3RD VARIABLE DOMAIN OF THE HIV-1 ENVELOPE

HIV-1 包膜的第三个可变域

• 批准号: 3145424
• 负责人: THOMAS J MATTHEWS
• 金额: $ 21.39万
• 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 1995-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3145424
• 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：(改编自申请者的摘要)一个已经证明 对抗体阻断淋巴细胞感染特别敏感 在HIV包膜糖蛋白的第三个可变区(V3)环中 (GP)120。这里提出的研究计划就是针对这一点 为了了解HIV包膜的结构和结构 功能以及观察到的氨基酸序列在这个过程中的变化 结构域影响感染性和抗体反应性。这项工作 将扩展到原代单核/巨噬细胞的感染 到T细胞系，因为附着、穿透和作用的细节 这两个谱系的细胞中的抗体可能不同。主要问题 解决的问题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、增强或无效 原代单核/巨噬细胞感染的研究。第二， V3结构域的结构/功能未知，但与 膜蛋白水解酶已经被提出，这种可能性将得到检验。 第三，对克隆的HXB2分离株进行定点突变 用于识别该变量中的序列和结构特征 对病毒传染性最关键的领域。在这些步骤中 被阻止的非传染性病毒构造将使用 猴病毒40(SV40)为载体的突变体表达与研究 在没有其他HIV组分的情况下，包膜糖蛋白。最后， 第三个目的是复制，制备致突变的艾滋病毒 将在中和试验中研究是否合格，以确定残基位置 它们对中和抗体的影响最大。