The evolutionary landscape of HIV broadly neutralizing antibody development

项目摘要 人体免疫机能丧失病毒（艾滋病毒）大流行病仍然是一个全球性的健康威胁， 估计每年有3 500万例活动性感染和100万例死亡。小分子药物已经改变了 艾滋病毒的预后和预防，但预防性疫苗，eliminating保护性中和抗体是一个 关键优先事项。由于艾滋病毒在全球范围内的巨大多样性，通过免疫系统诱导抗体仍然是一个开放的挑战。 疫苗接种具有足够的抗病毒广度和效力，以保护免受各种病毒分离株的侵害。广泛罕见 中和抗体（bnAbs），可以抑制大多数流行的HIV毒株已被分离， 艾滋病毒感染者，建立一个有效的艾滋病毒疫苗的目标终点。这些bnAb也是 用于直接治疗或预防用途的有前景的分子。然而，我们对HIV bnAb的理解 然而，功能和发展尚未证明足以有效利用其保护潜力。在这 建议，我概述了创新的高通量生物化学方法来研究分子进化 bnAb功能的基础。重点关注针对HIV包膜CD4受体结合位点的bnAb，我们将 使用高通量突变扫描方法来揭示选择bnAb中的所有氨基酸突变， 它们的幼稚前体影响与多种HIV Env抗原结合的特异性与广度， 设计的候选疫苗免疫原。与此同时，我们将测量突变对正交 可以限制抗体发展的抗体性质，包括热力学稳定性，多特异性， 和自身反应性。通过将这些丰富的突变数据与现有的天然抗体合成在一起， 从感染和疫苗接种的测序数据，我们将确定分子进化的力量，指导 并限制bnAb亲和力成熟。这些观察结果将使设计改进的抗体 治疗剂和疫苗免疫原，这类有前途的艾滋病毒bnAb。更广泛地说， 研究将为抗体开发的分子和进化基础提供重要的视角 并开拓跨学科的方法，用于各种病原体， 广泛的免疫识别对治疗控制提出了挑战。

PROJECT SUMMARY The human immunodeficiency virus (HIV) pandemic remains a global health threat, responsible for an estimated 35 million active infections and one million deaths per year. Small molecule drugs have transformed HIV prognosis and prophylaxis, but a preventative vaccine that elicits protective neutralizing antibodies is a critical priority. Because of HIV’s vast global diversity, it remains an open challenge to elicit antibodies via vaccination with sufficient antiviral breadth and potency to protect against diverse viral isolates. Rare broadly neutralizing antibodies (bnAbs) that can inhibit the majority of circulating HIV strains have been isolated from HIV-infected patients, establishing a targeted endpoint for an effective HIV vaccine. These bnAbs are also promising molecules for direct therapeutic or prophylactic use. However, our understanding of HIV bnAb function and development has not yet proven sufficient to effectively harness their protective potential. In this proposal, I outline innovative high-throughput biochemical approaches to study the molecular evolutionary basis for bnAb function. With a focus on bnAbs targeting the HIV Envelope CD4 receptor-binding site, we will use high-throughput mutational scanning methods to reveal how all amino acid mutations in select bnAbs and their naive precursors impact specificity versus breadth of binding to diverse HIV Env antigens and to rationally designed candidate vaccine immunogens. In parallel, we will measure mutational effects on orthogonal antibody properties that can constrain antibody development including thermodynamic stability, polyspecificity, and autoreactivity. By synthesizing this wealth of mutational data together with existing natural antibody sequencing data from infection and vaccination, we will identify the molecular evolutionary forces that guide and constrain bnAb affinity maturation. These observations will enable the design of improved antibody therapeutics and vaccine immunogens for this promising class of HIV bnAb. More broadly, the proposed studies will provide important perspectives on the molecular and evolutionary basis for antibody development and pioneer interdisciplinary methodological approaches with utility for various pathogens where the need for broad immune recognition presents a challenge to therapeutic control.