Extra-pulmonary pathology caused by avian H5N1 and recombinant H1N1-1918 influenz

描述（由申请方提供）：禽流感在人类中的持续暴发强调了加强研究工作以充分表征感染期间发病机制的必要性。患者的临床病程和尸检病理学表明，在呼吸道外的组织中存在流感病毒，继发于呼吸道感染但在组织中保持无病毒的疾病，或两者兼而有之。动物研究还表明，禽流感病毒的肺外传播在一定程度上和器官受累似乎取决于物种。评估非呼吸器官病理对细胞因子过度产生的确切贡献，这是人类疾病导致多器官衰竭的常见特征，对于成功的临床管理至关重要，更好地了解导致脑病发展的因素也很常见。不幸的是，感染后住院的时间，英雄的治疗努力的混杂效应，以及排除在已经严重受损的个体中进行主要器官活检的伦理考虑，需要使用动物模型来研究肺部和其他可能受感染影响的主要器官的早期感染。在过去的3年中，我们成功地优化了用于基因组应用的人流感感染的猕猴模型。为此，我们使用了一种轻度致病性的人流感病毒（A/Texas/36/91），以仔细表征临床过程，病理学和转录调控响应病毒mRNA，病毒蛋白，或活跃的病毒复制。接下来，我们将用含有来自1918年大流行流感（A/Brevig使命/1/18）的多达三个基因的德克萨斯流感病毒（据信源自禽病毒）和高致病性禽流感（A/A/Vietnam/1203/04）感染食蟹猴。除了前几年进行的工作外，我们建议通过使用系统性显微镜检查、流感抗原染色、原位免疫细胞表型分析和炎症和免疫过程标志物的免疫组织化学染色，对呼吸道以外的组织中的任何病理进行深入表征，包括环氧合酶级联反应、巨噬细胞产生一氧化氮、缺氧反应元件的诱导、血管生成等。这种方法将通过激光捕获显微切割病变，然后进行相应mRNA的定量逆转录PCR来补充和证实。我们认为，结合多年的直接经验，这种模式和使用相关的流感病毒，已知或可能是高毒力的人类，提供了一个独特的机会，阐明这一重要方面的禽流感感染的人。叙述性禽流感在人类中的爆发已经引起越来越高的发病率和死亡率。该病毒进一步适应人类宿主可能导致世界范围的流行病，如1918年或“西班牙”流感，据信是禽类来源的，并在全世界造成2000万至4000万人死亡。禽流感感染引起的人类疾病不仅限于呼吸道，而且似乎影响其他器官，导致受感染个体预后不良。我们建议在非人灵长类动物中进行深入的病理学研究，这些灵长类动物将感染高致病性禽流感病毒和与1918年流感基因重组的人类病毒，以充分表征感染对呼吸道外组织的影响，并以有助于减少感染后人类伤亡的知识体系为目标。

DESCRIPTION (provided by applicant): Ongoing outbreaks of avian influenza in humans stress the need for intensified research efforts to fully characterize the pathogenesis during infection. Clinical course in patients and post-mortem pathology have suggested either presence of influenza virus in tissues outside of the respiratory tract, disease secondary to respiratory infection but in tissues that remain virus-free, or both. Animal studies have also demonstrated extra- pulmonary spread of avian influenza viruses, to an extent and with organ-involvement that appear species- dependent. Assessing the exact contribution of non-respiratory organ pathology to cytokine overproduction, a common feature of the disease in humans leading to multi-organ failure, is critical to successful clinical management, as is a better understanding of the factors leading to development of encephalopathy, also commonly seen. Unfortunately, timing of hospitalization after infection, confounding effects of heroic therapeutic efforts, and ethical considerations precluding biopsy of major organs in already severely compromised individuals, necessitate the use of animal models to study early infection in lungs and other major organs likely affected by the infection. Over the past 3 years, we have successfully optimized a macaque model of human influenza infection for genomic applications. For this purpose, we have used a mildly pathogenic human influenza virus (A/Texas/36/91) to carefully characterize the clinical course, pathology, and transcriptional regulation in response to viral mRNA, viral proteins, or to active viral replication. Next, we will infect Cynomolgus macaques with Texas influenza virus containing up to three genes from the 1918 pandemic flu (A/Brevig Mission/1/18), believed to have originated from an avian virus, and with highly pathogenic avian influenza (A/A/Vietnam/1203/04). In addition to the work performed in previous years, we propose to do an in-depth characterization of any pathology in tissues other than the respiratory tract, through the use of systematic microscopic examination, influenza antigen staining, phenotyping of immune cells in situ, and immunohistochemistry staining for markers of inflammatory and immune processes, including the cyclooxygenase cascade, production of nitric oxide by macrophages, induction of hypoxia response elements, angiogenesis, and others. This approach will be supplemented and corroborated by laser capture microdissection of lesions followed by quantitative reverse transcription PCR for corresponding mRNAs. We believe that the combination of several years of direct experience with this model and the use of relevant influenza viruses, known to be or likely to be of high virulence in humans, provides a unique opportunity to elucidate this important aspect of avian influenza infection in humans. Bielefeldt Ohmann - R03 NARRATIVE Outbreaks of avian influenza in humans have caused increasingly high morbidity and mortality. Further adaptation of the virus to the human host is likely to result in a world-wide epidemic such as the 1918 or "Spanish" influenza, believed to have been of avian origin and to have caused 20 to 40 millions deaths worldwide. Human disease caused by infection with avian influenza is not limited to the respiratory tract, but appears to affect other organs in ways that result in poor prognosis in infected individuals. We propose to perform in-depth pathology work in non-human primates that are to be infected with a highly pathogenic avian virus and with a human virus recombined with genes from the 1918 influenza, in order to fully characterize the effect of infection on tissues outside of the respiratory tract and with the goal of contributing to the body of knowledge that will help minimize human casualties after infection.