Identification and characterization of individuals with elite anti-HIV-1 ADCC

项目摘要 广谱中和抗体(BNAbs)的分离是HIV-1抗体向前迈出的一大步 有针对性的疫苗努力。BNAbs的发现是基于两个主要的进展。第一, 高通量中和试验的建立可用于识别HIV-1感染者 它具有广泛而有力的中和反应，被称为“精英中和剂”。第二，个体的隔离 抗体产生细胞和随后的单细胞聚合酶链式反应技术使克隆和生产成为可能 高效地从精英中和者那里获得bNAbs。目前，还没有疫苗免疫原被 设计成可以在未感染的高危人群中诱导bNAb。到目前为止，几乎所有的bNAb都有 广泛的体细胞突变水平，这可能是一个特别难引发的特征 目前的疫苗技术。人类疫苗试验、相关性分析和动物研究都表明 抗体依赖性细胞毒性(ADCC)以及中和反应也是重要的 防止病毒感染。尽管ADCC的应对措施很重要，但与导致 目前还没有发现用于ADCC诱导抗体的bNAbs。例如，以前的 调查还没有发现拥有特别广泛和强大的抗HIV-1 adcc的个人 回答，被称为“精英ADCC。”从符合以下条件的个体中鉴定和随后分离抗体 具有优秀的ADCC活性可能会产生新的抗HIV-1抗体。这些诱导ADCC的抗体可能是 更易于用现有的免疫原进行诱导。具有广泛和强大的ADCC反应的个人 主要是因为化验和试剂的限制而没有被确认。我们开发了一部小说 ADCC试验将首次允许高通量筛选大量样本 ADCC的广度和效力。我们建议检查ADCC对各种病毒的反应 在治疗大量慢性感染的天真患者中。我们的研究将开始确定 拥有精英ADCC的个人。此外，我们将检查特定的抗体特征，如 与感染细胞的结合、抗体糖基化和免疫球蛋白亚型，赋予ADCC广度和 威力。未来的研究将致力于分离和鉴定从被证明患有 广泛而有力的ADCC反应。这些拟议的研究是朝着确定以下目标的第一步 新型抗HIV-1抗体，既能潜在地防止病毒感染，又能用电流诱导 疫苗免疫原。

Project Summary Isolation of broadly neutralizing antibodies (bnAbs) has been a monumental step forward in HIV-1 antibody directed vaccine efforts. The discovery of bnAbs was predicated on two major advancements. First, development of high throughput neutralization assays allowed for the identification of HIV-1 infected individuals that had broad and potent neutralization responses, termed “elite neutralizers.” Second, isolation of individual antibody producing cells and subsequent single cell PCR technologies made it possible to clone and produce bnAbs in an efficient manner from the elite neutralizers. At this time, no vaccine immunogens have been designed that can elicit bnAbs in uninfected at risk individuals. Nearly all bnAbs, isolated to date, have extensive level of somatic mutation, which may be one characteristic that will be especially difficult to elicit with current vaccine technologies. Human vaccine trials, correlate analysis, and animal studies all suggest that antibody dependent cellular cytotoxicity (ADCC) along with neutralization responses are also important in preventing virus acquisition. Despite the importance of ADCC responses, efforts similar to those that led to the discovery of bnAbs have not been undertaken for ADCC inducing antibodies. For instance, previous investigations have not identified individuals that possess exceptionally broad and potent anti-HIV-1 ADCC responses, termed “elite ADCC.” Identification and subsequent isolation of antibodies from individuals that have elite ADCC activity may yield novel anti-HIV-1 antibodies. These ADCC inducing antibodies may be more amenable for elicitation with current immunogens. Individuals with broad and potent ADCC responses have not been identified primarily because of assays and reagent limitations. We have developed a novel ADCC assay which, for the first time, will allow high throughput screening of a large number of samples for ADCC breadth and potency. We propose to examine ADCC responses against a diverse panel of viruses among a large number of chronically infected treatment naïve patients. Our studies will begin to identify individuals that harbor elite ADCC. Furthermore, we will examine if specific antibody characteristics, such as binding to infected cells, antibody glycosylation, and immunoglobulin subtype, confer ADCC breadth and potency. Future investigations will aim to isolate and characterize antibodies from individuals shown to have broad and potent ADCC responses. These proposed studies are the first step towards the goal of identifying novel anti-HIV-1 antibodies that can potentially both prevent virus acquisition and be elicited with current vaccine immunogens.