Virus and Antibody Gene Sequencing Core

项目摘要 HIV-1感染者和SHIV中广泛交叉反应中和抗体（bNAb）的开发 感染的恒河猴（RM）需要反复多轮的包膜糖蛋白（Env）介导的B细胞 受体刺激、中和抗体（NAb）激发和相关病毒逃逸。在这种病毒/抗体中 “军备竞赛”，产生特定的Env逃逸突变体，在无数的其他Env变体中， 存在于感染个体中，能够引发和加强能够发展的抗体谱系 中和宽度虽然病毒/抗体协同进化的原理已经确立，但数量， 启动和维持bNAb谱系的特定Env变体的身份、继承和合作 成熟仍然是未知的。在这个更新的应用程序中，这个HIVRAD团队建议使用猿猴- 人类免疫缺陷病毒（SHIV）感染恒河猴（RM）作为HIV-1分子指南 诱发V3高甘露糖斑块bNAb的Env免疫原。与项目1和项目3合作，我们将跟踪 在RM中Env-抗体共进化的模式，其开发V3聚糖补丁bNAb，然后使用该 识别特异性Env变体的信息，所述特异性Env变体引发这些中和反应并使其亲和力成熟 宽度该小组最近的工作为这些目标提供了强有力的支持（1-6）。首先，研究小组发现， 感染各种携带HIV-1 Env的SHIV的150个RM中的24个中的异源中和宽度，包括 9只动物具有V3高甘露糖斑块bNabs（参见项目1的初步数据）。第二，Env抗体 在SHIV感染的RM中的协同进化被发现与在相应的 人类，包括类似的免疫遗传学，表位识别的结构和化学解决方案，以及近- 相同的Env逃逸途径响应于Nab压力（1-5）。最后，团队设计了一个合理的策略， 用于顺序免疫原设计，以避开通过疫苗接种诱导bNAb的困难障碍（6）。我们 因此，假设在被生殖系靶向CH 848感染的RM中Env-抗体共进化的表征 和BG505.N332 SHIV将识别Env免疫型，当构建为纳米颗粒或SOSIP Env时， 三聚体并用于免疫RM，将引发V3聚糖bNAb。病毒和抗体测序核心（核心 B）将继续通过提供最先进的病毒和抗体测序来支持这个HIVRAD团队 服务以及独特的病毒学专业知识。与生物信息学和统计学核心密切合作 (Core C）以及项目1，2和3，我们将（i）描述SHIV感染中进化的env准种 产生V3聚糖bNAb以绘制中和逃逸途径的猕猴（Aim 1），和（ii）序列 SHIV感染前后来自恒河猴记忆B细胞的抗体重链和轻链V（D）J可变区 以及免疫后的敲入小鼠，以促进bNAb谱系追踪并确定抗原的作用 B细胞命运中的受体亲和力（Aim 2）。目标是确定Env-抗体共进化所需的关键步骤， V3聚糖bNAb的开发，可以快速转化为合理的免疫原设计。

PROJECT SUMMARY The development of broadly cross-reactive neutralizing antibodies (bNAbs) in HIV-1 infected humans and SHIV infected rhesus macaques (RMs) requires iterative rounds of envelope glycoprotein (Env)-mediated B cell receptor stimulation, neutralizing antibody (NAb) elicitation, and associated virus escape. In this virus/antibody “arms race”, particular Env escape mutants are generated which, among the myriads of other Env variants present in infected individuals, are able to prime and boost antibody lineages that are capable of developing neutralization breadth. Although the principle of virus/antibody co-evolution has been established, the number, identity, succession, and cooperation of the particular Env variants that initiate and sustain bNAb lineage maturation remain largely unknown. In this renewal application, this HIVRAD team proposes to use simian- human immunodeficiency virus (SHIV) infected rhesus macaques (RMs) as a molecular guide to develop HIV-1 Env immunogens that elicit V3 high mannose patch bNAbs. In collaboration with Projects 1 and 3, we will trace the patterns of Env-antibody coevolution in RMs that develop V3 glycan patch bNAbs and then use this information to identify specific Env variants that prime and affinity-mature these responses towards neutralization breadth. Recent work by the group provides strong support for these goals (1-6). First, the team identified heterologous neutralization breadth in 24 of 150 RMs infected with various HIV-1 Env bearing SHIVs, including nine animals with V3 high mannose patch bNabs (see Preliminary Data of Project 1). Second, Env-antibody coevolution in SHIV infected RMs was found to be strikingly similar to that observed in the corresponding humans, including similar immunogenetics, structural and chemical solutions to epitope recognition, and near- identical Env escape pathways in response to Nab pressures (1-5). Finally, the team devised a rational strategy for sequential immunogen design to circumvent difficult roadblocks in bNAb induction by vaccination (6). We thus hypothesize that characterization of Env-antibody coevolution in RMs infected by germline-targeted CH848 and BG505.N332 SHIVs will identify Env immunotypes that, when constructed as nanoparticles or SOSIP Env trimers and used to immunize RMs, will elicit V3 glycan bNAbs. The Virus and Antibody Sequencing Core (Core B) will continue to support this HIVRAD team by providing state-of-the-art virus and antibody sequencing services as well as unique virology expertise. Working closely with the Bioinformatics and Statistics Core (Core C) as well as Projects 1, 2 and 3, we will (i) characterize the evolving env quasispecies in SHIV infected macaques that develop V3 glycan bNAbs to map neutralization escape pathways (Aim 1), and (ii) sequence antibody heavy and light chain V(D)J variable regions from rhesus memory B cells before and after SHIV infection as well as knock-in mice after immunization to facilitate bNAb lineage tracing and define the role of antigen receptor affinity in B cell fates (Aim 2). The goal is to identify key steps in Env-antibody co-evolution required for V3 glycan bNAb development that can be rapidly translated into rational immunogen design.