (ISH) for localization or used ISH with
radioactively-labeled probes to obtain adequate levels of detection
sensitivity. In this study, a novel nonisotopic nucleic acid ISH method
using catalyzed signal amplification and colorimetric detection without
PCR-dependent target amplification was used to identify KSHV-specific
sequences. The level of sensitivity was increased further by using a
probe that detects viral cyclin D homolog transcripts, which are
expressed at significant levels during latent viral infection. Thirty
cutaneous lesions of KS (25 AIDS-related and five classical European
type) were evaluated. AIDS-related NHL and cell lines derived from
patients with AIDS-related NHL, all of which were known to harbor KSHV
by Southern blot analysis, were used as positive controls. NHL and
benign cutaneous vascular lesions not associated with AIDS were used as
negative controls. For each of the 30 KS lesions studied, hybridization
signals were detected in most of the spindle cells surrounding the
atypical slit-like vascular channels and also were detected in some
endothelial cells in well-formed blood vessels in the perilesional
dermis. Plaque and nodular lesions generally contained more labeled
cells than did early patch lesions. All AIDS-related NHL and cell lines contained KSHV-specific sequences; however, the non-AIDS-related NHLs
and benign vascular lesions were negative. These results confirm the
presence of KSHV sequences in cutaneous KS and provide in situ evidence
of infection by this virus in early patch-stage lesions. This study
also defines the in situ expression of the KSHV cyclin D homolog viral
oncogene in cutaneous KS. The use of this sensitive nonisotopic ISH
method should allow detection of other KSHV-specific gene products,
further defining the pathobiology of this virus.
Blood, Vol. 91 No. 10 (May 15), 1998:
pp. 3825-3832
© 1998 by The American Society of Hematology.
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