Molecular Mechanisms of Sleep Responses to Viral Infection

睡眠对病毒感染反应的分子机制

• 批准号: 7802843
• 负责人: JAMES Martin KRUEGER
• 金额: $ 30.46万
• 依托单位: WASHINGTON STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-04-15 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7802843
• 关键词: Acids; Acute; Acute-Phase Reaction; Address; Affect; Age; Animals; Antibodies; Antigens; Appearance; Attention; Binding; Biological Response Modifiers; Body Temperature; Body Temperature Changes; Body Weight; Body Weight Changes; Body Weight decreased; Brain; Cells; Cessation of life; Chronic; Communicable Diseases; Cytokine Activation; Data; Detection; Development; Disease; Dose; Double-Stranded RNA; Electron Microscopy; Encephalitis; Encephalopathies; Enzymes; Epidemic; Exposure to; Fatigue; Fever; Fiber; Generic Drugs; Genetic Transcription; Glycoproteins; Goals; Grant; HIV-1; Harvest; Hospitalization; Hour; Human; Human Herpesvirus 4; Hypothalamic structure; Immune; Immunity; Immunization; Immunoelectron Microscopy; Immunohistochemistry; Infection; Infectious Mononucleosis; Inflammatory; Influenza; Influenza A Virus, H1N1 Subtype; Injection of therapeutic agent; Interferon Type I; Interferons; Interleukin-1; Interleukins; Invaded; Kinetics; Label; Laboratories; Life; Lung; Lung diseases; Macrophage Inflammatory Proteins; Maps; Measures; Mediating; Mediator of activation protein; Messenger RNA; Microinjections; Modeling; Molecular; Motor Activity; Mus; Nasal Lavage Fluid; Nasal cavity; Nerve; Nested PCR; Neural Pathways; Nose; Nucleoproteins; Olfactory Nerve; Operative Surgical Procedures; Pathway interactions; Patients; Pattern; Pneumonia; Polymerase Chain Reaction; Principal Investigator; Production; Property; Protozoan Infections; REM Sleep; RNA; Rabies virus; Recombinants; Relative (related person); Research Personnel; Research Support; Rest; Reverse Transcriptase Polymerase Chain Reaction; Rhinovirus; Role; Saline; Serum; Sleep; Small Inducible Cytokine A3; Sudden infant death syndrome; Symptoms; TLR7 gene; Techniques; Temperature; Testing; Time; Transcript; Tumor Necrosis Factor-alpha; Tumor Necrosis Factors; Up-Regulation; Viral; Viral Antigens; Viral Encephalitis; Viral Proteins; Virion; Virus; Virus Activation; Virus Diseases; Work; acquired immunity; base; cell type; chemokine; cytokine; early onset; experience; extracellular; flu; human TLR3 protein; in vivo; induced hypothermia; infancy; influenzavirus; insight; mRNA Expression; man; natural hypothermia; neutralizing antibody; non rapid eye movement; olfactory bulb; oligoadenylate; prevent; programs; receptor binding; respiratory; response; sleep abnormalities; viral RNA

