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Virological and molecular biological studies on human herpsvirus 6 (HHV-6).

人类疱疹病毒 6 (HHV-6) 的病毒学和分子生物学研究。

基本信息

批准号：
01440032
负责人：
YAMANISHI Koichi
金额：
$ 12.99万
依托单位：
Osaka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (A)
财政年份：
1989
资助国家：
日本
起止时间：
1989 至 1991
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-01440032/
关键词：
HHV-6 Exanthem Subitum PCR DNA Cloning Latent Infection HHVー6 DNAクロ-ニング

项目摘要

1.Establishment of titration system and neutralizing system of HHV-6: We have been using peripheral blood lymphocytes to titrate virus. Recently we could develop a new system by using MT-4 cells, which is a established cell line, for titration of virus and for neutralizing test. Since this system is useful for not only titrate virus but also antibody test, seroepidemiology in Japan and Thailand were attempted.2.Analysis of HHV-6 DNA by cloning and sequence: DNA cloning and its sequences of HHV-6 have been done since 1989. Since size of DNA is so big, we could not complete yet, but we are identifying some of them. On the way of this process, the system of detection of HHV-6 DNA by polymerase chain reaction (PCR) has been developed, and we have used this system for detection of HHV-6 DNA in the latent system and for detection of DNA in various clinical samples such as lymphoma, chronic fatique syndrome.3.Analysis of polypeptide of HHV-6: The virion of HHV-6 were purified and analyzed. More than 29 polypeptide including 6 glycoproteins were identified. After collection of monoclonal antibodies to glycoproteins of HHV-6, 3 glyproteins were characterized by biological activity and processing of them in infected cell. Furthermore, it was found HHV-6 induce virus specific DNA polymerase, but not thymidine kinase.4.Development of animal system for HHV-6 infection: Although we could not find any animal system in small' rodents, we could detect antibodies to HHV-6 in various monkeys. Recently we could find cynomolgus and African green monkeys can be infected with HHV-6, and this system will be useful for the system of latent infection and analysis of pathogenesis of HHV-6 in future

1.HHV-6滴定体系和中和体系的建立：我们一直采用外周血淋巴细胞滴定病毒。最近我们可以利用MT-4细胞（一种已建立的细胞系）开发一个新系统，用于病毒滴定和中和测试。由于该系统不仅可用于滴定病毒，而且还可用于抗体检测，因此在日本和泰国进行了血清流行病学尝试。2.通过克隆和序列分析HHV-6 DNA：HHV-6的DNA克隆和序列分析自1989年以来一直在进行。由于DNA的大小如此之大，我们还无法完成，但我们正在识别其中的一些。在此过程中，我们开发了聚合酶链式反应（PCR）检测HHV-6 DNA的系统，并利用该系统检测潜伏系统中的HHV-6 DNA，以及检测淋巴瘤、慢性疲劳综合征等各种临床样本中的DNA。 3.HHV-6多肽分析：对HHV-6病毒体进行纯化和分析。鉴定出超过29种多肽，包括6种糖蛋白。收集 HHV-6 糖蛋白的单克隆抗体后，对 3 种糖蛋白的生物活性及其在感染细胞中的加工进行了表征。此外，还发现HHV-6诱导病毒特异性DNA聚合酶，但不诱导胸苷激酶。4.HHV-6感染动物系统的开发：虽然我们在小型啮齿动物中没有找到任何动物系统，但我们可以在各种猴子中检测到HHV-6抗体。最近我们发现食蟹猴和非洲绿猴可以感染HHV-6，该系统对于将来HHV-6潜伏感染系统和发病机制分析具有重要意义。