Human Airway Colonization by Mycoplasma pneumoniae

肺炎支原体在人呼吸道定植

• 批准号: 8849366
• 负责人: DUNCAN C KRAUSE
• 金额: $ 38.91万
• 依托单位: UNIVERSITY OF GEORGIA
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-05-15 至 2019-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8849366
• 关键词: Accounting; Address; Adherence; Air; Asthma; Atomic Force Microscopy; Bacterial Adhesins; Binding; Carbohydrates; Cells; Characteristics; Chemotaxis; Child; Chronic; Chronic Obstructive Airway Disease; Cilia; Communities; Complement Receptor; Data; Electrons; Environment; Epithelial Cells; Event; Gel; Glycoproteins; Glycosphingolipids; Goals; Health; Histocytochemistry; Hospitalization; Human; Immune; In Vitro; Incidence; Infection; Inorganic Sulfates; Knowledge; Label; Lectin; Life; Link; Liquid substance; Lung diseases; Mediating; Modeling; Molecular; Monoclonal Antibodies; Mucins; Mycoplasma; Mycoplasma pneumoniae; Nature; Organelles; Outcome; Pathogenesis; Pattern; Phenotype; Pneumonia; Polysaccharides; Population; Preparation; Process; Proteins; Publishing; Relative (related person); Sialic Acids; Sialoglycoproteins; Source; Specificity; Structure; Sulfoglycosphingolipids; Surface; Testing; Tissue Sample; Tissues; Unspecified or Sulfate Ion Sulfates; Work; airway epithelium; base; cell motility; density; glycolipid receptor; human tissue; interest; molecular recognition; pathogen; receptor; receptor binding; respiratory; screening; sulfoglycolipids; tissue tropism; transmission process; young adult

DESCRIPTION (provided by applicant): Mycoplasma pneumoniae is a major cause of community-acquired respiratory disease, accounting for >20% of all pneumonia and >100,000 hospitalizations annually in the U.S., and is the leading cause of pneumonia in older children and young adults. Infections are often chronic and can precipitate or exacerbate life-altering conditions including asthma and COPD, but the molecular basis for tissue tropism and persistence in the airways is not known. Mycoplasma adhesin protein P1 functions directly in receptor binding and gliding motility. Sialoglycoproteins and sulfated glycolipids such as sulfatide have been identified as receptors, but M. pneumoniae cells glide when bound to sialoglycoproteins and are static when bound to sulfatide. Our long- term goal is to understand how the dynamic interplay between M. pneumoniae and airway epithelium impacts transmission, tissue tropism, within-host spread, immune evasion, and persistence. A greater understanding of the underlying molecular recognition events and how they impact infection outcome will enhance our ability to develop new strategies to limit incidence and impact. Our central hypothesis is that M. pneumoniae adherence and gliding motility are influenced by the density and structure of airway glycans, which in turn impacts colonization pattern and within-host spread. Surprisingly little is known about the structure and distribution of airway glycans, despite their fundamental importance for diverse respiratory pathogens. Aim 1 will characterize the human airway glycome in the context of potential M. pneumoniae receptors, addressing which glycans are found where and at what levels in a human airway model and in human airway tissues. We will utilize state-of- the-art carbohydrate structural analysis to define the glycome of normal human bronchial epithelial (NHBE) cells grown in air-liquid interface culture, with which we model mycoplasma colonization. We speculate that glycan environment contributes to the distinct spatial and temporal colonization patterns observed in this model. Results from NHBE cell glycome analysis will be compared with comprehensive sulfated glycolipid analysis of human tissue samples from upper and lower airways. Aim 2 will define M. pneumoniae binding specificity and force and gliding phenotype on receptor populations, addressing which potential receptors impact binding and gliding activity. We will screen a comprehensive glycan array to determine binding specificity, determine binding strength by atomic force microscopy, and evaluate adherence and gliding on sialoglycoprotein and sulfated glycolipid receptor moieties individually and in combination at different relative concentrations. Aim 3 will characterize a subpopulation of the adhesin P1 protein that is defined by its characteristic monoclonal antibody labeling pattern, assess how changes in the abundance of this P1 subpopulation correlate with adherence and gliding function, and correlate P1 dynamics with structural changes on the terminal organelle surface, as observed by electron cryotomography.

描述（由申请人提供）：肺炎支原体是社区获得性呼吸道疾病的主要原因，在美国每年占所有肺炎的>20%和> 100，000例住院治疗，并且是年龄较大的儿童和年轻人肺炎的主要原因。感染通常是慢性的，可以加速或加剧改变生活的疾病，包括哮喘和COPD，但组织嗜性和气道持久性的分子基础尚不清楚。支原体粘附素蛋白P1直接在受体结合和滑动运动中起作用。唾液酸糖蛋白和硫酸化糖脂如硫苷脂已被确定为受体，但M。当与唾液酸糖蛋白结合时，肺炎杆菌细胞滑动，当与硫苷脂结合时，肺炎杆菌细胞静止。我们的长期目标是了解M之间的动态相互作用。肺炎和呼吸道上皮细胞影响传播、组织嗜性、宿主内传播、免疫逃避和持久性。更好地了解潜在的分子识别事件及其如何影响感染结果将提高我们制定新策略以限制发病率和影响的能力。我们的中心假设是M.肺炎链球菌的粘附和滑行运动受到气道聚糖的密度和结构的影响，这反过来又影响定植模式和宿主内传播。令人惊讶的是，对气道聚糖的结构和分布知之甚少，尽管它们对不同的呼吸道病原体至关重要。目的1将在潜在M的背景下表征人气道糖组。肺炎链球菌受体，解决了在人气道模型和人气道组织中在何处和以何种水平发现哪些聚糖。我们将利用最先进的碳水化合物结构分析来确定在气-液界面培养中生长的正常人支气管上皮（NHBE）细胞的糖组，我们用它来模拟支原体定植。我们推测，聚糖环境有助于在这个模型中观察到的不同的空间和时间的殖民模式。将NHBE细胞糖组分析的结果与来自上呼吸道和下呼吸道的人体组织样本的综合硫酸化糖脂分析进行比较。目标2将定义M。本发明的目的在于提供一种用于测定肺炎链球菌结合特异性和力以及受体群体上的滑动表型的方法，解决了哪些潜在受体影响结合和滑动活性。我们将筛选一个全面的聚糖阵列，以确定结合特异性，通过原子力显微镜确定结合强度，并评估唾液酸糖蛋白和硫酸化糖脂受体部分的粘附和滑动，单独和组合在不同的相对浓度。目的3将表征粘附素P1蛋白的亚群，该亚群由其特征性单克隆抗体标记模式定义，评估该P1亚群的丰度变化如何与粘附和滑动功能相关，并将P1动态与末端细胞器表面的结构变化相关，如通过电子冷冻断层扫描观察到的。