DESCRIPTION (provided by applicant): Influenza is a respiratory viral infection often accompanied by severe symptoms such as high fevers and a profound need to sleep. Collectively, these symptoms are part of the acute phase response (APR). The understanding of the molecular mediators of the viral APR is in its infancy. Some of the progress made in deciphering centrally regulated APRs resulted from research supported by this grant. Our accomplishments include; showing that sleep changes over the course of an influenza virus infection; demonstrating that a toxic viral by-product, double-stranded (ds) RNA, is found in lungs of infected mice and that it induces the APR; characterizing the influenza APR in mice, and investigating APR molecular mechanisms through development of appropriate infection models and using genetically-deficient mice. Most importantly, we recently demonstrated that strains of human influenza .previously considered incapable of accessing the mouse brain, in fact do so within a few hours following intranasal infection. In brain, viral RNA and viral antigen are accompanied by induction of cytokines with the potential for mediating the APR. We expand upon these data in Aims 1 and 2. Aim 1 addresses the general hypothesis that virus, viral proteins and/or viral RNA pass from the nasal cavities to the olfactory bulb (OB) via the olfactory nerve and induce cytokine production. We will use immunohistochemistry, polymerase chain reaction techniques, and electron microscopy to determine the time course of viral invasion of the OB and its relationship to cytokine expression. Double-labeling techniques will be used to determine co-localization of virus protein with cytokines and the cell types expressing cytokines and containing virus proteins. Aim 2 tests the hypothesis that OB-expressed cytokines activate hypothalamic (HT) cytokines and subsequent HT-regulated APRs. The timing of HT cytokine expression relative to OB expression will be determined. The effects of TNFa injection into the OB and the effects of cutting OB-brain connections on HT-cytokine and sleep and fever responses will be determined. Aim 3 tests the general hypothesis that invasion of the CNS by influenza virus is affected by immune status of the host. We determine whether homologous immunity to influenza virus offers partial protection against viral invasion of the brain. The measures made in Aim 1 will be used in Aims 2 and 3 along with sleep, body temperature, locomotor activity and body weight determinations. Aim 4 is derived from our past work showing that viral dsRNA induces flu-like symptoms. Thus mice deficient in toll- like receptor 3 (TLR3), a receptor that binds dsRNA, will be challenged with virus. APRs including sleep and brain cytokine expression will be determined. There is an association between upper respiratory viral infections and sudden infant death syndrome (SIDS); SIDS occurs during sleep and is associated with sleep abnormalities. The proposed work will provide insights into how viruses induce disease symptoms as well as such complications as SIDS and viral encephalitis/encephalopathy.

描述（由申请人提供）：流感是一种呼吸道病毒感染，通常伴有严重症状，如高烧和迫切需要睡眠。总的来说，这些症状是急性期反应（APR）的一部分。对病毒APR的分子介质的了解尚处于起步阶段。在破译中央监管的apr方面取得的一些进展是由该基金支持的研究取得的。我们的成就包括：显示在流感病毒感染过程中睡眠会发生变化；证明在受感染小鼠的肺部发现了一种有毒的病毒副产物——双链RNA，并诱导了APR；通过建立合适的感染模型和使用基因缺陷小鼠来研究小鼠流感APR的分子机制。最重要的是，我们最近证明了人类流感病毒株。以前被认为无法进入小鼠大脑，实际上在鼻内感染后的几个小时内就可以进入。在大脑中，病毒RNA和病毒抗原伴随着细胞因子的诱导，具有介导apr的潜力。我们在Aims 1和Aims 2中对这些数据进行了扩展。目的1阐述了病毒、病毒蛋白和/或病毒RNA通过嗅觉神经从鼻腔传递到嗅球并诱导细胞因子产生的一般假设。我们将使用免疫组织化学、聚合酶链反应技术和电子显微镜来确定病毒侵入OB的时间过程及其与细胞因子表达的关系。双标记技术将用于确定病毒蛋白与细胞因子的共定位以及表达细胞因子和含有病毒蛋白的细胞类型。目的2验证ob表达的细胞因子激活下丘脑（HT）细胞因子和随后的HT调节的APRs的假设。将确定HT细胞因子相对于OB表达的时间。在OB中注射TNFa和切断OB-脑连接对ht细胞因子和睡眠和发热反应的影响将被确定。目的3验证流感病毒入侵中枢神经系统受宿主免疫状态影响的一般假设。我们确定对流感病毒的同源免疫是否对病毒入侵大脑提供部分保护。在Aims 1中所做的测量将与睡眠、体温、运动活动和体重测定一起用于Aims 2和3。Aim 4源于我们过去的工作，表明病毒dsRNA诱导流感样症状。因此，缺乏toll样受体3 (TLR3)（一种结合dsRNA的受体）的小鼠将受到病毒的攻击。包括睡眠和脑细胞因子表达在内的apr将被确定。上呼吸道病毒感染与婴儿猝死综合症（SIDS）之间存在关联；SIDS发生在睡眠中，与睡眠异常有关。拟议的工作将为病毒如何诱发疾病症状以及诸如小岛屿发展中国家和病毒性脑炎/脑病等并发症提供见解。