title: HHV-8/KSHV during the development of Kaposi's sarcoma: evaluation by polymerase chain reaction and immunohistochemistry.
abstract: The human gamma-herpes virus-8 (HHV-8) was first described in AIDS-related Kaposi's sarcoma (KS) tumour samples. In this study, we report comparative studies on paraffin-embedded biopsies of AIDS-related KS (AKS) and endemic KS (EKS) with regard to HHV-8 content as evaluated using polymerase chain reaction (PCR) and immunohistochemistry. DNA was extracted either using Chelex-100 or using Qia-gene kit and was evaluated with the help of a semiquantitative PCR assay. The PCR detection of HHV-8 was more sensitive to the Chelex method than to Qia-gene. The threshold for PCR test sensitivity with the help of serial dilution of DNA was at the level of five plasmid ORF-26 regions, and DNA from 25 body cavity-based lymphoma-1 cells. The results expressed as virus load/actin unit showed progressively higher HHV-8 levels in late (nodular) cases, compared to those in early (patch/plaque) stages. Evaluation of HHV-8 DNA levels in tumour tissues, thus, indicates a correlation between virus load and KS stage. Double immunostaining of spindle cells (SC) in KS biopsies for CD34 and HHV-8/latency-associated nuclear antigen (LANA) showed an increase in double-positive SC in the lesions of nodular AKS and EKS cases, compared to that in plaque and patch stages. However, 10-15% of CD34+/LANA- SC cells were observed during the development from patch to nodular cases of AKS and EKS. Our results indicate that PCR analysis is a simple and sensitive diagnostic method for HHV-8 evaluation in KS tissues, processed for conventional histopathology.
authors.affiliation: Immunopathology Laboratory, Cancer Center Karolinska, Karolinska Hospital, Solna, Stockholm, Sweden. parpak@ki.se
authors.forename: Fatemeh
authors.initials: F
authors.lastname: Pak
authors.forename: Pawan
authors.initials: P
authors.lastname: Pyakural
authors.forename: Parviz
authors.initials: P
authors.lastname: Kokhaei
authors.forename: Ephata
authors.initials: E
authors.lastname: Kaaya
authors.forename: A A
authors.initials: AA
authors.lastname: Pourfathollah
authors.forename: Galina
authors.initials: G
authors.lastname: Selivanova
authors.forename: Peter
authors.initials: P
authors.lastname: Biberfeld
mesh_terms: D000163:Acquired Immunodeficiency Syndrome / Q000150:complications; D018952:Antigens, CD34 / Q000032:analysis; D000956:Antigens, Viral / Q000032:analysis; D002452:Cell Count; D004273:DNA, Neoplasm / Q000032:analysis; D004279:DNA, Viral / Q000302:isolation & purification*; D019288:Herpesvirus 8, Human / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D006801:Humans; D007150:Immunohistochemistry; D009687:Nuclear Proteins / Q000032:analysis; D016133:Polymerase Chain Reaction; D012514:Sarcoma, Kaposi / Q000276:immunology / Q000473:pathology / Q000821:virology*; D012878:Skin Neoplasms / Q000276:immunology / Q000473:pathology / Q000821:virology*
chemical_list: D018952:Antigens, CD34; D000956:Antigens, Viral; D004273:DNA, Neoplasm; D004279:DNA, Viral; D009687:Nuclear Proteins; C109540:latency-associated nuclear antigen