HUMAN ANTIBODIES TO HIV-1 BY REPERTOIRE CLONING

通过全库克隆获得 HIV-1 人类抗体

• 批准号: 6945503
• 负责人: Dennis R. Burton
• 金额: $ 50.53万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-07-01 至 2010-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6945503
• 关键词: AIDS; AIDS vaccines; B lymphocyte; antiAIDS agent; antibody neutralization test; antigen antibody reaction; antiviral antibody; clinical research; genetic techniques; human genetic material tag; human immunodeficiency virus 1; human subject; human tissue; molecular cloning; monoclonal antibody; neutralizing antibody; passive immunization; patient oriented research; phage display; vaccine development

DESCRIPTION (provided by applicant): The current consensus is that a successful HIV vaccine will need to elicit both neutralizing antibody and cellular immune responses. The antibody response should be broadly neutralizing. The feasibility of eliciting such a response is suggested by the existence of a small panel of broadly neutralizing human monoclonal antibodies (mAbs). These mAbs are valuable tools in vaccine design. They help to define conditions for antibody-mediated protection, they reveal neutralizing epitopes, they provide quality control for vaccine candidates (on which neutralizing epitopes should be expressed) and they lead directly to structures that are vaccine candidates e.g. engineered envelope molecules, peptides and carbohydrates. However, the number of broadly neutralizing mAbs is small and they are all derived from subtype B-infected donors. In order to have a larger panel of mAbs that will more effectively neutralize global HIV-1 isolates and that will stimulate vaccine research, we propose a renewed effort to generate such Abs using phage display, yeast display and memory B cell technologies. This effort is timely given a number of advances including the identification of donors with broad serum neutralizing activity and the refinement of technologies for human mAb generation and selection. Aim 1 is to generate and characterize broadly neutralizing human mAbs from subtype A, B, and C-infected donors. 4E10 is clearly the most broadly neutralizing mAb described to date but its potency is generally only moderate. We have recently solved the structure of Fab 4E10 in complex with its peptide antigen and now propose to apply in vitro evolution techniques to improve the affinity and neutralizing characteristics of 4E10. Aim 2 is to generate affinity-enhanced versions of 4E10 for use as entry inhibitors and as improved templates for immunogen design. Both aims are focused on providing the tools that will eventually allow us to generate or design immunogens that reliably elicit broadly neutralizing Abs.

描述（由申请人提供）：当前共识是成功的HIV疫苗需要引起中和抗体和细胞免疫反应。抗体反应应广泛中和。引起这种反应的可行性是由一小部分中和人类单克隆抗体（mAb）的存在提出的。这些mAB是疫苗设计中的宝贵工具。它们有助于定义抗体介导的保护条件，揭示了中和表位，它们为候选疫苗提供了质量控制（应在其上表达中和表位），并直接导致候选疫苗的结构。设计的包膜分子，肽和碳水化合物。但是，广泛中和的mAb的数量很少，它们都来自亚型B感染者。为了拥有更大的mAB面板，可以更有效地中和全局HIV-1分离株并刺激疫苗研究，我们提出了一种新的努力，以使用噬菌体显示，酵母显示和记忆B细胞技术来生成此类ABS。考虑到许多进步，包括确定具有广泛的血清中和活性的捐赠者以及人类MAB生成和选择的技术的完善。目的1是从亚型A，B和C感染的供体中产生和表征广泛中和的人物BAB。 显然，4E10是迄今为止描述的最广泛中和的MAB，但其效力通常只是中和。我们最近已经解决了与其肽抗原复合物中Fab 4e10的结构，现在建议应用体外进化技术来改善4E10的亲和力和中和特性。 AIM 2是生成4E10的亲和力增强版本，以用作入口抑制剂，并用作改进的免疫原模板。 两种目标都集中在提供最终使我们能够生成或设计免疫原的工具，从而可靠地引起广泛中和ABS。