3RD VARIABLE DOMAIN OF THE HIV-1 ENVELOPE

HIV-1 包膜的第三个可变域

• 批准号: 3145425
• 负责人: THOMAS J MATTHEWS
• 金额: $ 21.02万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-09-30 至 1995-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3145425
• 关键词: AIDS; antibody neutralization test; antiviral antibody; human immunodeficiency virus 1; macrophage; membrane proteins; monocyte; peptidases; protein structure function; site directed mutagenesis; virus envelope; virus protein

DESCRIPTION: (Adapted from applicant's abstract) A site that has proven particularly sensitive to antibody blockade of lymphocyte infection falls in the third variable domain (V3) loop of the HIV envelope glycoprotein (gp) 120. The research program proposed here is directed solely at this region of the HIV envelope in an effort to understand its structure and function and how the observed amino acid sequence variation within this domain influences infectivity as well as antibody reactivity. The work will be extended to infection of primary monocytes/macrophages in addition to T cell lines since details of attachment, penetration, and role of antibody in cells of these two lineages may differ. The major questions addressed and approaches followed are four fold. First, the investigator proposes to test if antibodies to V3 neutralize, enhance, or have no effect on infection of primary monocytes/macrophages. Second, the structure/function of the V3 domain is unknown but interaction with a membrane protease has been proposed and this possibility will be tested. Third, site-directed mutagenesis of the HXB2 cloned isolate will be utilized to identify sequence and structural features within this variable domain most critical for virus infectivity. The steps at which the non-infectious virus constructs are blocked will be characterized using a simian virus 40 (SV40)-based vector to express and study the mutant envelope glycoprotein in the absence of other HIV components. Finally, mutagenized HIV prepared in the third aim above which are replication competent will be studied in neutralization assays to define residue sites which have the greatest impact on neutralizing antibody.

描述：（改编自申请人的摘要）一个网站，已证明 对抗体阻断特别敏感的淋巴细胞感染福尔斯 在HIV包膜糖蛋白的第三可变结构域（V3）环中 (gp)一百二十 这里提出的研究计划仅针对这一点 区域的艾滋病毒信封，以了解其结构和 功能以及观察到的氨基酸序列变异如何在此 结构域影响感染性以及抗体反应性。 工作 将扩展到感染初级单核细胞/巨噬细胞， 由于T细胞系的附着，渗透和作用的细节， 这两种谱系的细胞中的抗体可能不同。 的主要问题 处理和遵循的方法有四个方面。 首先，调查员 建议测试V3抗体是否中和，增强或没有影响 对原代单核细胞/巨噬细胞的感染。 二是 V3结构域的结构/功能是未知的，但与 已经提出了膜蛋白酶，并且将测试这种可能性。 第三，将HXB 2克隆分离物的定点诱变进行了研究。 用于鉴定该变量内的序列和结构特征 对病毒传染性最关键的领域。 的步骤， 非感染性病毒构建体被阻断的特征将使用 猴病毒40（SV 40）为基础的载体表达和研究的突变体 包膜糖蛋白在缺乏其他HIV组分的情况下。 最后， 在上述第三个目的中制备的诱变的HIV， 将在中和试验中对活性进行研究，以确定残留位点 对中和抗体的影响最大。