MAP KINASE ACTIVATION IN RSV-INDUCED CHEMOKINE SECRETION

描述（改编自申请人的摘要）：Casola博士感兴趣的是 在病毒感染期间诱导促炎基因的机制， 可以有针对性地开发新的方法来调节两者 体内炎症和免疫反应。RSV是最常见的原因， 儿童流行性呼吸道疾病。引发的炎症反应 由呼吸道上皮细胞感染，是一种重要的致病性 RSV疾病的组成部分。白细胞介素8（IL-8）是一种CXC趋化因子， RSV感染者鼻和支气管肺泡灌洗液中的高浓度 儿童可能在炎症细胞的招募中发挥作用， RSV感染后的肺部。呼吸道上皮细胞中的RSV复制 在体外导致IL-8基因表达和核因子κ B增加 （NF-kB）活化。本项目将继续研究RSV诱导 IL-8基因表达部分通过丝裂原活化的 蛋白激酶（MAPK）级联反应。MAPK细胞外调节的刺激 激酶（ERK 1/2）和p38导致转录因子的激活， 调节IL-8基因转录。三个目标旨在测试 假说. 首先，她将研究Raf-mitogen的激活 呼吸道合胞病毒感染中活化的细胞外调节激酶（MEK）-ERK通路 及其在RSV诱导的IL-8分泌中的作用。ERK 1/2的作用机制 活化将通过评估所述细胞的激酶活性来确定。 IL-8基因上游分子Raf及各激酶的作用 诱导，在转录因子活化和IL-8蛋白合成中的作用 显性失活突变体的过表达或通过下调 反义寡核苷酸其次，她将调查 MAP激酶p38和c-JUN-N-末端激酶（JNK）在RSV感染中作用， 它们在RSV诱导的IL-8分泌中的作用。与目标1类似的方法 将用于确定MAPK级联反应的这两个分支的作用 RSV诱导的IL-8产生。 最后，她将集中精力建立 MAPK级联在刺激特异性NF-κ B激活机制中的作用 通过RSV感染和肿瘤坏死因子（TNF）刺激，诱导 不同的翻译后修饰的NF-κ B和是否 IL-8启动子共激活因子的募集和MAPK的作用 将研究这些过程中的级联。这些研究可以识别 参与病毒诱导的趋化因子IL-8的细胞内信号传导途径 生产，可用于开发新的治疗策略， 急性肺部炎症和感染后哮喘的治疗。

DESCRIPTION (adapted from the applicant's abstract): Dr. Casola is interested in mechanisms of induction of pro-inflammatory genes during viral infections that could be targeted to develop novel approaches for modulation of both inflammatory and immune responses in vivo. RSV is the most common cause of epidemic respiratory disease in children. The inflammatory response, triggered by the infection of respiratory epithelial cells, is an essential pathogenic component of RSV disease. Interleukin 8 (IL-8) is a CXC chemokine detected in high concentrations in the nasal and bronchoalveolar lavages of RSV-infected children that could play a role in the recruitment of inflammatory cells to the lung following RSV infection. RSV replication in airway epithelial cells in vitro results in increased IL-8 gene expression and nuclear factor kappa B (NF-kB) activation. This project will pursue the hypothesis that RSV induces IL-8 gene expression partially through the activation of the mitogen-activated protein kinase (MAPK) cascade. Stimulation of MAPK extracellular regulated kinase (ERK1/2) and p38 results in activation of transcription factors that regulate IL-8 gene transcription. Three aims are designed to test the hypothesis. First, she will investigate the activation of Raf-mitogen activated extracellular regulated kinase (MEK)-ERK pathway in RSV infection and its role in RSV-induced secretion of IL-8. The mechanism(s) of ERK1/2 activation will be determined by evaluating the kinase activity of the upstream molecule Raf and the role of each individual kinase in IL-8 gene induction, in transcription factor activation and in IL-8 protein synthesis by overexpression of dominant negative mutants or by down regulation with antisense oligonucleotides. Secondly, she will investigate the activation of the MAP kinase p38 and c-JUN-N-terminal kinase (JNK) in RSV infection and their role in RSV-induced secretion of IL-8. A similar approach as in Aim 1 will be used to establish the role of these two branches of the MAPK cascade in RSV-induced IL-8 production. Finally, she will focus on establishing the role of the MAPK cascade in a stimulus-specific mechanism of NF-kB activation by RSV infection and tumor necrosis factor (TNF) stimulation, the induction of different post-translational modifications of NF-kB and whether the recruitment of co-activators to the IL-8 promoter, and the role of MAPK cascade in these processes will be investigated. These studies may identify intracellular signaling pathways involved in the viral-induced chemokine IL-8 production that could be used to develop new therapeutic strategies useful in the treatment of acute lung inflammation and post-infectious asthma.