Identification of Epstein-Barr virus (EBV)-infected lymphocyte subtypes by flow cytometric in situ hybridization in EBV-associated lymphoproliferative diseases.
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.
Kimura
Hiroshi
H
Department of Virology, Nagoya University Graduate School of Medicine, Nagoya, Japan. hkimura@med.nagoya-u.ac.jp
Miyake
Kanae
K
Yamauchi
Yohei
Y
Nishiyama
Kana
K
Iwata
Seiko
S
Iwatsuki
Keiji
K
Gotoh
Kensei
K
Kojima
Seiji
S
Ito
Yoshinori
Y
Nishiyama
Yukihiro
Y
D000293:Adolescent; D045744:Cell Line, Tumor; D002648:Child; D005434:Flow Cytometry; D004854:Herpesvirus 4, Human / Q000502:physiology*; D006801:Humans; D006837:Hydroa Vacciniforme / Q000821:virology; D017404:In Situ Hybridization, Fluorescence; D016131:Lymphocyte Subsets / Q000166:cytology* / Q000821:virology*; D008232:Lymphoproliferative Disorders / Q000209:etiology / Q000821:virology; D008297:Male; D012680:Sensitivity and Specificity; D014180:Transplantation / Q000009:adverse effects