PRODUCTION OF HUMAN MONOCLONAL ANTIBODIES TO INFLUENZA V

人类 V 型流感单克隆抗体的生产

• 批准号: 3135910
• 负责人: ARTHUR L FRANK
• 金额: $ 11.29万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 1985
• 资助国家: 美国
• 起止时间: 1985-09-30 至 1988-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3135910
• 关键词: B lymphocyte; Epstein Barr virus; antibody specificity; child (0-11); enzyme linked immunosorbent assay; immunoglobulin G; microorganism immunology; monoclonal antibody; radioimmunoassay; tissue /cell culture; virus antigen

The goal of the proposed research is to produce human monoclonal antibodies with specificity for influenza virus antigens and to define their immunologic and biologic properties. Peripheral blood B lymphocytes from persons with recent exposure to influenza antigens will be infected with Epstein-Barr virus to produce continuous lymphoblastoid cell lines. Tissue culture fluids will be screened for influenza antibody by an ELISA technique that utilizes purified influenza virus and enzyme conjugated antihuman IgG antibodies. Secreting cell lines will be grown in cloning conditions and the antibody will be harvested at all stages of growth. Antibody specificity will be tested by ELISA, hemagglutination-inhibition and microneutralization tests using strains representative of major variants within a subtype. They will also be tested for their ability to inhibit binding of representative mouse monoclonal antibodies to each of the four proposed antigenic sites on the hemagglutinin molecule. Comparisons with mouse antibodies will determine if both species identify similar antigenic structures on the viral hemagglutinin. Such information is essential for interpreting the relevance of studies of antigenic variation currently performed with the mouse reagents. This antigenic variation, or "drift," of influenza virus is an important factor in the yearly outbreaks of human disease. The relative avidity/affinity of antihemagglutinin human monoclonal antibodies will be determined by competition radioimmunoprecipitation assays and, in combination with the antibody concentration, and IgG subclass, will be used to compare the efficiencies with which the antibodies inhibit viral hemagglutination and neutralize virus infectivity. This is important for understanding the biologic tests used in assessments of immunity to influenza and may provide information on vaccine formulation so that optimal protective responses will be induced. Possible antigenic differences between egg-grown and tissue culture-grown virus will be sought with human monoclonal antibodies. In the future human monoclonal antibodies may serve as laboratory reagents for analysis of influenza strains or standardization of serological tests and eventually they may be candidates for human therapeutic use.

这项研究的目标是生产人类单克隆抗体 对流感病毒抗原具有特异性， 免疫学和生物学特性。 外周血B淋巴细胞 最近接触过流感抗原的人会感染 EB病毒产生连续的淋巴母细胞系。 组织 将通过ELISA筛选培养液中的流感抗体 利用纯化的流感病毒和酶结合的技术 抗人IgG抗体。 分泌细胞系将克隆生长 在所有生长阶段收获抗体。 将通过ELISA、血凝抑制试验检测抗体特异性 和使用主要微生物代表菌株的微量中和试验 一个子类型中的变体。 他们也将被测试他们的能力， 抑制代表性小鼠单克隆抗体与 血凝素分子上的四个抗原位点。 与小鼠抗体的比较将确定这两个物种是否能够识别 病毒血凝素上的抗原结构相似。 此类信息 对于解释抗原性研究的相关性至关重要， 目前使用小鼠试剂进行的变异。 该抗原 流感病毒的变异或“漂移”是流感病毒感染的一个重要因素。 每年人类疾病的爆发。 的相对亲合力/亲和力 将通过以下方法测定抗血凝素人单克隆抗体： 竞争性放射免疫沉淀试验，并与 抗体浓度和IgG亚类，将用于比较 抗体抑制病毒血凝的效率， 中和病毒的传染性 这对于理解 用于评估流感免疫力的生物学检测， 关于疫苗配方信息， 将被诱导。 鸡胚和鸡胚之间可能存在的抗原差异 将用人单克隆抗体寻找组织培养生长的病毒 抗体的 在未来，人类单克隆抗体可以作为 流感病毒株分析或标准化用实验室试剂 血清学测试，最终它们可能是人类的候选者。 治疗用途。