Mucosal Immune Barrier in Infection and Immunity

感染和免疫中的粘膜免疫屏障

• 批准号: 7690382
• 负责人: Keith E Mostov
• 金额: $ 130万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-30 至 2013-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7690382
• 关键词: Mucosal; Immune; Barrier; Infection; Immunity

DESCRIPTION (provided by applicant): Overview of Program Project: This is a resubmission of a renewal of a Program Project Grant (PPG) application to study the mucosal barrier in infection and inflammation. Ninety-five percent of infectious agents enter through exposed mucosal surfaces, such as the respiratory, gastrointestinal, mammary and urogenital tracts. Mucosal infections include bacterial and viral pneumonia, SARS, TB, AIDS and other sexually transmitted diseases, numerous opportunistic, emerging and re-emerging infections, and biological warfare/terrorist agents. Mucosal surfaces are lined by epithelial cells, usually in a monolayer. The epithelial cell layer is the principal barrier to entry of infectious agents, allergens and other noxious antigens. Fundamentally, the epithelial layer is the most basic component of the innate mucosal immune system. In this Program Project we focus on two broad and inter-related areas of how the epithelial layer performs its functions in mucosal immune protection, with a particular emphasis on pulmonary epithelium. Projects 1 (Mostov), 2 (Engel) and 3 (Rosen) study mechanisms of epithelial wound healing, the roles of sulfs (sulfatases which modify heparan sulfate proteoglycans) in epithelial response to injury and the interaction of polarized cells with a bacterial pathogen. Project 4 (Werb) is focused on how inflammatory cells cross the epithelial barrier to enter the lumen of the organ. Project 1 uses two novel three dimensional cell culture models of epithelial wound healing. Project 2 focuses on the opportunistic pathogen, Pseudomonas aeruginosa, as a model for epithelial pathogen interaction. Project 3 examines how sulfs participate in epithelial wound healing. Project 4 focuses on the role of proteases and sulfs in regulating inflammatory cell recruitment and migration in mucosal tissues. Core A supports the Program Project administratively. Core B (Matthay) provides primary human and rodent lung epithelial cells in monolayer cultures, three-dimensional models and lung slices, as well as functional studies in mice. Core C has an array of four microscopes suitable for three- and four-dimensional analysis of cell dynamics for in vitro studies as well as experiments in living, anesthetized mice. Most infectious agents enter the body through exposed mucosal surfaces. These surfaces are lined by epithelial cells, most commonly in a single layer. We are studying how this epithelial barrier protects us against infection. PROJECT 1: Epithelial wound healing in 3 dimensions (Mostov, K) Project 1 Description (provided by applicant): Greater than 95% of infectious agents enter through exposed mucosal surfaces, such as the respiratory, gastrointestinal and genitourinary tracts. These include HIV, sexually transmitted diseases, numerous opportunistic infections, TB, many emerging and re-emerging infections, and biological warfare/terrorist agents, such as anthrax, Yersinia pestis and smallpox. Most mucosal surfaces are lined by a monolayer of polarized epithelial cells, which forms the principal barrier to entry by infectious agents. In essence, the epithelial layer can be considered the most basic component of the innate mucosal immune system. Some pathogens cross the epithelial layer by disrupting it. Other pathogens exploit disruptions in the monolayer, which can be caused by tissue injury secondary to inflammation, trauma, or may result from cell death or division within the monolayer. To maintain their function as a barrier to infection, epithelial tissues have developed efficient wound healing mechanisms. Wound healing is central to mucosal defense against infection. The epithelial barrier must be restored as quickly as possible, to minimize the opportunity for entry of infectious agents. Some infectious agents, such as Pseudomonas aeruginosa, not only exploit pre-existing wounds, but also impede the wound healing process. We are studying epithelial wound healing by growing epithelial cells in three-dimensional cultures of extracellular matrix, which causes the cells to more closely resemble in vivo conditions. In Aim 1 we are using a system of human primary lung alveolar type II cells, which form alveolar-like cysts, as a model to study response to acute lung injury/acute respiratory distress syndrome. In Aim 2, we are using a three-dimensional system of a well-differentiated human airway cell line, which forms cysts and tubules lined by pseudostratified epithelium, as a model to study airway response to injury. In Aim 3 we are studying the roles of matrix metalloproteinases and sulfatases (enzymes that remove 6-O-sulfate groups from heparan sulfate proteoglycans) in our three dimensional culture systems. This work will be in collaboration with Projects 2, 3 and 4, and supported by Cores B and C. Most infectious agents enter through the layer of cells that lines internal organs, such as the lung. Injuries to this cell layer make it much easier for pathogens to enter and we are studying how the cell layer heals itself.

描述（由申请人提供）：项目概述：这是项目项目拨款（PPG）申请的续期申请，用于研究感染和炎症中的粘膜屏障。95%的感染源通过暴露的粘膜表面进入，如呼吸道、胃肠道、乳腺和泌尿生殖道。粘膜感染包括细菌性和病毒性肺炎，SARS，结核病，艾滋病和其他性传播疾病，许多机会性的，新出现和再出现的感染，以及生物战/恐怖主义代理人。粘膜表面由上皮细胞排列，通常为单层。上皮细胞层是感染因子、过敏原和其他有害抗原进入的主要屏障。从根本上说，上皮是先天粘膜免疫系统最基本的组成部分。在本项目中，我们重点关注上皮层如何在粘膜免疫保护中发挥其功能的两个广泛且相互关联的领域，特别强调肺上皮。项目1 （Mostov）、2 （Engel）和3 （Rosen）研究了上皮伤口愈合的机制、硫酸盐（修饰硫酸肝素蛋白聚糖的硫酸酯酶）在上皮损伤反应中的作用以及极化细胞与细菌病原体的相互作用。项目4 （Werb）的重点是炎症细胞如何穿过上皮屏障进入器官的管腔。项目1使用了两种新颖的上皮性伤口愈合的三维细胞培养模型。项目2的重点是机会致病菌铜绿假单胞菌，作为上皮病原体相互作用的模型。项目3研究硫如何参与上皮性伤口愈合。项目4主要研究蛋白酶和硫在调节粘膜组织炎症细胞募集和迁移中的作用。核心A在行政上支持计划项目。Core B （Matthay）提供人类和啮齿动物肺上皮细胞的单层培养、三维模型和肺切片，以及小鼠的功能研究。Core C有四个显微镜阵列，适用于体外研究和活体麻醉小鼠实验的三维和四维细胞动力学分析。大多数传染性病原体通过暴露的粘膜表面进入人体。这些表面由上皮细胞排列，最常见的是单层。我们正在研究这种上皮屏障如何保护我们免受感染。