Establishment of a Bat Resource for Infectious Disease Research

修改后的项目摘要/摘要部分 项目概述/摘要蝙蝠是许多人畜共患病病毒的宿主或疑似宿主，包括SARS、SARS2和MERS冠状病毒、Nipah和Hendra病毒以及埃博拉和马尔堡病毒。人们对这些病毒如何在蝙蝠储藏室中循环知之甚少，主要是因为缺乏可用于开发实验感染模型的蝙蝠菌落。为了解决这一不足，我们将在孟加拉国捕获马蹄蝠和印度飞狐，它们分别是尼帕病毒和SARS相关冠状病毒的宿主，在那里它们将被隔离，并在它们适应圈养的过程中提供兽医护理。蝙蝠将被运往密歇根州立大学建立繁殖群体，作为研究这些病毒的研究人员的资源。我们将从各种组织中产生原代细胞培养和永生化细胞系，并冷冻活的骨髓，这将有助于研究这些病毒如何感染蝙蝠细胞，以及病毒可能如何调节先天免疫反应。还将为研究界生产重组细胞因子，包括产生巨噬细胞和树突状细胞(GM-CSFFlt3L)、T细胞(IL-2)以及体内调节适应性免疫反应(干扰素γ、IL-4)的细胞因子。此外，我们还将生产用于细胞因子检测和免疫细胞亚群的流式细胞术以及体内中和的单抗。最后，我们将进行尼帕病毒、SARS-CoV-2和SARS相关冠状病毒BANAL-52和BANAL-236的实验性感染研究，以研究感染动力学、病毒分布以及蝙蝠感染期间的转录、蛋白质和代谢组学特征，以及免疫应答的升级和消退。来自幼稚和受感染蝙蝠的组织、细胞和血清将被存档在生物库中，供研究社区使用。这一资源的建立将有助于更好地了解蝙蝠如何在没有疾病的情况下携带高致病性病毒，并可能揭示增加对人类的溢出风险的事件。反过来，这些信息可能导致制定缓解战略，以防止未来的病毒溢出，并发现治疗冠状病毒和尼帕病毒疾病的新战略。

Modified Project Summary/Abstract Section Project Summary / Abstract Bats are reservoirs, or suspected reservoirs, of many zoonotic viruses, including SARS, SARS2 and MERS coronaviruses, Nipah and Hendra viruses, and Ebola and Marburg viruses. Little is known about how these viruses circulate in their bat reservoirs, principally because of a lack of bat colonies that can be used for the development of experimental infection models. To address this deficiency, we will capture horseshoe bats and Indian flying foxes, respective reservoir hosts of Nipah virus and SARS-related coronaviruses, in Bangladesh where they will be quarantined and provided veterinary care as they adapt to captivity. Bats will be shipped to CSU to establish the breeding colonies as a resource for investigators who study these viruses. We will generate primary cell cultures and immortalized cell lines from various tissues and freeze live bone marrow that will be useful for studying how these viruses infect bat cells, and how the viruses may modulate the innate immune responses. Recombinant cytokines will also be produced for the research community, including those for generating macrophages and dendritic cells (GM-CSF, Flt3L), T cells (IL-2) and for in vivo modulation of the adaptive immune response (IFNγ, IL-4). Moreover, we will generate monoclonal antibodies for use in cytokine detection assays and flow cytometry of immune cell subsets and in vivo neutralization. Finally, we will perform experimental infection studies of Nipah virus, SARS-CoV-2 and the SARS-related coronaviruses, BANAL-52 and BANAL-236, to study the infection kinetics, virus distribution and transcriptomic, proteomic and metabolomic profiles of bats during infection, and escalation and resolution of the immune response. Tissues, cells and sera from naïve and infected bats will be archived in a biobank that will be made available to the research community. The establishment of this resource will lead to a better understanding of how bats host highly pathogenic viruses without disease and may shed light on events that increase spillover risks to humans. In turn, this information could lead to development of mitigation strategies to prevent future virus spillover and uncover new strategies for therapeutic treatment of coronavirus and Nipah virus diseases.