SHINING A LIGHT ON BAT CELLULAR IMMUNITY FOLLOWING VIRUS INFECTION

项目概要/摘要 蝙蝠是多种病毒的重要储存宿主，其中一些与死亡率有关， 在确诊的人类病例中高达90%。这包括高致病性亨尼帕病毒，其中 亨德拉病毒（Hendra virus，HeV）在澳大利亚出现，尼帕病毒（Nipah virus，NiV）在东南亚通过马和猪出现 中间宿主。亨尼帕病毒已被证明是由狐蝠传播的， 澳大利亚黑狐蝠（Pteropus alecto）被确认为HeV的贮存者（1，2）。与其他病毒一样， 蝙蝠的感染，蝙蝠与HeV的自然或实验感染不会引起疾病的临床症状， 病毒脱落。蝙蝠的抗病毒免疫反应是所有蝙蝠物种中研究最充分的， 已经发现了新的免疫机制， 我们的团队（3）这些特征使得P. alecto-HeV模型独特地适合于 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病毒出现， 蝙蝠，蝙蝠免疫学的研究仍然处于起步阶段，很少有研究已经检查了适应性免疫 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， 防止病毒从蝙蝠传播到其他易感物种，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，我们将研究实验感染蝙蝠的先天性和适应性免疫反应， HeV获得蝙蝠如何控制病毒感染的详细见解。蝙蝠的细胞免疫应答 将使用功能测定法进行解剖，以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表达，以确定先天性和适应性免疫反应， 被感染的蝙蝠使用创新的方法来识别感染蝙蝠中MHC结合的HeV肽， 在以前的蝙蝠免疫肽组学研究，将提供新的见解肽呈递过程中， 感染很少有研究全面研究蝙蝠在感染过程中的免疫反应， 已经检查了细胞介导的免疫应答的功能激活。与感染者比较 雪貂将使我们能够直接比较机制负责无害（蝙蝠）相比，致命的 （雪貂）HeV感染。预期的结果包括了解抗病毒反应的基本生物学， 蝙蝠和开发新的工具来监测蝙蝠对HeV和相关病毒的免疫。 澳大利亚疾病预防中心和莫纳什大学是唯一适合执行 本建议中概述的工作，在蝙蝠免疫学方面合作的良好记录。该团队已经 获得高遏制设施和专业知识，以执行危险病原体的动物感染 结合使用蛋白质化学设施和产生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.