Tools for Analysis of Airway Inflammation

该计划项目的一个统一主题是专注于进化的细胞和分子研究 呼吸道炎症的微环境。这三个项目都利用了遗传老鼠模型。肺支原体感染小鼠作为慢性呼吸道炎症的模型，以及对呼吸道和肺内细胞和组织的尖端双光子和共聚焦成像。为此，由联席主任共同管理的鼠标工具核心将通过提供三个主要功能来服务于项目的共同需求：(I)标准化肺炎支原体的准备、感染程序和感染小鼠的处理；(Ii)开发、优化和实施用于识别突变小鼠的基因分型程序；以及(Iii)开发和提供对参与呼吸道和肺部炎症发展的细胞的实时分析的尖端方法。首先，核心工作人员将生产有毒的肺支原体生物体，这些生物体将在所有实验中进行测试，以确保以最小的变异性提供一致的供应。其次，该中心将使用标准化的流式细胞术、DNA制备和聚合酶链式反应程序，从将用于这三个项目的突变群体中对小鼠进行基因分型。第三, 该核心将通过与加州大学旧金山分校生物成像开发中心(www.ucsf edu/bitc/)的合作，提供对新开发的技术的访问和帮助，以实时查看呼吸道和肺中的活细胞。集中和协调感染、基因分型程序和成像方法将避免资源重复，确保所有群体使用标准化程序，促进信息交换，并促进协作互动。

A unifying theme of the Program Project is the focus on cellular and molecular studies of the evolving microenvironment of airway inflammation. All three projects take advantage of genetic mouse models. Mycoplasma pulmonis infected mice as a model of chronic airway inflammation, and cutting-edge 2-photon and confocal imaging of cells and tissues in the ainways and lung. To this end, the mouse tools core, jointly administered by the co-directors, will serve the common needs of the projects by providing three main functions: (i) to standardize M. pulmonis preparation, infection procedures, and handling of infected mice; (ii) to develop, optimize and implement genotyping procedures for identifying mutant mice; and (iii) to develop and provide access to cutting edge methods for the real-time analysis of cells involved in the development of inflammation in the ainways and lung. First, the core staff will produce stocks of virulent M. pulmonis organisms, which will be tested for use in all experiments to ensure a consistent supply with minimal variability. Second, the core will use standardized flow cytometric, DNA preparation and polymerase chain reaction procedures to genotype mice from the mutant colonies that will be used in the three projects. Third, the core will provide access to and assist in novel and newly developed techniques for real-time viewing of living cells in the ainways and lung through a collaboration with the UCSF Biological Imaging Development Center (www.ucsf edu/bidc/). Centralizing and coordinating infection, genotyping procedures, and imaging approaches will avoid duplication of resources, ensure that standardized procedures are used by all groups, facilitate the exchange of information, and foster collaborative interactions.