PPG: SARS-CoV-host cell interactions and vaccine development

概览-摘要 2002- 2003年严重急性呼吸系统综合症（SARS）的出现和中东 2012年的呼吸综合征（MERS）表明，人畜共患冠状病毒（CoV）已经并将 可能继续从人畜共患病源传播到感染人群。MERS-CoV继续 在骆驼中传播并传播给易感人群，强调需要更好地了解 致病性人呼吸道CoV介导的疾病的发病机制。在这个PPG中， 具有冠状病毒发病机制、分子生物学、免疫学和 疫苗学将共同努力，以了解病毒因素和失调的先天和适应性 免疫应答导致年轻和老年动物以及动物中MERS和SARS疾病 合并症所有这些项目都将利用新开发的表达人类MERS的小鼠 受体（DPP 4）代替小鼠受体（hDPP 4-KI）和小鼠毒性MERS-CoV，选择 在这些小鼠中（MERSMA）。项目1将使用MERSMA来研究衰老在感染小鼠中的作用。 项目1也是基于已发表的数据，显示具有抗炎作用的特定eiconsanoids 在衰老过程中，蛋白质及其上游磷脂酶增加，导致免疫延迟， SARS-CoV（以及扩展后，可能是MERSMA）感染后的反应。项目2基于 初步数据显示MERS-CoV对病毒宿主细胞蛋白酶的依赖性更大 比SARS-CoV更容易进入。该项目将研究在表面（S）中发现的独特突变。 MERSMA的糖蛋白，似乎影响蛋白酶功能。项目3将研究如何 MERSMA比最初的人类EMC/2012菌株引起更严重的疾病，重点是 ORF 4 b辅助蛋白。该项目还将研究hDPP 4如何促进疾病 严重性。项目4基于已发表的数据，显示CoV E蛋白具有离子通道活性， 是一种毒力因子，含有PDZ结合结构域（PBM），这对病毒活力至关重要。这 该项目将重点关注E蛋白如何引起肺水肿以及PBM在肺水肿中的作用。 发病机制在小鼠传代过程中，MERSMA的E蛋白C端出现了一个新的PBM， 角色将被研究。该项目还将继续开发安全的MERS和SARS减毒活疫苗 疫苗。所有项目都将使用动物/病毒学核心，该核心将提供非重组和 重组MERS-CoV和SARS-CoV，并将监测和分析感染的小鼠。使用核心 为这些目的将最大限度地提高实验质量控制和有效利用我们的资源。 这些项目都是相互关联和协作的，将利用独特的技能， 提供有关MERS和SARS发病机制的新信息 这对疫苗的研发至关重要。

OVERVIEW-SUMMARY The emergence of the Severe Acute Respiratory Syndrome (SARS) in 2002-3 and the Middle East Respiratory Syndrome (MERS) in 2012 demonstrates that zoonotic coronaviruses (CoV) have and will likely continue to spread from zoonotic sources to infect human populations. MERS-CoV continues to circulate in camels and to spread to susceptible humans, highlighting the need to better understand the pathogenesis of diseases mediated by pathogenic human respiratory CoV. In this PPG, investigators with experience in coronavirus pathogenesis, molecular biology, immunology and vaccinology will work together to understand how virus factors and dysregulated innate and adaptive immune responses contribute to MERS and SARS disease in young and aged animals and in animals with co-morbidities. All of the projects will utilize newly developed mice expressing human MERS receptor (DPP4) in lieu of the mouse receptor (hDPP4-KI) and a mouse-virulent MERS-CoV, selected in these mice (MERSMA). Project 1 will use MERSMA to investigate the role of aging in infected mice. Project 1 is also based on published data showing that specific eiconsanoids with anti-inflammatory properites and their upstream phopholipases increase during aging, contributing to a delayed immune response after SARS-CoV (and by extension, perhaps MERSMA) infection. Project 2 is based on preliminary data showing that MERS-CoV has a greater dependence on host cell proteases for virus entry than does SARS-CoV. This project will investigate unique mutations found in the surface (S) glycoprotein of MERSMA that appear to affect protease function. Project 3 will investigate how MERSMA causes more severe disease than the initial human EMC/2012 strain, with focus on the ORF4b accessory protein. This project will also investigate how hDPP4 contributes to disease severity. Project 4 is based on published data showing that the CoV E protein has ion channel activity, is a virulence factor and contains a PDZ binding domain (PBM), which is critical for virus viability. This project will focus on how the E protein causes edema in lungs and on the role of the PBM in pathogenesis. A novel PBM in the C terminal of E arose in MERSMA during mouse passage and its role will be studied. This project will also continue to develop safe, live attenuated MERS and SARS vaccines. All of the projects will use the Animal/Virology Core, which will provide nonrecombinant and recombinant MERS-CoV and SARS-CoVs and will monitor and analyze infected mice. Using the Core for these purposes will maximize experimental quality control and effective use of our resources. These projects are all interrelated and collaborative, will take advantage of the unique skills and expertise of the project directors and provide new information about MERS and SARS pathogenesis that is essential to vaccine development.