Longitudinal analysis of antibody responses to HIV-1:mapping function to genotype

描述（由申请人提供）：迄今为止，还没有针对人类免疫缺陷病毒（HIV）的疫苗接种策略获得广泛成功。HIV已经发展出通过高突变率逃避体液反应的机制。在HIV-1感染者的一个子集中，已经产生了广泛中和抗体（bNAb）。这些有效的抗体靶向跨多个进化枝的病毒包膜上的保留表位。可行的疫苗接种策略可能需要确定bNAb在暴露于免疫原后进化的机制。本提案解决了以下两个问题：1）HIV-1感染者体内的抗体库如何随时间演变？2)对HIV-1的功能性抗体反应与进化阶段有何关联？这些问题将在目标1）中解决，目标1）提出对来自HIV-1感染个体的纵向B细胞库进行深度测序，使用这些序列组装系统发育树，描述具有广泛中和活性与弱中和活性的个体中抗体库的不同进化。目的2）提出对来自在目的1中深度测序的相同库的单个B细胞进行功能性抗体表型筛选。将功能表型信息与系统发育图相关联，以鉴定bNAb及其克隆祖先。这将为bNAb如何自然进化的机制提供深入了解。我们的研究结果将有助于HIV-1疫苗接种策略的发展，提供了一个蓝图的启发bNAbs。

DESCRIPTION (provided by applicant): To date no vaccination strategy against Human Immunodeficiency Virus (HIV) has been widely successful. HIV has developed mechanisms to escape the humoral response through a high mutation rate. In a subset of HIV-1 infected individuals, broadly neutralizing antibodies (bNAbs) have developed. These potent antibodies target preserved epitopes on the viral envelope across multiple clades. A viable vaccination strategy likely necessitates determining the mechanisms by which bNAbs evolve upon exposure to immunogens. The following two questions are addressed in this proposal: 1) How does the antibody repertoire evolve in HIV-1 infected individuals over time? 2) How does the functional antibody response to HIV-1 correlate to evolutionary stage? These questions will be addressed in Aim 1) which proposes to deep sequence longitudinal B-cell repertoires from HIV-1 infected individuals, using the sequences to assemble a phylogenetic tree depicting the divergent evolution of the antibody repertoires in individuals with broadly versus poorly neutralizing activity. Aim 2) proposes to perform a functional antibody phenotype screen of single B-cells from the same repertoires deep sequenced in Aim 1. The functional phenotype information will be linked to the phylogenetic map to identify bNAbs and their clonal ancestors. This will provide insight into the mechanisms of how bNAbs naturally evolve. Our results will contribute to the development of HIV-1 vaccination strategies by providing a blueprint for the elicitation of bNAbs.