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中文摘要
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我们的最终目标是确定异常免疫反应是如何 风疹病毒在临床和病理上的贡献 进行性先天性风疹综合征的临床表现 风疹全脑炎,疫苗关节痛,可能还有类风湿 关节炎和多发性硬化症。一个先决条件是 抗原区分子拓扑图的构建 引起体液和细胞免疫的风疹病毒 回应。这将提供所需的数据库来定义 预测了人类免疫反应的变化,这对 风疹的各种并发症。该项目将定义 风疹病毒诱导所需的最小结构域 中断病毒传染性的抗体反应,以及那些 加工后的结构蛋白的氨基酸序列 并由人类单核细胞呈递以建立细胞免疫。 风疹病毒的单克隆抗体库将是 扩展并用于定义序列和构象- 位于结构上的依赖抗原承载结构域 多肽。这些实验将利用新的方法 它结合了免疫沉淀的力量,抗体受到保护 部分蛋白水解酶、SDS-PAGE和快原子轰击 质谱学。使用的体外增殖试验 与颗粒状硝化棉结合的多肽或其片段 将定义免疫细胞识别的抗原域。已选择 人工合成的寡肽将证实这些 域并提供用于探测危急情况的试剂 风疹中正常反应和异常反应的差异 相关证候。所获得的知识将增加我们的 对这种普通人的正常免疫反应的理解 病原体,提供在构建过程中使用的关键信息 分子工程疫苗,促进了其他疫苗的开发 ,并提供了对异常的更好的洞察力 人类免疫反应及其对病毒持久性的贡献 以及病毒诱导的免疫病理学。
英文摘要
Our ultimate goal is to determine how aberrant immune responses to rubella virus contribute to the clinical and pathological manifestations of the congenital rubella syndrome, progressive rubella panencephalitis, vaccine arthralgia and perhaps rheumatoid arthritis and multiple sclerosis. A prerequisite is the construction of the molecular topography of the antigenic domains of rubella virus which elicit humoral and cell-mediated immune responses. This will provide the required data base to define the predicted alterations in the human immune response which contribute to the diverse complications of rubella. This project will define the minimum domains of rubella virus that are required to induce antibody responses which interrupt virus infectivity, and those amino acid sequences of the structural proteins which are processed and presented by human monocytes to establish cellular immunity. A library of monoclonal antibodies to rubella virus will be expanded and used to define both sequential and conformation- dependant antigen bearing domains residing on the structural polypeptides. These experiments will utilize novel approaches which combine the power of immunoprecipitation, antibody protected partial proteolytic digestion, SDS-PAGE and fast atom bombardment mass spectrometry. In vitro proliferative assays which use polypeptides or their fragments bound to particulate nitrocellulose will define antigenic domains recognized by immune cells. Selected synthetic oligopeptides will confirm the importance of these domains and provide reagents with which to probe critical differences between normal and anomalous responses in rubella related syndromes. The knowledge gained will increase our understanding of normal immune responses to this common human pathogen, provide critical information of use in constructing molecularly engineered vaccines, facilitate the approach of others to this general problem, and provide better insight into aberrant human immune responses and their contribution to viral persistence and virus-induced immunopathology.
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