Identification of Epstein-Barr virus (EBV)-infected lymphocyte subtypes by flow cytometric in situ hybridization in EBV-associated lymphoproliferative diseases.

Abstract

To diagnose Epstein-Barr virus (EBV)-associated diseases and to explore the pathogenesis of EBV infection, not only must the EBV load be measured, but EBV-infected cells must also be identified. We established a novel flow cytometric in situ hybridization assay to detect EBV(+) suspension cells using a peptide nucleic acid probe specific for EBV-encoded small RNA (EBER). By enhancing fluorescence and photostability, we successfully stained EBER and surface antigens on the same cells. In 3 patients with hydroa vacciniforme-like lymphoproliferative disease, we demonstrated that 1.7%-25.9% of peripheral lymphocytes were infected with EBV and specifically identified these lymphocytes as CD3(+)CD4(-)CD8(-) gammadelta T cell receptor-positive T cells. The results indicate that this novel and noninvasive assay is a direct and reliable method of characterizing EBV-infected lymphocytes that can be used not only to diagnose EBV infection but also to clarify the pathogenesis of EBV-associated diseases.

• Authors: Hiroshi Kimura, Kanae Miyake, Yohei Yamauchi, Kana Nishiyama, Seiko Iwata, Keiji Iwatsuki, Kensei Gotoh, Seiji Kojima, Yoshinori Ito, Yukihiro Nishiyama
• Journal: The Journal of infectious diseases (J Infect Dis) Vol. 200 Issue 7 Pg. 1078-87 (Oct 01 2009) ISSN: 0022-1899 [Print] United States
• PMID: 19698076 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Topics

• Adolescent
• Cell Line, Tumor
• Child
• Flow Cytometry
• Herpesvirus 4, Human (physiology)
• Humans
• Hydroa Vacciniforme (virology)
• In Situ Hybridization, Fluorescence
• Lymphocyte Subsets (cytology, virology)
• Lymphoproliferative Disorders (etiology, virology)
• Male
• Sensitivity and Specificity
• Transplantation (adverse effects)