VIRAL INFECTIONS IN EPITHELIAL FUNCTION

慢性炎症、肥大和鳞状化生的正常， 鼻窦内的假性复层柱状上皮是一致的 慢性鼻窦炎的特点，是这一点的中心前提 上皮细胞功能改变可以 对这种疾病的病理生理学有贡献。这个项目将 检验一种假设，即上皮细胞的病毒感染对 在慢性鼻窦炎的发生或加重中的重要作用 引发上皮细胞功能的改变，特别是增加 产生促炎细胞因子。 为了研究病毒感染和鼻窦炎之间的联系，我们将 前瞻性监测接受过鼻窦手术的受试者。我们会 收集受试者的鼻腔和鼻窦冲洗液 疾病处于静止状态，并在他们报告后尽快 他们的鼻窦炎加重。我们将同时使用PCR和标准 一系列常见呼吸道病毒的培养方法以确定 这些样本中病毒感染的发生率，并与 感染的存在与疾病的恶化和改变 鼻部电位差(项目3)。 我们的初步数据清楚地表明，鼻病毒感染上皮细胞 体外培养的细胞会增加细胞因子的释放。治疗 含有糖皮质激素的细胞可以抑制病毒诱导的细胞因子 制作。我们将使用培养的细胞来确定分子 病毒感染和糖皮质激素调节机制的研究进展 上皮细胞细胞因子的产生。我们将使用Northern杂交分析， 核奔跑实验和导入启动子的细胞构建 确定细胞因子的产生是否在转录或转录水平上受到调节 转录后水平。核因子-kappaB和AP-1在转录水平上的作用 调控上皮细胞因子产生的调控部位 病毒和糖皮质激素也将接受检查。 为了确定上皮细胞细胞因子产生的这些变化是否可能 发生，并在鼻窦炎的恶化中起作用，在体内，我们会 慢性阻塞性肺疾病上皮细胞因子IL-8、IL-6、GM-CSF水平监测 从我们的患者群体中获得鼻腔和鼻窦冲洗 当他们的疾病处于静止期和病情恶化时。在……里面 此外，我们将使用免疫细胞化学和原位杂交来 检查一组受试者的鼻窦活检以确认 这些细胞因子的细胞来源。我们还将确定患者是否 接受糖皮质激素治疗的患者病情恶化的发生率较低 在病毒感染期间，并将任何这样的差异与 活体上皮细胞因子的产生及其与易感因素的关系 人群，如特应性状态、免疫能力或哮喘状态。 这些研究应该会为研究糖尿病的发病机制提供重要的见解。 鼻窦炎，并可能导致改进的治疗方法的发展 这一重大健康问题的治疗。我们对病毒改变的研究 上皮细胞功能的研究也可能具有更广泛的意义 与哮喘和普通感冒发病机制的相关性。

Chronic inflammation, hypertrophy and squamous metaplasia of the normal, pseudostratified columnar epithelium within the sinuses is a consistent feature of chronic sinusitis, and it is the central premise of this program application that alterations in epithelial cell function may contribute to the pathophysiology of this disease. This project will examine the hypothesis that viral infection of the epithelium plays an important role in the development or exacerbation of chronic sinusitis by triggering alterations in epithelial cell function, particularly increased production of proinflammatory cytokines. To examine the link between viral infection and sinusitis, we will prospectively monitor subjects who have undergone sinus surgery. We will collect nasal and sinus lavages from subjects during periods when their disease is quiescent and as soon as possible after they report exacerbations of their sinusitis. We will use both PCR and standard culture methods for a range of common respiratory viruses to determine the incidence of viral infection in these samples and to correlate the presence of infections with disease exacerbations and with alterations in nasal potential difference (Project 3). Our preliminary data clearly show that rhinovirus infection of epithelial cells in vitro leads to increased release of cytokines. Treatment of cells with glucocorticoids can inhibit virally-induced cytokine production. We will use cultured cells to determine the molecular mechanisms by which viral infection and glucocorticoids regulate epithelial cell cytokine production. We will use Northern blot analysis, nuclear run-on experiments, and cells transfected with promoter constructs to determine if cytokine production is regulated at the transcriptional or posttranscriptional level. The role of NF-kappaB and AP-1 transcriptional regulation sites in the control of epithelial cytokine production by viruses and glucocorticoids will also be examined. To determine if these changes in epithelial cell cytokine production may occur, and play a role in exacerbations of sinusitis, in vivo, we will monitor levels of the epithelial cytokines, IL-8, IL-6 and GM-CSF, in nasal and sinus lavages obtained from our patient populations during periods when their disease is quiescent and during exacerbations. In addition, we will use immunocytochemistry and in situ hybridization to examine sinus biopsies from a subpopulation of subjects to confirm the cellular origin of these cytokines. We will also determine if patients receiving glucocortocoids show a lower incidence of disease exacerbations during viral infections, and will relate any such differences to epithelial cytokine production in vivo and to predisposing factors in our population, such as atopic status, immune competence, or asthmatic status. These studies should provide important insights into the pathogenesis of sinusitis and may lead to the development of improved therapies for the treatment of this major health problem. Our studies on viral alterations of epithelial cell function may also have broader implications of relevance to the pathogenesis of asthma and the common cold.