Study of herpes simplex virus latency and the host defense mechanism against the infection

单纯疱疹病毒潜伏期及宿主防御机制的研究

基本信息

批准号：
03404025
负责人：
MORI Ryoichi
金额：
$ 14.21万
依托单位：
KYUSHU 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：
1991
资助国家：
日本
起止时间：
1991 至 1993
项目状态：
已结题

项目摘要

Pathogenesis of herpes simplex virus infection in immunocompromised hosts were studied using animals with defined immunodeficiency, e.g., SCID (severe combined immunodeficiency) mice and athymic nude mice. This study was mainly focused on latency/reactivation, the unique pathologic mechanisms of the virus and an unsolved clinical problem despite the development of several potent antiviral drugs. Immunological and immunopathological effector cells obtained from the blood, spleen, regional lymph nodes and the inflammatory tissues such as cornea and retina, were analyzed for their surface markers with the flow cytometer obtained by this grant. In vitro and in vivo depletion of a specific subset of the lymphocytes were also used to identify the responsible subset for the immunopathology and/or immunological defense. Oligonucleotides were synthesized with the DNA synthesizer obtained by this grant and used as primers or probes for polymerase chain reaction (PCR) and RNA PCR followed with dot blot and/or Southern blot hybridization. Herpes simplex virus secreted into saliva without symptoms were detected by PCR, and PCR was compared with the conventional virus isolation for its sensitivity and clinical significance of the positive samples. Human cytomegalovirus DNA, another herpesvirus and is difficult to grow in vitro was also detected by PCR from patient blood and other tissues. Both DNA and RNA were obtained from infected cells and animal tissues and were analyzed for various viral genome DNA and/or transcript RNA.RNA simples from latently infected mouse trigeminal ganglia with and without different reactivation stimulus were studied and different reactivation patterns characterized by the detection of ICP0 RNA only or detection of other viral transcripts were reported. Clinical virus isolates with distinct pathogenicity were molecularly analyzed for their envelope glycoprotein C genes or thymidine kinase genes. Pathogenicity of both the parent virus and the recombina

使用具有明确免疫缺陷的动物，例如 SCID（严重联合免疫缺陷）小鼠和无胸腺裸鼠，研究了免疫受损宿主中单纯疱疹病毒感染的发病机制。这项研究主要集中在潜伏/重新激活、病毒独特的病理机制以及尽管开发了几种有效的抗病毒药物但尚未解决的临床问题。使用本次资助获得的流式细胞仪分析从血液、脾脏、区域淋巴结以及角膜和视网膜等炎症组织中获得的免疫学和免疫病理学效应细胞的表面标记。淋巴细胞特定子集的体外和体内耗竭也用于鉴定免疫病理学和/或免疫防御的负责子集。使用本次资助获得的DNA合成仪合成寡核苷酸，并用作聚合酶链反应（PCR）和RNA PCR的引物或探针，然后进行斑点印迹和/或Southern印迹杂交。采用PCR方法检测分泌到唾液中的无症状单纯疱疹病毒，并与常规病毒分离方法比较其阳性样本的敏感性和临床意义。人类巨细胞病毒 DNA（另一种很难在体外生长的疱疹病毒）也通过 PCR 从患者血液和其他组织中检测到。从受感染的细胞和动物组织中获得 DNA 和 RNA，并分析各种病毒基因组 DNA 和/或转录本 RNA。研究了来自潜伏感染的小鼠三叉神经节的 RNA 简单物，有和没有不同的重新激活刺激，并通过检测表征了不同的重新激活模式据报道，仅检测 ICP0 RNA 或检测其他病毒转录本。对具有不同致病性的临床病毒分离株的包膜糖蛋白 C 基因或胸苷激酶基因进行了分子分析。亲本病毒和重组病毒的致病性