Broad Neutralizing Monoclonal Antibodies From HIV Controllers

描述（由申请人提供）：该项目的长期目标是鉴定广泛识别HIV-1包膜糖蛋白（Env）并在体外阻断感染的新型单克隆抗体（mab），以指导疫苗开发。这一目标将在一组HIV-1感染者中实现，这些人在没有抗逆转录病毒治疗（天然病毒抑制剂/NVS）的情况下控制了感染，并且有循环广泛中和抗体（broad nab）。我们方法的一个关键要素是开发一种新的测定方法来普查env特异性记忆B细胞克隆（BMem），这使得快速和直接克隆全长单克隆抗体（mab）成为可能。这些单克隆抗体将具有表位特异性和中和宽度的特征，以创建在HIV-1感染控制过程中产生的BMem的克隆谱。该信息将用于验证这样的假设，即中和广度是由由一系列中和特异性组成的多克隆反应决定的，而不是由一个或极少数中和特异性组成的少克隆反应。验证这一假设是我们确定广泛识别Env并阻断体外感染的新型单克隆抗体以指导针对HIV-1的疫苗开发的长期目标的关键。有两个具体目标。目的1-从具有持续广泛中和抗体反应的NVS中开发env特异性BMem的克隆特异性谱-抗env反应的克隆特异性谱将通过限制稀释分析、单抗分离和表位定位来确定，以确定不同env表位特异性BMem克隆的相对优势。目的-2-比较血浆抗体和单克隆抗体之间的中和宽度，以确定必须汇集的单克隆抗体数量，以重建循环抗体池的中和宽度。该数据将用于确定正在进行的广泛nAb反应的克隆性。这一目的将完成对中和广度由由一系列中和特异性组成的多克隆反应决定的假设的检验，而不是由一个或极少数中和特异性组成的少克隆反应。目前还没有预防艾滋病的疫苗。本应用程序中提出的工作将调查一些人如何在没有抗逆转录病毒药物治疗的情况下控制HIV-1感染多年。这一信息应该对研制艾滋病疫苗有用。

DESCRIPTION (provided by applicant): The long-term goal of this project is to identify novel monoclonal antibodies (mAbs) that broadly recognize the HIV-1 envelope glycoprotein (Env) and block infection in vitro to guide vaccine development. This goal will be pursued in a cohort of HIV-1 infected individuals who control their infections in the absence of anti-retroviral therapy (Natural Virus Suppressors/NVS) and who have circulating broadly neutralizing antibodies (broad nAbs). A key element of our approach is the development of a new assay to census Env-specific memory B cell clones (BMem) that allows the rapid and direct cloning of full-length monoclonal antibodies (mAbs). These mAbs will be characterized for epitope specificity and neutralization breadth to create clonal profiles of the BMem that are generated during the control of HIV-1 infection. This information will be used to test the hypothesis that neutralization breadth is determined by a polyclonal response comprised of a mosaic of neutralizing specificities as opposed to a pauciclonal response comprised of one or a very few neutralizing specificities. Testing this hypothesis is key to our long-term goal of identifying novel mAbs that broadly recognize Env and block infection in vitro to guide vaccine development against HIV-1. There are two specific aims. Aim 1- To develop clonal specificity profiles of Env-specific BMem from NVS who have ongoing broadly neutralizing antibody responses- Clonal specificity profiles of anti-Env responses will be determined by limiting dilution analysis, mAb isolation, and epitope mapping to determine the relative dominance of BMem clones specific for different Env-epitopes. Aim-2- To compare neutralization breadth between plasma antibodies and mAbs representing a full clonal profile of BMem to determine the number of mAbs that must be pooled to reconstruct the neutralization breadth of the circulating antibody pool. This data will be used to determine the clonality of an ongoing broad nAb response. This aim will complete testing the hypothesis that neutralization breadth is determined by a polyclonal response comprised of a mosaic of neutralizing specificities as opposed to a pauciclonal response comprised of one or a very few neutralizing specificities. Currently there is no vaccine against AIDS. The work proposed in this application will investigate how some people control HIV-1 infection for many years without anti- retroviral drug therapy. This information should be useful in making a vaccine against AIDS.