Mechanisms of Sleep Responses to Viral Infections

描述(申请人摘要)：疲劳、嗜睡、过度 睡眠和睡眠障碍在几乎所有传染性疾病中都出现了症状 疾病。这项提议的广泛目标是描述分子 流感病毒引起睡眠改变的机制。我们 假设病毒双链(DS)RNA是在感染细胞中产生的 反过来，它会诱导包括干扰素在内的细胞因子的上调 (干扰素)。然后细胞因子诱导生长激素释放激素(GHRH) 释放后，它通过一氧化氮(NO)改善睡眠。相当大的初步 数据支持这一假设。建议的研究中使用的模型是 A/PR/8/34-HIN1流感病毒感染小鼠。PR8引起肺炎 伴随着早起的睡眠反应。在具体目标#1中，比较 将用纯净诱导的细胞因子的时间进程的基因阵列 肺和脑中的流感病毒和dsRNA。我们预计会有类似的细胞因子 两种刺激后的侧写。在特定目标#2中，GHRH受体在 病毒引起的睡眠反应将被确定。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， 在病毒挑战之后。我们预计这一调查结果将得到证实，并 生长激素替代疗法不会改变病毒诱导的睡眠反应， 但可能会降低死亡率。在特定目标#3中，一氧化氮合酶基因敲除 小鼠将被用来研究一氧化氮在病毒诱导的睡眠中的作用。 初步数据显示，在主机挑战后，睡眠反应减弱 NOS-2(诱导型一氧化氮合酶)基因敲除小鼠。在特定目标#4中，干扰素受体(I型 和ii)基因敲除小鼠将被用来研究IFN在 病毒诱导的睡眠。在具体目标2、3和4中，我们预计 病毒在突变株中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 在这些突变的小鼠品系中。在具体目标5中，我们将调查，在 体外，病毒相关dsRNA在流感诱导细胞因子中的作用。 因为我们假设dsRNA通过核因子kappa上调细胞因子 B(NFKB)我们将确定NFKB的其他激活剂，例如自由基， 对小鼠巨噬细胞NFKB激活的影响及药物阻滞剂的作用 影响病毒诱导的NFKB激活和细胞因子级联反应。这个 预期的结果将极大地帮助我们理解分子 病毒诱导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.