MAP KINASE ACTIVATION IN RSV-INDUCED CHEMOKINE SECRETION

描述(改编自申请者的摘要)：卡索拉博士对 在病毒感染过程中诱导促炎基因的机制 可以针对这两种情况开发新的调制方法 体内的炎症和免疫反应。呼吸道合胞病毒是引起 儿童中流行的呼吸道疾病。引发的炎症反应 由呼吸道上皮细胞感染，是一种必不可少的致病因素 呼吸道合胞病毒病的组成部分。白介素8是一种CXC趋化因子，在 呼吸道合胞病毒感染患者鼻腔和支气管肺泡灌洗液中的高浓度 儿童可能在招募炎性细胞方面发挥作用 呼吸道合胞病毒感染后的肺。呼吸道合胞病毒在呼吸道上皮细胞中的复制 体外结果增加IL-8基因表达和核因子kappaB (核因子-kB)激活。这个项目将追寻RSV诱导的假说 IL-8基因表达部分通过丝裂原激活 蛋白激酶(MAPK)级联反应。细胞外调节MAPK的刺激作用 激酶(ERK1/2)和p38导致转录因子激活，从而 调节IL-8基因转录。设计了三个目标来测试 假设。首先，她将研究Raf-有丝分裂原的激活 呼吸道合胞病毒感染中激活的细胞外调节激酶-ERK信号转导途径 以及它在RSV诱导的IL-8分泌中的作用。ERK1/2的作用机制(S) 激活将通过评估蛋白的激酶活性来确定。 上游分子Raf及其在IL-8基因中的作用 诱导转录因子激活和IL-8蛋白合成 显性负性突变体的过度表达或通过下调 反义寡核苷酸。其次，她将调查激活的 呼吸道合胞病毒感染与MAP激酶p38和c-jun-N末端激酶(JNK)的关系 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级联的这两个分支的作用 在RSV诱导的IL-8产生中。最后，她将专注于建立 MAPK级联信号通路在核因子-kB激活的刺激特异性机制中的作用 通过RSV感染和肿瘤坏死因子(TNF)刺激，诱导 核因子-kB的不同翻译后修饰以及是否 IL-8启动子共激活因子的募集及MAPK的作用 将对这些过程中的级联进行调查。这些研究可能会确定 病毒诱导趋化因子IL-8的细胞内信号转导途径 可用于开发新的治疗策略的产品 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.