EOSINOPHIL ACTIVITIES IN MURINE MODELS OF LUNG DISEASE

小鼠肺病模型中的嗜酸性粒细胞活性

• 批准号: 6153471
• 负责人: JAMES Joseph LEE
• 金额: $ 45.75万
• 依托单位: MAYO CLINIC ARIZONA
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-30 至 2004-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6153471
• 关键词: bioassay; biomarker; biotechnology; diagnosis design /evaluation; enzyme linked immunosorbent assay; eosinophil; flow cytometry; gene targeting; genetically modified animals; high throughput technology; immunocytochemistry; immunofluorescence technique; inflammation; laboratory mouse; laboratory rabbit; laboratory rat; lung disorder; mass screening; method development; monoclonal antibody; noninvasive diagnosis; rapid diagnosis; respiratory hypersensitivity; western blottings

DESCRIPTION (Adapted from the applicant's abstract) The aims of this proposal are to provide the research community with high throughput phenotypic assays in the mouse to detect the presence of eosinophils and/or eosinophil-mediated activities. These objectives utilize unique transgenic and gene knockout animals to develop eosinophil-specific reagents, creating three independent strategies for screening large numbers of mice in studies of allergic inflammation such as pulmonary models of asthma: (I) rabbit polyclonal antisera for histological screens of paraffin sections; (ii) rat/mouse monoclonal antibodies for sensitive sandwich ELISA assays to quantify eosinophil activities (e.g., eosinophil degranulation in BAL samples); (iii) generation/identification of monoclonal antibodies against cell surface epitopes specific for "activated" eosinophils in FACS analyses of eosinophil tissue/organ recruitment (e.g., recruitment of activated eosinophils to the airway lumen in mouse models of pulmonary inflammation). In the short-term, these strategies represent independent endpoint assessments. However, in the context of a larger scientific community analyzing different murine model systems, the results from these strategies will generate benchmark parameters for high throughput evaluations of inflammatory responses. Thus, the reagents/methodologies developed in this proposal uniquely provide a multifaceted approach investigators will use in rapid screens of large numbers of animals. Moreover, by correlating these reproducible and quantitative assays of defined inflammatory markers with more complex phenotypic assessments, investigators will create indices with which detailed evaluation of mouse models are achieved in a quick cost-effective fashion. (End of Abstract.)

描述（改编自申请人摘要） 该提案的目的是为研究界提供高 在小鼠中进行通量表型测定以检测 嗜酸性粒细胞和/或嗜酸性粒细胞介导的活性。 这些目标利用 独特的转基因和基因敲除动物， 试剂，创造了三个独立的策略，用于筛选大量的 小鼠在过敏性炎症研究中，如哮喘的肺部模型： (I)用于石蜡切片组织学筛选的兔多克隆抗血清; (ii)大鼠/小鼠单克隆抗体，用于灵敏的夹心ELISA测定， 量化嗜酸性粒细胞活性（例如，BAL嗜酸性粒细胞脱颗粒 （iii）产生/鉴定抗 在流式细胞仪分析中对“活化的”嗜酸性粒细胞特异的细胞表面表位 嗜酸性粒细胞组织/器官募集（例如，招募活动 嗜酸性粒细胞向肺部炎症小鼠模型中的气道腔的转移）。 在短期内，这些策略代表了独立的终点 评估。 然而，在更大的科学界背景下， 分析不同的小鼠模型系统，这些策略的结果 将生成用于高通量评估的基准参数， 炎症反应。 因此，本发明中开发的试剂/方法学可用于本领域。 该提案独特地提供了一种多方面的方法，调查人员将使用这种方法， 快速筛选大量动物。 此外，通过将这些关联起来， 确定的炎症标志物的可重复和定量测定， 复杂的表型评估，研究人员将创建指数， 小鼠模型的详细评估是在一个快速的成本效益 时尚. (End抽象）。