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: 3135911
• 负责人: ARTHUR L FRANK
• 金额: $ 12.86万
• 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/3135911
• 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.

这项拟议的研究的目标是生产人类的单抗。 具有流感病毒抗原的特异性，并确定其 免疫学和生物学特性。中国人外周血中B淋巴细胞 最近接触过流感抗原的人会感染 爱泼斯坦-巴尔病毒可产生连续的淋巴母细胞系。组织 培养液将通过酶联免疫吸附试验检测流感抗体。 利用纯化的流感病毒和偶联酶的技术 抗人免疫球蛋白抗体。分泌细胞系将在克隆中生长 条件和抗体将收获在生长的所有阶段。 抗体特异性检测采用酶联免疫吸附试验、血凝抑制试验 用代表主要毒株的菌株进行微中和试验 子类型中的变体。他们还将接受测试，以确定他们是否有能力 抑制代表性的鼠单抗与每一种 血凝素分子上的四个建议的抗原点。 与老鼠抗体的比较将确定这两个物种是否 病毒血凝素的抗原结构相似。这样的信息 对于解释抗原性研究的相关性是必不可少的 目前使用的是老鼠试剂的变异。这种抗原性 流感病毒的变异或“漂移”是导致 每年爆发的人类疾病。相对亲和力/亲和力 抗血凝素人源单抗将通过 竞争放射免疫沉淀法，并结合 抗体浓度和免疫球蛋白亚类将用于比较 抗体抑制病毒血凝和 中和病毒的传染性。这对于理解 用于评估对流感的免疫力的生物测试，并可提供 关于疫苗配方的信息，以便获得最佳保护反应 将会被诱导。蛋鸡和鸡的抗原性可能存在差异 组织培养培养的病毒将与人的单抗一起寻找 抗体。在未来，人类单抗可能会作为 流感病毒株分析或标准化的实验室试剂 血清学测试，最终它们可能成为人类的候选 治疗用途。