Broad Neutralizing Monoclonal Antibodies From HIV Controllers

描述(申请人提供)：该项目的长期目标是鉴定新的能广泛识别HIV-1包膜糖蛋白(Env)并在体外阻断感染的单抗，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(天然病毒抑制者/NV)的情况下控制自己的感染，并具有广泛的中和抗体(广泛的NAB)。我们方法的一个关键因素是开发一种新的方法来普查环境特异性记忆B细胞克隆(BMem)，该方法允许快速和直接克隆全长单抗(MAbs)。这些单抗将以表位特异性和中和广度为特征，以创建在控制HIV-1感染期间产生的BMem的克隆图谱。这些信息将被用来检验这样的假设，即中和广度由由中和特异性马赛克组成的多克隆反应决定，而不是由一个或极少数中和特异性组成的稀有克隆反应决定。验证这一假设是我们长期目标的关键，我们的长期目标是识别广泛识别环境病毒并在体外阻断感染的新型单抗，以指导针对HIV-1的疫苗开发。有两个具体目标。目的1-建立具有持续广中和抗体应答的NVS特异性BMem的克隆特异性图谱，通过有限稀释分析、mAb分离和表位定位来确定不同环境抗原表位特异性BMem克隆的相对优势。目的-2-比较血浆抗体和代表BMem全克隆性的单抗之间的中和宽度，以确定重建循环抗体库的中和宽度必须汇集的单抗的数量。这些数据将被用来确定正在进行的广泛的NAB反应的克隆性。这一目标将完成对中和广度由中和特异性马赛克组成的多克隆反应而不是由一个或极少数中和特异性组成的稀有克隆反应决定的假设的检验。目前还没有针对艾滋病的疫苗。这项申请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艾滋病毒-1感染多年。这些信息应该对制造艾滋病疫苗很有用。

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.