MOLECULAR IMMUNE RESPONSE DETERMINANTS OF RUBELLA VIRUS

风疹病毒的分子免疫反应决定因素

• 批准号: 2063668
• 负责人: Jerry Saul Wolinsky
• 金额: $ 21.95万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-06-01 至 1994-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2063668
• 关键词: Rubivirus; SDS polyacrylamide gel electrophoresis; affinity chromatography; cellular immunity; congenital infection; drug adverse effect; enzyme linked immunosorbent assay; gel electrophoresis; glycoproteins; human tissue; hybridomas; iatrogenic disease; immunofluorescence technique; immunoprecipitation; laboratory mouse; mass spectrometry; molecular pathology; monoclonal antibody; multiple sclerosis; protein sequence; rheumatoid arthritis; rubellas; subacute sclerosing panencephalitis; tissue /cell culture; viral vaccines; virus infection mechanism; virus protein; western blottings

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.

我们的最终目标是确定异常的免疫反应如何 风疹病毒对临床和病理学有贡献 先天性风疹综合征的表现，进行性 风疹全脑炎、疫苗关节痛，或许还有类风湿 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 回应。 这将提供定义所需的数据库 预测的人类免疫反应的改变有助于 风疹的多种并发症。 该项目将定义 诱导风疹病毒所需的最小结构域 中断病毒感染性的抗体反应，以及那些 被加工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。 风疹病毒单克隆抗体库将 扩展并用于定义顺序和构象- 依赖于抗原承载结构域的结构域 多肽。 这些实验将采用新颖的方法 它结合了免疫沉淀、抗体保护的力量 部分蛋白水解消化、SDS-PAGE 和快原子轰击 质谱分析。 体外增殖试验使用 与颗粒硝化纤维素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 合成寡肽将证实这些的重要性 域并提供用于探测关键的试剂 风疹正常反应和异常反应之间的差异 相关综合症。 所获得的知识将增加我们的 了解普通人的正常免疫反应 病原体，提供构建中使用的关键信息 分子工程疫苗，促进其他人的方法 解决这个普遍问题，并提供对异常情况的更好洞察 人类免疫反应及其对病毒持久性的贡献 和病毒诱导的免疫病理学。