Phagocyte Contact Response in Aspergillosis Immunity

曲霉病免疫中的吞噬细胞接触反应

• 批准号: 6836541
• 负责人: JAMES B BURRITT
• 金额: $ 7.08万
• 依托单位: MONTANA STATE UNIVERSITY - BOZEMAN
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-01-01 至 2006-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6836541
• 关键词: Aspergillus flavus; NAD(P)H dehydrogenase; alveolar macrophages; aspergillosis; cellular immunity; clinical research; diagnostic respiratory lavage; disease /disorder model; flow cytometry; genetically modified animals; host organism interaction; human subject; immunocytochemistry; laboratory mouse; lung; microorganism immunology; microorganism interaction; molecular dynamics; monoclonal antibody; neutrophil; opportunistic infections; peptide library; phage display; phagocytes; western blottings

DESCRIPTION (provided by applicant): Aspergillus fumigatus is the leading airborne fungal pathogen in immune compromised people. Due to difficulties in managing A. fumigatus infections, most individuals that develop invasive aspergillosis will die. With an increasing immune compromised population, infections by A. fumigatus will also increase. While significant advancements in understanding aspergillosis have been made, the relationship among corresponding immune cells is not fully understood. The long-term goal of this work is to identify the molecular basis of host defense against A. fumigatus. The specific focus is to determine the cellular and molecular contact between phagocytes and A. fumigatus and the role of microbicidal responses in the corresponding immunity. Three aims are proposed to address this goal: 1) To evaluate the cell specific response to A. fumigatus conidia in the lungs of mice by flow cytometry and immunohistochemistry: Lung tissue will be probed using specific monoclonal antibodies to identify cells that arrive following inoculation of A. fumigatus conidia in normal and immune suppressed mice. Both cortisone-induced immune suppression and mice made susceptible to by knockout of the NADPH oxidase will be examined. 2) To investigate the role of the NADPH oxidase in subcellular compartments of immune cells which have ingested conidia: Superoxide generation assays will be used to show assembly of the oxidase associated with phagolysosomes containing ingested conidia. The assembled complex will be confirmed by demonstrating superoxide generation rates in compartments containing conidia, as well as demonstrating the complete set of oxidase components by immunoblot. A panel of monoclonal antibodies is available for oxidase detection. 3) To identify unique peptides that bind specifically to the A. fumigatus conidia and hyphal forms by phage-display peptide library analysis. Affinity selection of unique peptides will be carried out by probing the A. fumigatus resting conidia, swollen conidia, and hyphae with a peptide library. This information will be used to suggest mechanisms of attachment in this host-pathogen relationship

描述（由申请人提供）： 烟曲霉菌是免疫功能低下人群中主要的空气传播真菌病原体。由于管理A.烟曲霉感染，大多数发展为侵袭性曲霉病的个体将死亡。随着免疫力低下人群的增加，A.烟曲霉也会增加。虽然在了解曲霉病方面取得了重大进展，但相应免疫细胞之间的关系尚未完全了解。本工作的长期目标是确定宿主防御A.烟熏。具体的重点是确定吞噬细胞和A之间的细胞和分子接触。以及杀微生物反应在相应免疫中的作用。本研究提出了三个目标：1）评价细胞对A.通过流式细胞术和免疫组织化学检测小鼠肺中的烟曲霉分生孢子：使用特异性单克隆抗体探测肺组织以鉴定接种烟曲霉后到达的细胞。烟曲霉分生孢子在正常和免疫抑制小鼠中的分布。将检查可的松诱导的免疫抑制和通过敲除NADPH氧化酶使小鼠易感的免疫抑制。2)为了研究NADPH氧化酶在已摄入分生孢子的免疫细胞的亚细胞区室中的作用：将使用超氧化物生成测定来显示与含有摄入分生孢子的吞噬溶酶体相关的氧化酶的组装。组装的复合物将通过证明含有分生孢子的隔室中的超氧化物生成速率以及通过免疫印迹证明整套氧化酶组分来确认。一组单克隆抗体可用于氧化酶检测。3)鉴定特异性结合A.通过噬菌体展示肽库分析烟曲霉分生孢子和菌丝形态。独特肽的亲和力选择将通过探测A.烟曲霉休眠分生孢子、肿胀分生孢子和菌丝的肽文库。这一信息将被用来提出在这种宿主-病原体关系中的附着机制