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

使用具有定义的免疫缺陷的动物，例如SCID（严重的合并免疫缺陷）小鼠和无静脉裸鼠，研究了免疫功能低下的宿主中单纯疱疹病毒感染的发病机理。这项研究主要集中于潜伏/重新激活，尽管发展了几种有效的抗病毒药，但尽管发展了病毒的独特病理机制以及未解决的临床问题。通过该赠款获得的流式细胞仪，分析了从血液，脾，区域淋巴结和炎症组织（例如角膜和视网膜）获得的免疫病理和免疫病理效应细胞。体外和体内耗竭的淋巴细胞特定子集也用于识别免疫病理学和/或免疫防御的负责任子集。将寡核苷酸与该赠款获得的DNA合成剂合成，并用作聚合酶链反应（PCR）的引物或探针，然后与DOT印迹和/或Southern blot杂交进行了RNA PCR。通过PCR检测到没有症状的唾液中的单纯疱疹病毒，并将PCR与传统病毒分离的阳性样品的敏感性和临床意义进行了比较。人类巨细胞病毒DNA，另一种疱疹病毒，并且在患者血液和其他组织中也很难在体外生长。从感染的细胞和动物组织中获得了DNA和RNA，并分析了各种病毒基因组DNA和/或转录物RNA.RNA的simples，来自潜在感染的小鼠三叉神经节具有和没有不同的重新激活刺激的小鼠三叉神经节，并通过发现其他仅检测ICP0 rNA或其他病毒转录的反应激活模式。分子分析其包膜糖蛋白C基因或胸苷激酶基因的临床病毒分离株。亲本病毒和重度的致病性