TOPOLOGICAL ANALYSIS OF HIV-1 ENVELOPE GLYCOPROTEINS

HIV-1 包膜糖蛋白的拓扑分析

• 批准号: 6148301
• 负责人: JOSEPH G SODROSKI
• 金额: $ 30.64万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-05-01 至 2005-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6148301
• 关键词: HIV envelope protein gp120; HIV envelope protein gp160; glycoprotein structure; human immunodeficiency virus 1; neutralizing antibody

DESCRIPTION (Adapted from Applicant's Abstract): The basic goal of this proposal is to understand the structural differences that exist between two highly related forms of the HIV glycoprotein that differ markedly in neutralization sensitivity through the use of chimeric env genes and antibody probes. The authors argue that the development of an effective and safe human immunodeficiency virus (HIV-1) vaccine would benefit greatly from an understanding of protective immune responses. In several animal models, including the infection of macaques with simian-human immunodeficiency viruses (SHIVs), neutralizing antibodies against the challenge virus can mediate protection. However, the applicants point out that two difficulties face the practical utilization of neutralizing antibodies for HIV-1 prophylaxis: a) the diversity of the HIV-1 envelope glycoproteins, the major target for neutralizing antibodies; and b) the relative resistance of primary HIV-1 isolates, compared with laboratory-adapted virus strains, to neutralization by antibodies. Although the major variable loops of the gp120 external envelope glycoprotein are known to contribute to these properties, understanding of the structure of these determinants lags behind that of more conserved envelope glycoprotein components. Results from the PI's laboratory have shown that in vivo passage of a SHIV bearing the envelope glycoproteins of a laboratory-adapted HIV-1 isolate, HXBc2, resulted in a virus that caused rapid CD4-positive T-lymphocyte depletion and AIDS in rhesus monkeys. A molecularly cloned virus, SHIV-HXBc2P 3.2, which contains the HIV-1 envelope glycoproteins of the passaged virus, was shown to be pathogenic in monkeys. The HXBc2P 3.2 envelope glycoproteins were markedly resistant to neutralization by soluble CD4 and several antibodies compared with the parental HXBc2 envelope glycoproteins. Thus, in vivo passage resulted in the acquisition of neutralization resistance typical of that of primary HIV-1 isolates.The specific aims of this proposal are: 1. To create recombinants between the neutralization-sensitive HXBc2 and the neutralization-resistant HXBc2P 3.2 envelope glycoproteins to map the genetic determinants of resistance to neutralization by various antibodies. 2. To compare the structures of the parental and recombinant gp120 glycoprotein monomers by antibody cross-competition analysis. To study the structure of HIV-1 envelope glycoprotein trimers by antibody cross-competition analysis, and to characterize differences between HXBc2 and HXBc2P 3.2 envelope glycoprotein trimers.

描述（改编自申请人的摘要）：此的基本目标 建议是了解两个之间存在的结构差异 HIV糖蛋白的高度相关形式在 通过使用嵌合ENV基因和抗体中和敏感性 探针。作者认为，有效和安全的人的发展 免疫缺陷病毒（HIV-1）疫苗将从中受益匪浅 了解保护性免疫反应。在几种动物模型中 包括猕猴感染猿猴免疫缺陷病毒 （SHIVS），对挑战病毒的中和抗体可以介导 保护。但是，申请人指出，两个困难面临 实际利用中和HIV-1预防的抗体：a） HIV-1信封糖蛋白的多样性，这是 中和抗体； b）原代HIV-1的相对电阻 与实验室适应的病毒菌株相比，分离株与 抗体。虽然GP120外部信封的主要变量循环 已知糖蛋白有助于这些特性，了解 这些决定因素的结构滞后于更为保守的信封的结构 糖蛋白成分。 PI实验室的结果表明 带有信封糖蛋白的湿因的体内通过 实验室适应的HIV-1分离株HXBC2导致病毒迅速 CD4阳性的T淋巴细胞耗竭和恒河猴的辅助。分子 克隆病毒，Shiv-HxBC2P 3.2，其中包含HIV-1信封糖蛋白 在猴子中证明了传递的病毒是致病性的。 HXBC2P 3.2 包膜糖蛋白通过可溶性CD4明显抵抗中和 与亲本HXBC2包膜糖蛋白相比，几种抗体。 因此，体内通道导致了中和抗性的获取 本提案的特定目的 为：1。在中和敏感的HXBC2和 耐中和的HXBC2P 3.2包膜糖蛋白绘制 各种抗体抵抗中和的遗传决定因素。 2。 比较父母和重组GP120糖蛋白的结构 单体通过抗体交叉竞争分析。研究结构 HIV-1包膜糖蛋白三聚体通过抗体交叉竞争分析和 表征HXBC2和HXBC2P 3.2包膜糖蛋白之间的差异 三线。