Development of a novel BSL2 system for high-throughput analysis of hantavirus entry glycoproteins

在美洲和欧亚大陆，汉坦病毒分别是汉坦病毒心肺综合征（HCPS）和肾综合征出血热（HFRS）的病原体。这些病毒通常从啮齿动物宿主传播给人类。然而，南美安第斯汉坦病毒的多起人际传播事件引起了对这些致命病毒的重大公共卫生关注，这些病毒的致死率高达40%。目前还没有fda批准的汉坦病毒疫苗和治疗方法。汉坦病毒通过其病毒粒子表面的Gn/Gc糖蛋白介导细胞进入和感染，这也是保护性免疫反应的主要靶点。然而，我们对汉坦病毒进入和抗原性中Gn/Gc的分子决定因素的理解仍然有限，至少部分原因是由于汉坦病毒研究的生物安全级别3 （BSL3）遏制的一般要求和缺乏反向遗传系统。为了解决这些限制，已经开发了多个BSL2假病毒系统。然而，目前的BSL2系统由于其低效的基于质粒的拯救而受到可扩展性差和产生的病毒多样性有限的限制，这使得它们无法更全面地分析进入糖蛋白的生物学和功能。在这里，我们建议通过开发一种新的BSL2系统来解决汉坦病毒领域的这一主要缺陷，该系统可以对Gn/Gc在病毒进入和抗原性中的作用进行全面的反向和正向遗传分析。

Hantaviruses are the causative agents of hantavirus cardiopulmonary syndrome (HCPS) and hemorrhagic fever with renal syndrome (HFRS) in Americas and Eurasia, respectively. These viruses are usually transmitted from their rodent reservoirs to human. However, multiple incidents of person-to-person transmission of a South American Andes hantavirus raises significant public health concern about these deadly viruses with up to 40% case fatality rates. No FDA-approved hantavirus vaccines and therapies exist. Cellular entry and infection of hantaviruses is mediated by its virion surface Gn/Gc glycoproteins, which are also the main target of protective immune responses. However, our understanding of the molecular determinants of Gn/Gc in hantavirus entry and antigenicity remains limited, at least partly, due to the general requirement of Biosafety level-3 (BSL3) containment for hantavirus research and the lack of a reverse genetics system. To address these limitations, multiple BSL2 pseudovirus systems have been developed. However, current BSL2 systems are limited by poor scalability and limited diversity of the generated viruses due to their inefficient plasmid-based rescue, which makes them incompatible with a more comprehensive analysis of the biology and function of the entry glycoproteins. Here, we propose to fix this major shortcoming of the hantavirus field by developing a novel BSL2 system that allows comprehensive reverse as well as forward genetic analysis of Gn/Gc’s role in virus entry and antigenicity.