Mechanisms of Sleep Responses to Viral Infections

描述（申请人摘要）：疲劳、过度嗜睡、过度 睡眠和睡眠障碍几乎在所有感染性疾病中都有症状。 疾病该提案的主要目标是表征分子 流感病毒引起的睡眠变化的机制。我们 假设病毒双链（ds）RNA在感染细胞中产生 进而诱导细胞因子包括干扰素的上调 （IFN）。细胞因子然后诱导生长激素释放激素（GHRH） 一氧化氮（NO）的释放，并通过它，增强睡眠。实质性初步 数据支持这一假设。拟议研究中使用的模型是 A/PR/8/34-HIN 1流感病毒感染小鼠。PR 8导致肺炎 伴随着睡眠反应的早期发作。在具体目标#1中，比较 将使用基因阵列的时间进程的细胞因子诱导的纯 肺和脑中流感病毒和dsRNA。我们希望类似的细胞因子 两种刺激后的轮廓。在具体目标#2中，GHRH受体在 将确定病毒诱导的睡眠反应。初步数据显示 缺乏功能性GHRH受体的小鼠睡眠更少，而不是更多， 病毒攻击后。我们预计这一发现将得到证实， GH替代疗法不会改变病毒诱导的睡眠反应， 但可以降低死亡率。在特定目标#3中，一氧化氮合酶敲除 小鼠将被用于研究NO在病毒诱导的睡眠中的作用。 初步数据表明，在宿主攻击后， NOS-2（诱导型NOS）敲除小鼠。在具体目标#4中，IFN受体（I型 和II）敲除小鼠将用于研究IFN在以下中的作用： 病毒引起的睡眠在具体目标2、3和4中，我们预计， 在突变株中由病毒诱导的细胞因子基因谱将不同 并将反映病毒诱导的不同睡眠反应 在这些突变品系的小鼠中。在具体目标#5中，我们将研究， 体外，病毒相关dsRNA在流感诱导细胞因子中的作用。 由于我们假设dsRNA通过核因子κ上调细胞因子， B（NF κ B）我们将确定NF κ B的其他激活剂，例如，自由基， 对小鼠巨噬细胞中NF κ B活化的影响以及药理学阻断剂 影响病毒诱导的NF κ B活化和细胞因子级联反应。的 预期的结果将极大地帮助我们了解分子 病毒引起的睡眠反应和其他方面的机制， 急性期反应

DESCRIPTION (applicant's abstract): Fatigue, excessive sleepiness, excess sleep, and sleep disturbances are presenting symptoms in nearly all infectious diseases. The broad objective of this proposal is to characterize the molecular mechanisms responsible for changes in sleep induced by influenza virus. We hypothesize that viral double-stranded (ds) RNA is produced in infected cells and it, in turn, induces an upregulation of cytokines including interferons (IFN). The cytokines then induce growth hormone releasing hormone (GHRH) release and it, via nitric oxide (NO), enhances sleep. Substantial preliminary data support this hypothesis. The model used in the proposed studies is A/PR/8/34-HIN1 influenza virus infection in the mouse. PR8 causes a pneumonitis accompanied by early onset of sleep responses. In Specific Aim #1, a comparison will be made using gene arrays of the time courses of cytokines induced by pure influenza virus and dsRNA in lung and brain. We expect a similar cytokine profile after both stimuli. In Specific Aim #2 the role of the GHRH receptor in viral-induced sleep responses will be determined. Preliminary data indicate that mice lacking a functional GHRH receptor sleep less, rather than more, after viral challenge. We anticipate that that finding will be confirmed and that GH replacement therapy will not alter the virus-induced sleep responses, but may reduce mortality. In Specific Aim #3, nitric oxide synthase knockout mice will be used to investigate the role of NO in viral-induced sleep. Preliminary data indicate an attenuated sleep response after host challenge in NOS-2 (inducible NOS) knockout mice. In Specific Aim #4, IFN receptor (types I and II) knockout mice will be used to investigate the role of IFNs in viral-induced sleep. In Specific Aims 2, 3, and 4 we anticipate that the cytokine gene profiles induced by virus in the mutant strain will be different from controls and will reflect the different sleep responses induced by virus in these mutant strains of mice. In Specific Aim #5, we will investigate, in vitro, the role of virus-associated dsRNA in cytokine induction by influenza. Since we hypothesize that dsRNA upregulates cytokines via nuclear factor kappa B (NFKB) we will determine what other activators of NFKB, e.g., free radicals, do to NFKB activation in murine macrophages and how pharmacological blockers affect viral-induced activation of NFKB and the cytokine cascade. The anticipated results will greatly aid our understanding of the molecular mechanisms involved in viral-induced sleep responses and other facets of the acute phase response.