Rhinoviruses, Epithelial Cells, and Airway Function

本应用程序的目的是检查鼻病毒（RV）感染如何诱导哮喘恶化。这些研究解决了流行病学研究之间的不一致，流行病学研究表明RV感染是哮喘加重的常见诱因，而临床研究表明接种哮喘受试者总是引起“感冒”症状，有时会引起肺功能和下气道炎症的细微变化，但很少引起实际的哮喘加重。我们的中心假设是，RV感染反应的性质和强度是由感染菌株和/或被感染上皮细胞的固有特性决定的。项目1 （J Widdicombe，项目负责人）将研究不同RV菌株感染对在气液界面培养的具有良好特征的气道上皮细胞的结构、功能和介质产生的影响。本项目还将研究从患有和不患有过敏性气道疾病的人身上获得的鼻腔和支气管上皮细胞培养物，从患有和不患有过敏性气道疾病的人身上获得的鼻腔和支气管上皮细胞培养物，以及从对RV感染有不同临床反应的人身上获得的培养物对RV感染的反应。项目2 （H. Boushey）将通过培养急性哮喘发作患者和前瞻性哮喘受试者的鼻分泌物来获得RV。我们还将描述上呼吸道和下呼吸道对感染的临床和炎症反应。本项目将进一步比较接种两株RV16的临床反应，这两株RV16对上皮细胞的体外作用不同。它还将提供（项目1）来自过敏性气道疾病和RV感染反应性受试者的鼻和支气管上皮细胞。项目（R. WU）将对RV16感染支气管上皮细胞引起的基因表达变化进行阵列研究，并比较相近的RV16菌株在体外和临床效果不同时引起的基因表达变化模式。病毒学核心（D. Schnurr）将培养、分离、鉴定和滴定从受感染对象或上皮细胞中获得的RV，将连续传代RV16以研究菌株特异性感染效应，并将对这些菌株进行测序，以确定菌株特异性活性差异的遗传位点。定量遗传学核心（C. Dolganolv）将在三个项目中对体外和体内感染的上皮细胞中大量选定基因的变化进行量化。通过这种综合方法，我们希望确定RV感染对哮喘患者影响的关键决定因素，从而提出潜在的预防和治疗新方法。

The goal of this application is to examine how rhinovirus (RV) infection induces worsening of asthma. The studies address the discordance between epidemiologic studies showing that RV infection is a common precipitant of asthma exacerbations and clinical studies showing that RV inoculation of asthmatic subjects consistently causes symptoms of a "cold." sometimes cause subtle changes in pulmonary function and in lower airway inflammation, but rarely causes actual asthma exacerbations. Our central hypothesis is that the nature and intensity of the response to RV infection is determined by properties inherent to the infecting strain and/or to the epithelial cells infected. Project 1 (J Widdicombe, Project Leader) will examine the effects of infection with different strains of RV on the structure, function, and mediator production of well-characterized airway epithelial cells cultured at an air-liquid interface. This project will also examine the response to RV infection of cultures of nasal and bronchial epithelial cells obtained from people with and without allergic airway disease, and from nasal and bronchial epithelial cells obtained from people with and without allergic airway disease, and from people with different clinical responses to RV infection. Project 2 (H. Boushey) will obtain RV's by culturing nasal secretions from patients with acute asthma exacerbations and from asthmatic subjects followed prospectively. We will also characterize the clinical and inflammatory responses of the upper and lower airways to infection. This project will further compare the clinical responses to inoculation with two strains of RV16 that differ in their in vitro effects on epithelial cells. It will also provide (to Project 1) nasal and bronchial epithelial cells from subjects characterized as to allergic airway disease and responsiveness to RV infection. Project (R. WU) will conduct array studies of changes in gene expression induced by RV16 infection of bronchial epithelial cells and will compare the patterns of change induced by closely related strains of RV that differ in their in vitro and clinical effects. The Virology Core (D. Schnurr) will culture, isolate, identify, and titrate RV's obtained from infected subjects or epithelial cells, will serially passage RV16 for studies of strain-specific effects of infection, and will sequence these strains to identify genetic sites accounting for strain-specific differences in activity. The Quantitative Genetics Core (C. Dolganolv) will quantify changes in large numbers of selected genes in epithelial cells infected in vitro and in vivo in the three projects proposed. Through this integrated approach, we hope to identify critical determinants of the effects of RV infection in people with asthma, and thus to suggest potential new approaches to prevention and treatment.