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

Hiroshi Kimura, Kanae Miyake, Yohei Yamauchi, Kana Nishiyama, Seiko Iwata, Keiji Iwatsuki, Kensei Gotoh, Seiji Kojima, Yoshinori Ito, Yukihiro Nishiyama

Research output: Contribution to journalArticle (Academic Journal)

41 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)1078-87
Number of pages10
JournalJournal of Infectious Diseases
Volume200
Issue number7
DOIs
Publication statusPublished - 1 Oct 2009

Keywords

  • Adolescent
  • Cell Line, Tumor
  • Child
  • Flow Cytometry
  • Herpesvirus 4, Human
  • Humans
  • Hydroa Vacciniforme
  • In Situ Hybridization, Fluorescence
  • Lymphocyte Subsets
  • Lymphoproliferative Disorders
  • Male
  • Sensitivity and Specificity
  • Transplantation
  • Journal Article
  • Research Support, Non-U.S. Gov't

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