| Project/Area Number |
10670709
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Pediatrics
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| Research Institution | Kanazawa University |
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Principal Investigator |
OHTA Kazuhide Kanazawa University University Hospital, Department of Pediatrics, Assistant Professor, 医学部・附属病院, 講師 (20283129)
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| Co-Investigator(Kenkyū-buntansha) |
YACHIE Akihiro Kanazawa University Faculty of Medicine, Department of Laboratory, Professor, 医学部, 教授 (40210281)
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| Project Period (FY) |
1998 – 1999
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| Project Status |
Completed (Fiscal Year 1999)
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| Budget Amount *help |
¥3,100,000 (Direct Cost: ¥3,100,000)
Fiscal Year 1999: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1998: ¥2,000,000 (Direct Cost: ¥2,000,000)
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| Keywords | EB virus (EBV) / PCR method / EBV terminal repeat / EBER-1 / in situ hybridization / EBウイルス(EBV) / PCR法 / EVE terminal repeat / in situ hubridization |
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| Research Abstract |
Epstein-Barr virus (EBV) is prevalent among adult population and it persists within B lymphocytes in latent forms throughout life. Infection. mononucleosis is a well recognized, self-limited EBV- related acute infection. However, increasing numbers of illness are now known to be associated with this particular herpes virus, such as hemophagocytic lymphohistiocytosis (HLH), chronic active EBV infection (CAEBV) and various malignant neoplasms, including nasopharyngeal carcinoma, Hodgikin's disease and gastric cartinoma. It has been suggested by studying these diseases that the cellular targets of vital infection and the modes of the latency of viruses greatly the modify pathogenesis of EBV-related illnesses. EBV-encoded small RNA-1 (EBER1) is expressed in abundance in virtually every cell infected with EBV, regardless of the mode of latency and therefore its expression is often used as a sensitive indicator of EBV association with certain illnesses. In this study, the clinical relevance
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of in situ hybridization flowctyometric analysis (ISH/FCM) of EBER1 expression was evaluated to identify EBV-infected cells quantitatively. Preliminary experiments showed that ISH/FCM using a fluorescence-labeled probe for EBER1 mRNA enabled more quantitative evaluation of the fraction of the infected cells within a sample population than standard ISH using alkaline phosphatase-labeled probe. However, the former was less sensitive when the fraction of the infected cells was smaller than 1 percent. Levels of EBER1 expression were various among different cell lines and the distribution of the fluorescence intensity within a single cell population is relatively wide, indicating that EBER1 expression is controlled by multiple mechanisms. Importantly, the virus-infected cells were easily detectable within peripheral lymphocytes from patients with HLH and CAEBV, but not from normal controls or patients with acute infectious mononucleosis. EBER1 ISH/FCM may serve as a useful tool to monitor sequentially the target cells and the mode of infection in these patients with relative ease. Further information will be available by applying multiparameter analysis of EBER1 expression in combination with different EBV-related gene expression and surface antigen analysis. Less
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