Molecular-level mapping of the human antibody response to hantavirus infection

正如当前的全球大流行所强调的那样，能够迅速应对进入人群的新出现的致病性病毒物种，对全球卫生至关重要。快速反应在很大程度上取决于事先了解病毒编码糖蛋白表面的抗原区域，以及在感染期间如何通过中和抗体靶向它们。汉坦病毒从小型啮齿动物传播给人类后，可引起严重疾病，包括汉坦病毒肺综合征和肾综合征出血热。由于缺乏预防性疫苗和有效治疗，这一分布在全球的新出现的病毒群对人类健康构成了持续的负担。本研究旨在揭示人体免疫反应对汉坦病毒感染的反应。我们将提供分子水平的细节，如何中和抗体，这是恢复和防止感染的关键，靶向和阻碍汉坦病毒表面显示糖蛋白的功能。由于这些信息对于抗汉坦病毒对策的合理发展至关重要，因此从本研究中收集的信息将对抗病毒和疫苗设计工作至关重要，从而提高我们应对这些病原体出现的能力。在我们的项目中，我们将从以前感染过Puumala病毒的个体中分离中和抗体和交叉中和抗体（nab和crnab）。我们将采用多学科结构（x射线晶体学和低温电子断层扫描）和功能生物学方法来确定自然感染期间引发的nab和crnab如何能够预防病毒感染。该项目将解决四个基本和转化生物医学问题：1。汉坦病毒表面显示的两种糖蛋白Gn或Gc中，哪一种糖蛋白在汉坦病毒感染后更能被nab靶向？我们能否确定遗传上不同且生物医学上相关的汉坦病毒中保守的Gn或Gc上的易感性位点？本项目产生的强效汉坦病毒nab和crnab的作用机制是什么？它们在宿主细胞进入的哪个阶段阻碍宿主细胞进入？我们能否确定人类nab和crnab的协同组合可以预防汉坦病毒感染？通过了解人类抗体介导的汉坦病毒感染中和的分子和机制基础，这项研究将通过以下方式提高我们对新汉坦病毒和已建立的汉坦病毒的出现的准备：(i)对这些重要病原体的生物学提供新的见解；（ii）提供一个蓝图，将增强抗汉坦病毒生物制剂的开发，包括用于疾病治疗的协同抗体鸡尾酒和泛汉坦病毒候选疫苗。

As highlighted by the current global pandemic, an ability to respond rapidly to newly emerging pathogenic viral species that cross into the human population is essential for global health. A rapid response is greatly dependent upon prior knowledge of the antigenic regions presented on the surface of virally encoded glycoproteins and how they are targeted by neutralising antibodies during infection. Upon transmission from small rodents to humans, hantaviruses cause severe disease, including hantavirus pulmonary syndrome and haemorrhagic fever with renal syndrome. The paucity of preventative vaccines and effective treatments, renders this globally distributed group of emerging viruses a persistent burden upon human health. This proposal aims to reveal how the human immune response reacts to hantavirus infection. We will provide molecular-level detail for how neutralising antibodies, which are key for recovery and protecting against infection, target and impede the functionality of hantavirus surface-displayed glycoproteins. As this information is essential for the rational development of anti-hantavirus countermeasures, the information gleaned from this study will be essential for anti-viral and vaccine design efforts, improving our ability to respond to the emergences of these pathogens. In our project, we will isolate neutralising and cross-neutralising antibodies (nAbs and crnAbs) from individuals previously infected with Puumala virus. We will adopt a multi-disciplinary structural (X-ray crystallography and cryo-electron tomography) and functional biology approach to determine how nAbs and crnAbs elicited during natural infection are able to prevent virus infection. This project will address four basic and translational biomedical questions:1. Is one of the two glycoproteins displayed on the hantavirus surface, Gn or Gc, targeted more efficiently by nAbs following hantavirus infection?2. Can we identify sites of vulnerability on Gn or Gc that are conserved amongst genetically distinct and biomedically relevant hantaviruses?3. What are the mechanism(s) of action utilised by potent hantaviral nAbs and crnAbs generated in this project and at what stage of host-cell entry do they impede?4. Can we identify synergistic combinations of human nAbs and crnAbs that can prevent hantavirus infection?By understanding the molecular and mechanistic basis of the human antibody-mediated neutralisation to hantavirus infection, this research will improve our readiness for the emergence of new and established hantaviruses by: (i) providing new insights into the biology of these important pathogens, and (ii) providing a blueprint that will empower the development of anti-hantavirus biologics, including synergistic antibody cocktails for use in disease treatment and pan-hantavirus vaccine candidates.