Identification of Epstein-Barr Virus (EBV)-Infected Lymphocyte Subtypes by Flow Cytometric In Situ Hybridization in EBV-Associated Lymphoproliferative Diseases
Identification of Epstein-Barr Virus (EBV)-Infected Lymphocyte Subtypes by Flow Cytometric In Situ Hybridization in EBV-Associated Lymphoproliferative Diseases
复制标题
DOI：
10.1086/605610
复制
发表时间：
2009-10-01
影响因子：
6.4
通讯作者：
Nishiyama, Yukihiro
中科院分区：
文献类型：
--
作者：
Kimura, Hiroshi;Miyake, Kanae;Nishiyama, Yukihiro
为了诊断eb病毒（EBV）相关疾病并探讨EBV感染的发病机制，不仅需要测量EBV载量，还必须鉴定EBV感染的细胞。我们建立了一种新的流式细胞术原位杂交检测方法，使用EBV编码小RNA （EBER）特异性肽核酸探针检测EBV（+）悬浮细胞。通过增强荧光和光稳定性，我们成功地在同一细胞上染色了EBER和表面抗原。在3例疫苗形水疱样淋巴细胞增生性疾病患者中，我们证实1.7%-25.9%的外周血淋巴细胞感染EBV，并特异性鉴定这些淋巴细胞为CD3(+)CD4(-)CD8(-) γ δ T细胞受体阳性T细胞。结果表明，这种新颖的无创检测方法是一种直接、可靠的表征EBV感染淋巴细胞的方法，不仅可用于诊断EBV感染，还可用于阐明EBV相关疾病的发病机制。
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(-) gamma delta 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.