Pak F., Pyakural P., Kokhaei P., Kaaya E., Pourfathollah A.A., Selivanova G., Biberfeld P.
Immunopathology Laboratory, Cancer Center Karolinska, Karolinska Hospital, Solna, Stockholm, Sweden; Immune and Gene Therapy Laboratory, Cancer Center Karolinska, Karolinska Hospital, Solna, Stockholm, Sweden; Department of Pathology, Muhimbili Univ. Coll. of Hlth. Sci., Dar-Es-Salaam, Tanzania; Department of Immunology, Tarbiat Modaress University, Blood Transfusion Organization, Tehran, Iran; Microbiol. and Tumor Biology Center, Karolinska Institutet, Stockholm, Sweden; Immunopathology Laboratory, Karolinska Hospital, 171-76 Solna, Stockholm, Sweden
Pak, F., Immunopathology Laboratory, Cancer Center Karolinska, Karolinska Hospital, Solna, Stockholm, Sweden, Immunopathology Laboratory, Karolinska Hospital, 171-76 Solna, Stockholm, Sweden; Pyakural, P., Immunopathology Laboratory, Cancer Center Karolinska, Karolinska Hospital, Solna, Stockholm, Sweden; Kokhaei, P., Immune and Gene Therapy Laboratory, Cancer Center Karolinska, Karolinska Hospital, Solna, Stockholm, Sweden; Kaaya, E., Department of Pathology, Muhimbili Univ. Coll. of Hlth. Sci., Dar-Es-Salaam, Tanzania; Pourfathollah, A.A., Department of Immunology, Tarbiat Modaress University, Blood Transfusion Organization, Tehran, Iran; Selivanova, G., Microbiol. and Tumor Biology Center, Karolinska Institutet, Stockholm, Sweden; Biberfeld, P., Immunopathology Laboratory, Cancer Center Karolinska, Karolinska Hospital, Solna, Stockholm, Sweden
The human γ-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.
article; cancer staging; clinical article; comparative study; controlled study; correlation analysis; DNA determination; DNA extraction; histopathology; human; Human herpesvirus 8; human tissue; immunofluorescence; immunohistochemistry; intermethod comparison; Kaposi sarcoma; polymerase chain reaction; sensitivity and specificity; spindle cell; tumor biopsy; virus detection; virus load; Acquired Immunodeficiency Syndrome; Antigens, CD34; Antigens, Viral; Cell Count; DNA, Neoplasm; DNA, Viral; Herpesvirus 8, Human; Humans; Immunohistochemistry; Nuclear Proteins; Polymerase Chain Reaction; Sarcoma, Kaposi; Skin Neoplasms