SHINING A LIGHT ON BAT CELLULAR IMMUNITY FOLLOWING VIRUS INFECTION

项目摘要/摘要 蝙蝠是多种病毒的重要宿主，其中几种与死亡率有关，如 在确诊的人类病例中，高达90%。这包括高致病性的埃尼帕病毒，其中 亨德拉病毒(HEV)出现在澳大利亚，尼帕病毒(NIV)通过马和猪出现在东南亚 中间宿主分别为。埃尼帕病毒已被证明是从翼手蝙蝠传播的， 澳大利亚黑狐(Pteropus Alecto)被确认为HEV(1，2)的宿主。与其他病毒一样 蝙蝠的感染，自然或实验感染HEV的蝙蝠不会引起疾病的临床症状，尽管 病毒的脱落。P.Alecto的抗病毒免疫反应是所有蝙蝠物种中研究最充分的之一， 已经发现了新的免疫机制，包括干扰素α的结构性表达 被我们的团队发现(3)。这些特点使P.Alecto-HEV模型特别适合于 回答我们在这个项目中提出的问题。尽管越来越多的人畜共患病毒从 蝙蝠，蝙蝠免疫学的研究仍处于初级阶段，很少有研究考察适应性免疫 任何蝙蝠物种的反应。了解蝙蝠的抗病毒反应是至关重要的，如果我们要预测和 防止病毒从蝙蝠蔓延到其他敏感物种，了解其他物种的疾病发病机制 并发现新的疗法和疫苗来治疗人类和其他动物的这些疾病。 在这项研究中，我们将描述实验感染蝙蝠的先天性和获得性免疫反应 HEV以获得关于蝙蝠如何控制病毒感染的详细见解。蝙蝠的细胞免疫应答 将使用功能分析来确定在活动性感染期间激活的细胞亚群 并探索全球基因和蛋白质的表达，以表征先天和获得性免疫反应 受感染的蝙蝠。使用创新的方法在受感染的蝙蝠中识别MHC结合的HEV多肽，建立 在以前的BAT免疫表现学研究中，将为在 感染。很少有研究全面研究蝙蝠在感染过程中的免疫反应，也没有研究 已经检测了细胞介导的免疫反应的功能激活。与感染者的比较 雪貂将允许我们直接比较造成无害(蝙蝠)和致命的机制。 (雪貂)HEV感染。预期结果包括了解抗病毒反应的基本生物学 蝙蝠以及监测蝙蝠对HEV和相关病毒的免疫力的新工具的开发。 澳大利亚疾病预防中心和莫纳什大学是唯一适合进行 在这项提案中概述的工作，以及在蝙蝠免疫学方面合作的良好记录。这支队伍有 获得高度隔离的设施和专业知识，以实施危险病原体的动物感染 再加上获得蛋白质化学设施和生产四聚体试剂的专业知识。

Project summary/abstract Bats are important reservoir hosts for a variety of viruses, several of which are associated with fatality rates as high as 90% among diagnosed human cases. This includes the highly pathogenic henipaviruses, of which Hendra virus (HeV) emerged in Australia and Nipah virus (NiV) in South-east Asia via horse and pig intermediate hosts respectively. The henipaviruses have been shown to be transmitted from Pteropus bats, with the Australian black flying fox (Pteropus alecto) confirmed as a reservoir for HeV (1, 2). As with other viral infections in bats, natural or experimental infection of bats with HeV causes no clinical signs of disease despite shedding of virus. The antiviral immune response of P. alecto is among the most well studied of all bat species, with novel immune mechanisms already discovered including the constitutive expression of interferon alpha discovered by our team (3). These characteristics make the P. alecto – HeV model uniquely suited to answering the questions we propose in this project. Despite the increasing emergence of zoonotic viruses from bats, studies of bat immunology remain in their infancy and few studies have examined the adaptive immune responses of any bat species. Understanding the antiviral responses in bats is crucial if we are to predict and prevent virus spillover from bats to other susceptible species, understand disease pathogenesis in other mammals and uncover new therapeutics and vaccines to treat these diseases in humans and other animals. In this study, we will characterise the innate and adaptive immune response of experimentally infected bats to HeV to obtain detailed insights into how bats control viral infection. The cell mediated immune response of bats will be dissected using functional assays to determine the subsets of cells activated during an active infection and explore global gene and protein expression to characterise the innate and adaptive immune response of infected bats. The use of innovative approaches to identify MHC bound HeV peptides in infected bats, building on previous bat immunopeptidomics studies, will provide new insights into peptide presentation during infection. Few studies have comprehensively studied the immune response of bats during infection, and none have examined the functional activation of the cell mediated immune response. Comparison with infected ferrets will allow us to directly compare mechanisms responsible for innocuous (bats) compared to fatal (ferrets) HeV infection. Expected outcomes include understanding the basic biology of antiviral responses in bats and the development of new tools to monitor bat immunity to HeV and related viruses. The Australian Centre for Disease Preparedness and Monash University are uniquely suited for performing the work outlined in this proposal with a strong track record of working together on bat immunology. The team has access to high containment facilities and expertise to perform animal infections with dangerous pathogens combined with access to protein chemistry facilities and expertise in generating tetramer reagents.