Broadly Neutralizing Monoclonal Antibodies Against HIV-1

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。这一目标将在当地HIV-1感染者队列中实现，他们在没有抗逆转录病毒治疗的情况下控制自己的感染(JAIDS，50：403-8,2009)。这些人中大约有13%有广泛的中和抗体，这为我们的研究提供了研究人群。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由这些细胞表达的抗体的全长分子克隆(PNAS，106：3952-7,2009)。这种方法允许使用细胞表面表达的天然低聚HIV-1包膜糖蛋白(Env)和通过直接中和伪病毒来鉴定克隆。环状病毒反应性克隆用于产生单抗，将根据表位特异性和中和广度进行评估。这些信息将被用来检验这样的假设，即中和广度由由一个或几个中和特异性组成的单克隆或少克隆反应决定，而不是由一组中和特异性组成的多克隆反应决定。有两个具体目标。目的1-建立具有广泛中和抗体应答的NVS中Env特异性BMem的克隆性特异谱，通过有限稀释分析、mAb分离和表位定位来确定抗Env应答的克隆性特异谱，以鉴定广中和单抗。目的-2-比较代表BMem全克隆谱的血浆抗体和单抗之间的中和宽度，以确定重建循环抗体库的中和谱必须汇集的单抗的数量。单抗的中和广度将使用标准化的假病毒检测来确定。这些数据将被用来确定正在进行的广泛中和抗体反应的克隆性。这一目标将检验一种假设，即中和广度是由由一个或几个中和特异性组成的单克隆或少克隆反应决定的，而不是由一组中和特异性组成的多克隆反应决定的。 公共卫生相关性：这项研究与公共卫生相关，因为它试图了解感染艾滋病病毒的人如何做出保护性免疫反应。这些信息将有助于指导艾滋病病毒疫苗的开发。

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 local cohorts of HIV-1 infected individuals who control their infections in the absence of anti-retroviral therapy (JAIDS, 50:403-8, 2009). Approximately 13% of these individuals have circulating broadly neutralizing antibodies providing the study population for our studies. 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 molecular clones of the antibodies expressed by these cells (PNAS, 106:3952-7, 2009). This method permits clone identification using native oligomeric HIV-1 envelope glycoproteins (Env) expressed on cell surfaces and by direct neutralization of pseudoviruses. Env-reactive clones are used to produce mAbs that will be evaluated for epitope specificity and neutralization breadth. This information will be used to test the hypothesis that neutralization breadth is determined by monoclonal or pauciclonal responses comprised of one or a very few neutralizing specificities as opposed to a polyclonal response comprised of a mosaic of neutralizing specificities. 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 identify broadly neutralizing mAbs. 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 profile of the circulating antibody pool. Neutralization breadth of mAbs will be determined using standardized pseudovirus assays. This data will be used to determine the clonality of an ongoing broadly neutralizing antibody response. This aim will test the hypothesis that neutralization breadth is determined by monoclonal or pauciclonal responses comprised of one or a very few neutralizing specificities as opposed to a polyclonal response comprised of a mosaic of neutralizing specificities. PUBLIC HEALTH RELEVANCE: This research is relevant to public health in that it seeks to understand how people who are infected with the AIDS virus make protective immune responses. This information will help guide the development of a vaccine against the AIDS virus.