ACTIVATION OF NUCLEAR FACTOR KAPPA BETA BY CALCIUM IN AIRWAY EPITHELIAL CELLS

囊性纤维性肺病以中性粒细胞为主 呼吸道炎症，最终导致纤维化，支气管扩张， 和肺功能衰竭。铜绿假单胞菌感染是一种重要的 这一炎症过程的原因。在提出的实验中，准确地说 这种微生物如何刺激正常和上皮细胞的炎症 具有突变的CFTR等位基因的细胞将被描绘出来。粘附性铜绿假单胞菌 刺激上皮细胞表达大量中性粒细胞 趋化因子IL-8。这是通过一个复杂的信令级联来实现的。 上皮细胞对铜绿假单胞菌最早的反应之一是 细胞内钙的快速升高，紧随其后的是 IL-9和其他促炎细胞因子的转录因子。我们 将确定哪些细菌基因产物触发上皮流 钙是引起这种炎症反应所必需的。这个 上皮性受体的性质、激活的激酶和磷酸酶 通过细菌，这一信号通路的调节 已经成立了。内源性刺激，如突变CFTR的积累 在内质网内(内质网超载)也可能触发 细胞内钙类似地通过以下途径启动核因子-kappaB的激活 通过比较具有特定类型CFTR突变的细胞的反应，它 应该可以确定CFTR功能障碍是如何影响免疫的 上皮细胞的功能以及电生理 属性。核因子-kapapB在慢性阻塞性肺疾病细胞中的活化分析 来自转基因小鼠和CF患者的CFTR突变 可能建议采取治疗策略来调节炎症反应。 免疫抑制药如FK506和环孢素A可阻断钙离子 依赖的磷酸酶钙调神经磷酸酶。或许可以使用这种药物来 减轻铜绿假单胞菌引起的炎症，以及 核因子-kappaB对细胞的内源性刺激作用 由突变的、错误的CFTR在小麦中的积累引发的胁迫 内质网。

Cystic fibrosis pulmonary disease is characterized by neutrophil-dominated airway inflammation, which eventually leads to fibrosis, bronchiectasis, and pulmonary failure. Pseudomonas aeruginosa infection is an important cause of this inflammatory process. In the experiments proposed, exactly how this organism stimulates inflammation in both normal and epithelial cells with mutant CFTR alleles will be delineated. Adherent P. aeruginosa stimulate epithelial cells to express large amounts of the neutrophil chemokine IL-8. This is accomplished through a complex signaling cascade. One of the earliest responses of the epithelial cell to P. aeruginosa is a brisk rise in intracellular calcium, which is followed by activation of the transcription factor of IL-9 and other pro-inflammatory cytokines. We will establish which bacterial gene products trigger the epithelial flux in calcium which is required to evoke this inflammatory response. The nature of the epithelial receptor, the kinases and phosphatases activated by the bacteria, and the regulation of this signaling pathway will be established. Endogenous stimuli such as the accumulation of mutant CFTR within the endoplasmic reticulum (ER overload) may also trigger a rise in intracellular calcium which similarly initiates activation of NF-kappaB By comparing the response of cells with specific types of CFTR mutations, it should be possible to determine how CFTR dysfunction affects the immune function of epithelial cells, as well as the electrophysiological properties. Analysis of the activation of NF-kapapB in cells with defined CFTR mutations from both transgenic mice, as well as from patients with CF may suggest therapeutic strategies to modulate the inflammatory response. Immunosuppressant drugs such as FK506 and cyclosporine A block the calcium dependent phosphatase calcineurin. It may be possible to use such drugs to diminish the inflammation provoke by P. aeruginosa as well as the endogenous stimulation of NF-kappaB produced in response to the cell stress initiated by the accumulation of mutant, mistrafficked CFTR in the endoplasmic reticulum.