Human Airway Colonization by Mycoplasma pneumoniae

肺炎支原体在人呼吸道定植

• 批准号: 8786665
• 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/8786665
• 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; 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; public health relevance; 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）：支原体肺炎是社区获得性呼吸道疾病的主要原因，占所有肺炎的20％，占美国每年> 100,000次住院，并且是大儿童和年轻人肺炎的主要原因。感染通常是慢性的，会导致或加剧包括哮喘和COPD在内的改变生命的条件，但是尚不清楚呼吸道的组织逆向和持久性的分子基础。支原体粘附蛋白P1直接在受体结合和滑行运动中起作用。唾液酸糖蛋白和硫化糖脂（如亚磺胺）已被鉴定为受体，但是当与唾液酸糖蛋白结合时，肺炎支原体会滑动，并在与硫酸硫化亚硫酸糖结合时是静态的。我们的长期目标是了解肺炎支原体和气道上皮之间的动态相互作用如何影响传播，组织的向量，主机内传播，免疫逃避和持久性。对潜在的分子识别事件及其如何影响感染结果有更深入的了解将增强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，肺炎支原体粘附和滑行运动受到气道聚糖的密度和结构的影响，这又影响了定殖模式和宿主内传播。令人惊讶的是，尽管气道聚糖的结构和分布对多种呼吸道病原体的根本重要性，但知之甚少。 AIM 1将在潜在的肺炎支原体受体的背景下表征人类气道糖，并在人类气道模型和人类气道组织中发现哪些聚糖的位置和哪些水平。我们将利用最先进的碳水化合物结构分析来定义在空气界面培养物中生长的正常人支气管上皮（NHBE）细胞的糖，我们与之建模了分生孢子定植。我们推测，聚糖环境有助于该模型中观察到的独特的空间和时间定植模式。 NHBE细胞甘油酸分析的结果将与上下气道的人体组织样品的全面硫酸化糖脂分析进行比较。 AIM 2将定义肺炎支原体的结合特异性和力以及对受体种群的滑翔表型，从而解决了潜在受体影响结合和滑翔活性。我们将筛选一个综合的聚糖阵列，以确定结合特异性，通过原子力显微镜确定结合强度，并评估单独和以不同相对浓度组合的单独和组合的唾液糖蛋白和硫化糖脂受体部分的粘附和滑行。 AIM 3将表征粘附素P1蛋白的亚群，该蛋白的特征性单克隆抗体标记模式定义，评估该P1亚群的丰富度的变化与粘附功能和粘附功能相关，并将P1与末端细胞器表面上的结构变化相关联，以及通过电子冰冻层的结构变化。