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中文摘要
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我们的最终目标是确定异常的免疫反应是如何对 风疹病毒有助于临床和病理 进行性先天性风疹综合征的表现 风疹全脑炎,疫苗关节痛,也许还有类风湿 关节炎和多发性硬化症 一个先决条件是 抗原结构域的分子拓扑结构的构建 风疹病毒引起体液和细胞介导的免疫 应答 这将提供所需的数据库, 预测的人类免疫反应的变化, 风疹的各种并发症。 该项目将定义 风疹病毒的最小结构域, 阻断病毒传染性的抗体反应, 被加工的结构蛋白的氨基酸序列 并由人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 扩展并用于定义序列和构象- 依赖性抗原承载结构域驻留在结构上 多肽。 这些实验将利用新的方法 它联合收割机了免疫沉淀的力量, 部分蛋白酶切、SDS-PAGE和快原子轰击 质谱分析法来 体外增殖测定,其使用 多肽或其片段与颗粒状硝化纤维素结合 将定义免疫细胞识别的抗原域。 选择 合成寡肽将证实这些的重要性, 结构域,并提供用于探测关键 风疹正常反应与异常反应的差异 相关综合征。 获得的知识将增加我们的 了解对这种常见人类的正常免疫反应, 病原体,提供关键信息,用于构建 分子工程疫苗, 这个普遍的问题,并提供更好的洞察异常 人类免疫应答及其对病毒持续存在的贡献 和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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