Broad Neutralizing Monoclonal Antibodies From HIV Controllers

描述（由申请人提供）：本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，这些感染者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，并且具有循环广泛中和抗体（广泛nAb）。我们的方法的一个关键要素是开发一种新的测定来普查Env特异性记忆B细胞克隆（BMem），其允许快速和直接克隆全长单克隆抗体（mAb）。将对这些mAb的表位特异性和中和宽度进行表征，以创建在HIV-1感染控制期间产生的BMem的克隆谱。该信息将用于检验以下假设：中和宽度由多克隆反应（由中和特异性镶嵌组成）决定，而不是由一种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并在体外阻断感染的新型mAb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-为了开发来自具有正在进行的广泛中和抗体应答的NVS的Env特异性BMem的克隆特异性谱-将通过有限稀释分析、mAb分离和表位作图来确定抗Env应答的克隆特异性谱，以确定对不同Env表位特异性的BMem克隆的相对优势。目的-2-比较血浆抗体和代表BMem完整克隆谱的mAb之间的中和宽度，以确定必须合并的mAb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应答的克隆性。这一目的将完成对以下假设的检验，即中和宽度是由多克隆反应（由中和特异性镶嵌组成）决定的，而不是由一种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HIV-1感染多年。这些信息对研制艾滋病疫苗应该是有用的。

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.