Biofilm Formation by Pneumocystis

肺孢子菌形成生物膜

• 批准号: 7495414
• 负责人: Melanie T Cushion
• 金额: $ 36.58万
• 依托单位: UNIVERSITY OF CINCINNATI
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-02-01 至 2012-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7495414
• 关键词: Acquired Immunodeficiency Syndrome; Address; Animals; Antimicrobial Resistance; Back; Biochemical; Biological Process; Biology; Cells; Chronic Disease; Chronic Obstructive Airway Disease; Classification; Communicable Diseases; Communication; Compatible; Complex; Condition; Data; Environment; Evaluation; Extracellular Matrix; Gene Expression; Genes; Goals; Growth; Hand; Human; Immune; Immune response; Immune system; Immunocompromised Host; Immunotherapy; In Vitro; Infection; Investigation; Kinetics; Knowledge; Laboratories; Lead; Life Cycle Stages; Lung; Mammals; Metabolic; Methods; Microarray Analysis; Microbe; Microbial Biofilms; Microscopic; Modeling; Molecular; Molecular Profiling; Morbidity - disease rate; Morphology; Mus; Mutation; Natural History; Nature; Numbers; Organism; Outcome; Pathogenicity; Patients; Persons; Plastics; Platelet Factor 4; Pneumocystis; Pneumocystis carinii; Pneumocystis carinii Pneumonia; Pneumonia; Process; Production; Proteins; Public Health; Pulmonary alveolar structure; Rattus; Reporting; Resistance; Resistance to infection; Rodent; Standards of Weights and Measures; Structure; Sulfamethoxazole; Surface; System; Techniques; Testing; Therapeutic Agents; Therapeutic immunosuppression; Trimethoprim-Sulfamethoxazole; Type I Epithelial Receptor Cell; Validation; Work; antimicrobial; assault; concept; experience; fungus; immunosuppressed; in vivo; innovation; knowledge base; member; microbial; microbial community; novel; novel strategies; pathogen; tool; transmission process

DESCRIPTION (provided by applicant): Members of the fungal genus Pneumocystis can cause a lethal pneumonia (PCP) in hosts with debilitated immune systems, such as HIV-infected persons with AIDS; patients undergoing immunosuppressive therapy; and more recently in those patients received targeted immunotherapy. The manner in which the infection is disseminated; the life cycle within the mammalian lung; and the strategies used for survival within this inhospitable environment are largely unknown due in large part to the lack of a continuous cultivation method. It is our long term goal to understand the infection process of Pneumocystis as a means to identify its survival strategies which could then be exploited for interdiction of infection. Production of biofilms is a strategy used by many microbes for protection against environmental assaults; for communication and differentiation among members; and as foci for dissemination. We posit that the attachment and growth of Pneumocystis within the lung alveoli is akin to biofilm formation. An in vitro system was identified that supports apparent biofilm formation by P. carinii (from rat) and P. murina (from mouse). The biofilms showed similar growth kinetics, confocal attributes, and morphological changes compatible with biofilm formation by other fungi. In the present proposal, this system will be optimized to provide a novel tool for understanding the survival strategies of Pneumocystis and then will be used to address questions about the biology of the process. The following specific aims are proposed: (1) Define and characterize the optimal conditions for formation of Pneumocystis biofilms. A systematic approach will be used to evaluate matrices, conditions, and additives leading to robust biofilm formation. Microscopic and quantitative methods will be used to chart the progression of biofilm formation. (2) Identify the biological process associated with biofilm formation. The morphologic changes Pneumocystis undergoes as it forms a biofilm are dramatic. Identification and verification of the genes and gene products involved in the progression using molecular and biochemical methods will provide basic information on the biology of biofilm production and lead to a new appreciation and understanding of this process. (3) Assess the pathogenicity of in vitro biofilms in vivo. The progression of infection initiated by biofilm-derived organisms vs. the standard model will be compared by microscopic and quantitative methods. The in vivo morphology and gene expression will compared to in vitro biofilms for validation of the in vitro system. Optimization and evaluation of the biofilm process will fundamentally advance the study of Pneumocystis. PUBLIC HEALTH RELEVANCE: Microbial pathogens use biofilms to escape the infected host's immune defenses and antimicrobial therapy. We identified a biofilm system for the fungal pathogen, Pneumocystis, that will be used to understand the strategies it employs to survive in the lung and cause pneumonia.

描述（由申请人提供）：肺孢子菌属真菌的成员可在免疫系统衰弱的宿主中引起致死性肺炎（PCP），例如HIV感染的AIDS患者;接受免疫抑制治疗的患者;以及最近接受靶向免疫治疗的患者。感染传播的方式;哺乳动物肺内的生命周期;以及在这种不适宜居住的环境中用于生存的策略在很大程度上是未知的，这在很大程度上是由于缺乏连续的培养方法。我们的长远目标是了解肺孢子虫的感染过程，以找出其生存策略，从而阻断感染。生物膜的产生是许多微生物用于保护免受环境攻击的策略;用于成员之间的交流和分化;以及作为传播的焦点。我们认为肺孢子虫在肺泡内的附着和生长类似于生物膜的形成。鉴定了支持卡氏肺孢子虫（来自大鼠）和鼠肺孢子虫（来自小鼠）的明显生物膜形成的体外系统。的生物膜表现出类似的生长动力学，共聚焦属性，和形态学变化与其他真菌的生物膜形成兼容。在本提案中，该系统将被优化，以提供一个新的工具，了解肺孢子虫的生存策略，然后将被用来解决有关的生物学过程的问题。具体目标如下：（1）确定并表征肺孢子虫生物膜形成的最佳条件。将使用系统方法来评价导致稳健生物膜形成的基质、条件和添加剂。将使用显微镜和定量方法来绘制生物膜形成的进展。(2)识别与生物膜形成相关的生物过程。肺孢子虫在形成生物膜时所经历的形态变化是戏剧性的。使用分子和生物化学方法鉴定和验证参与进展的基因和基因产物将提供生物膜生产生物学的基本信息，并导致对该过程的新的认识和理解。(3)评估体外生物膜在体内的致病性。将通过显微镜和定量方法比较生物膜衍生微生物与标准模型引发的感染进展。将体内形态和基因表达与体外生物膜进行比较，以验证体外系统。生物膜法的优化和评价将从根本上推进肺孢子虫的研究。公共卫生相关性：微生物病原体利用生物膜逃避感染宿主的免疫防御和抗菌治疗。我们确定了一个生物膜系统的真菌病原体，肺孢子虫，这将被用来了解它采用的策略，在肺部生存，并导致肺炎。