HIV GP120/GP41 ANALYSIS USING MONOCLONAL ANTIBODIES

这一建议所依据的主要假设是， 已知的表位结构的包膜蛋白的抗原决定簇， HIV病毒和人类淋巴细胞T4受体将使我们能够 了解这些蛋白质的功能位点 其参与gp 120-T4受体相互作用。 等 知识将导致能够防止初始 感染和病毒传播。 为了验证这一 假设我们提出以下三个具体目标。 1)我们 建议描述HIV包膜的表位结构 蛋白质和人T4受体的开发和定义， 功能和结构水平的一系列鼠单克隆抗体 包膜蛋白和人T4受体的抗体。 2)我们建议开发人-鼠嵌合抗体 靶向选定的“功能性”GP 120表位。 搜索将 这些抗体将优先结合到 通常不被HIV免疫系统识别的表位 感染的人。 这些嵌合抗体将被测试， 它们与表达HIV包膜蛋白的细胞结合的能力， 结合到表面后被内吞，并介导 在存在下表达包膜蛋白的细胞的细胞溶解 人类补体 3）。我们建议开发一系列的反- 抗鼠单克隆抗体的独特型抗体， 识别“功能上”重要的gp 120表位， 表位识别人T4的单克隆抗体 淋巴细胞受体 这些抗独特型抗体将 通过序列分析和分子模拟研究， 他们独特的三维结构。 以这种方式， 我们计划确定功能表位的空间结构， 包膜蛋白和T4受体。 除了 通过制定这些具体目标，本提案将作为 单克隆抗体生产设施 本国家标准项目1、2、3、4、5和7中的研究试剂 艾滋病药物发现合作小组。

The principle hypothesis upon which this proposal is based is that knowledge of the epitope structure of the envelope protein of the HIV virus and the human lymphocyte T4 receptor will allow us to develop an understanding of the functional sites on these proteins which are involved in gp 120-T4 receptor interaction. Such knowledge will lead to modalities capable of preventing initial infection and spread of virus in established infection. To test this hypothesis we propose the following three specific aims. 1) We propose to characterize the epitope structure of the HIV envelope protein and the human T4 receptor by developing and defining at a functional and structural level a series of murine monoclonal antibodies to the envelope protein and to the human T4 receptor. 2) We propose to develop human-murine chimeric antibodies targeted for selected "functional" gp120 epitopes. A search would be made for such antibodies which would preferentially bind to epitopes not normally recognized by the immune system of HIV infected individuals. These chimeric antibodies will be tested for their ability to bind to cells expressing the HIV envelope protein, to be endocytosed after binding to the surface, and to mediate cytolysis of cells expressing envelope protein in the presence of human complement. 3). We propose to develop a series of anti- idiotype antibodies to murine monoclonal antibodies which recognize "functionally" important gp 120 epitopes and to monoclonal antibodies whose epitopes recognize the human T4 lymphocyte receptor. These anti-idiotypes antibodies will be studied by sequence analysis and molecular modeling to determine the three dimensional structure of their idiotopes. In this manner, we plan to define the steric structure of functional epitopes on the envelope protein and the T4 receptor. In addition to developing these specific aims, this proposal will serve as a core monoclonal antibody facility for the production of monoclonal reagents for studies in programs 1, 2, 3, 4, 5, ad 7 of this National Cooperative Drug Discovery Group for AIDS.