title: Visualization of human herpesvirus type 8 in Kaposi's sarcoma by light and transmission electron microscopy.
abstract: 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.
  
  
  METHODS
  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 (Tl.1 riboprobe) by in situ hybridization (ISH), for viral inclusions by light microscopy, and for herpesviruses by transmission electron microscopy (TEM). Sections were also labeled with Tl.1 counterstained with CD34, an endothelial cell marker.
  
  
  RESULTS
  The skin lesion was DNA PCR-positive for HHV-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 revealed infection by a virus displaying the typical morphology and cytopathicity of herpesviruses. Hexagonal nucleocapsids and mature enveloped virions were present in vasoformative spindle cells and mononuclear cells, often resembling lymphocytes. Extrapolating from TEM to standard light microscopy on hematoxylin and eosin-stained paraffin sections, eosinophilic, targetoid intranuclear inclusions were identified within spindle cells which often lined vascular lumina. The Tl.1-riboprobe labeled CD34+ spindle cells containing intranuclear inclusions, as well as mononuclear cells within Kaposi's sarcoma and residual lymphoid tissue.
  
  
  CONCLUSIONS
  The herpesvirus visualized in Kaposi's sarcoma lesions has morphologic and cytopathic features typical of human herpesviruses, productively infects vasoformative spindle cells and mononuclear cells, and is consistent with HHV-8. It can also form intranuclear inclusions that are identifiable by light microscopy in hematoxylin and eosin sections and by ISH.
authors.affiliation: Department of Pathology, George Washington University, Washington, DC, USA.
authors.forename: J M
authors.initials: JM
authors.lastname: Orenstein
authors.forename: S
authors.initials: S
authors.lastname: Alkan
authors.forename: A
authors.initials: A
authors.lastname: Blauvelt
authors.forename: K T
authors.initials: KT
authors.lastname: Jeang
authors.forename: M D
authors.initials: MD
authors.lastname: Weinstein
authors.forename: D
authors.initials: D
authors.lastname: Ganem
authors.forename: B
authors.initials: B
authors.lastname: Herndier
mesh_terms: D019288:Herpesvirus 8, Human / Q000302:isolation & purification / Q000648:ultrastructure*; D006801:Humans; D017403:In Situ Hybridization; D008297:Male; D008854:Microscopy, Electron; D012514:Sarcoma, Kaposi / Q000473:pathology / Q000648:ultrastructure / Q000821:virology*