Chemokine and Protein Patterns in RSV Infection

RSV 感染中的趋化因子和蛋白质模式

• 批准号: 6878399
• 负责人: Roberto P Garofalo
• 金额: $ 16.94万
• 依托单位: UNIVERSITY OF TEXAS MED BR GALVESTON
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-01 至 2010-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6878399
• 关键词: SDS polyacrylamide gel electrophoresis; asthma; cell migration; cellular immunity; chemokine; cytotoxic T lymphocyte; genetically modified animals; high throughput technology; host organism interaction; immunopathology; inflammation; laboratory mouse; leukocyte activation /transformation; lung disorder; macrophage inflammatory proteins; mass spectrometry; matrix assisted laser desorption ionization; molecular biology information system; nasopharynx; natural killer cells; protein structure function; respiratory syncytial virus; virus infection mechanism; virus protein; virus replication

Respiratory Syncytial Virus (RSV) infections have been uniquely linked to the development and the severity of asthma. Virus-specific cellular immunity, particularly the CTL response, is implicated in the immunopathogenesis of RSV infection. We have shown that nasopharyngeal concentrations of MIP-1alpha, a prototype of viral-inducible airway epithelial chemokines, correlate with the degree of illness severity in RSV-infected children and that mice genetically deficient in MIP-1 alpha (-/-) have a striking reduction in lung inflammation. We hypothesize that MIP-alpha, by virtue of its activity on both NK cells and CTL, functions as a bridge between innate and adaptive immunity to RSV, thus playing a crucial role in restricting viral replication, yet inducing mucosal inflammation, wheezing and airway hyperresponsiveness (AHR). In this project we propose the following aims: 1. Identify the contribution of MIP-1 alpha in the control of viral replication and development of RSV-induced lung inflammation, AHR and illness. Using MIP-1alpha -/- mice, we will test the hypothesis that MIP-let expression is necessary for viral clearance, but also linked to the pathogenesis of AHR and clinical illness in RSV infections. 2. Investigate the requirement of MIP-1 et for the migration and activation of NK cells and NK-cell driven CTL responses in RSV infection. We will test whether MIP-1alpha promotes NK cell migration to the lung, activation, and antiviral function and whether it regulates NK-dependent RSV-specific CTL responses. 3. Analyze the spectrum of RSV-inducible proteins in the lung of mice, either control or MIP-1 alpha deficient, using a high-throughput proteomics approach with 2D SDS-PAGE and MALDI-TOF mass spectroscopy. We will generate databases of lung proteins from RSV-infected mice to identify downstream proteins/mediators affected by MIP-let-dependent pathways. 4. Analyze whether distinct protein patterns at the airway mucosal site can discriminate between infants with different severity of illness or degree of chemokine response following naturally-acquired RSV infection. By the high-throughput proteomics approach we will identify specific proteins or protein patterns in nasopharyngeal secretions that may contribute to the pathogenesis or severity of RSV-induced disease, and are associated with greater production of MIP-1alpha or other epithelial-derived chemokines. These studies will contribute to the identification of new strategies to promptly recognize, prevent, or early treat the most severe clinical forms of RSV infection in infancy, thus reducing the long-term burden of recurrent wheezing and asthma.

呼吸道合胞病毒（RSV）感染与哮喘的发展和严重程度有着独特的联系。病毒特异性细胞免疫，特别是CTL应答，与RSV感染的免疫发病机制有关。我们已经证明，鼻咽部MIP-1 α（病毒诱导的气道上皮趋化因子的原型）的浓度与RSV感染儿童的疾病严重程度相关，并且MIP-1 α（-/-）基因缺陷的小鼠肺部炎症显著减少。我们假设MIP-α，凭借其对NK细胞和CTL的活性，作为对RSV的先天性和适应性免疫之间的桥梁，从而在限制病毒复制，但诱导粘膜炎症，喘息和气道高反应性（AHR）中发挥关键作用。在这个项目中，我们提出了以下目标：1。确定MIP-1 α在控制病毒复制和RSV诱导的肺部炎症、AHR和疾病发展中的作用。使用MIP-1 α-/-小鼠，我们将检验MIP-let表达对于病毒清除是必要的，但也与AHR的发病机制和RSV感染的临床疾病有关的假设。2.研究MIP-1 et对RSV感染中NK细胞迁移和活化以及NK细胞驱动的CTL应答的需求。我们将测试MIP-1 α是否促进NK细胞迁移到肺部，激活和抗病毒功能，以及它是否调节NK依赖的RSV特异性CTL应答。3.分析RSV诱导蛋白质的谱， 使用具有2D SDS-PAGE和MALDI-TOF质谱的高通量蛋白质组学方法，对对照或MIP-1 α缺陷的小鼠的肺进行分析。我们将从RSV感染的小鼠中生成肺蛋白的数据库，以识别受MIP-let依赖性途径影响的下游蛋白/介质。4. 分析气道粘膜部位的不同蛋白质模式是否可以区分自然获得性RSV感染后疾病严重程度或趋化因子反应程度不同的婴儿。通过高通量 通过蛋白质组学方法，我们将鉴定鼻咽分泌物中可能与RSV诱导疾病的发病机制或严重程度有关的特定蛋白质或蛋白质模式，并与MIP-1 α或其他上皮衍生趋化因子的大量产生有关。这些研究将有助于确定新的策略，以及时识别，预防或早期治疗婴儿期最严重的RSV感染临床形式，从而减少复发性喘息和哮喘的长期负担。