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

使用具有确定的免疫缺陷的动物，例如，SCID（严重联合免疫缺陷）小鼠和无胸腺裸鼠。这项研究主要集中在潜伏/再激活，病毒的独特病理机制和一个尚未解决的临床问题，尽管开发了几种有效的抗病毒药物。从血液、脾脏、局部淋巴结和炎性组织如角膜和视网膜中获得的免疫和免疫病理效应细胞用本基金获得的流式细胞仪分析其表面标志物。淋巴细胞特定亚群的体外和体内耗竭也用于鉴定免疫病理学和/或免疫防御的负责亚群。用本授权获得的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方法检测无症状分泌到唾液中的单纯疱疹病毒，并与常规病毒分离方法比较其敏感性及阳性标本的临床意义。人巨细胞病毒DNA，另一种疱疹病毒，难以在体外生长，也通过PCR从患者血液和其他组织中检测到。从感染的细胞和动物组织中获得DNA和RNA，并分析了各种病毒基因组DNA和/或转录物RNA.RNA样品从潜伏感染的小鼠三叉神经节与无不同的再激活刺激进行了研究和不同的再激活模式，其特征在于由ICP 0 RNA检测或其他病毒转录物的检测报告。对临床分离的致病性不同的病毒进行了包膜糖蛋白C基因和胸苷激酶基因的分子分析。亲本病毒和重组病毒的致病性

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