VISUALIZATION OF HUMAN HERPESVIRUS TYPE-8 IN KAPOSIS-SARCOMA BY LIGHTAND TRANSMISSION ELECTRON-MICROSCOPY

Background: Human herpesvirus type 8 (HHV-8) has been associated with Kaposi's sarcoma, body cavity-based lymphoma (BCBL), and multicentric Castleman's disease through DNA, in situ hybridization, and serologic studies. HHV-8 has been visualized only in HHV-8-positive/Epstein-Barr virus (EBV)-negative/ cytomegalovirus (CMV)-negative BCBL cell lines, but not in HHV-8-positive/EBV-negative/CMV-negative Kaposi's sarcoma lesions. Design: Kaposi's sarcoma of the skin, lymph node, and spleen from three patients with AIDS were analysed for HHV-8, EBV and CMV DNA by polymerase chain reaction (PCR), for HHV-8 RNA (T1.1 riboprobe) by in situ hybridization (ISH), for viral inclusions by light microscopy , and for herpesviruses by transmission electron microscopy (TEM). Sec tions were also labeled with T1.1 counterstained with CD34, an endothe lial cell marker. Results: The skin lesion was DNA PCR-positive for HH V-8 and CMV (nested, but not single PCR), the lymph node was positive for HHV-8 and EBV, and the spleen was positive for only HHV-8. TEM rev ealed infection by a virus displaying the typical morphology and cytop athicity of herpesviruses. Hexagonal nucleocapsids and mature envelope d virions were present in vasoformative spindle cells and mononuclear cells, often resembling lymphocytes. Extrapolating from TEM to standar d light microscopy on hematoxylin and eosin-stained paraffin sections, eosinophilic, targetoid intranuclear inclusions were identified withi n spindle cells which often lined vascular lumina. The T1.1-riboprobe labeled CD34+ spindle cells containing intranuclear inclusions, as wel l as mononuclear cells within Kaposi's sarcoma and residual lymphoid t issue. Conclusion: The herpesvirus visualized in Kaposi's sarcoma lesi ons has morphologic and cytopathic features typical of human herpesvir uses, productively infects vasoformative spindle cells and mononuclear cells, and is consistent with HHV-8. It can also form intranuclear in clusions that are identifiable by light microscopy in hematoxylin and eosin sections and by ISH.