EOSINOPHIL ACTIVITIES IN MURINE MODELS OF LUNG DISEASE

描述(改编自申请人的摘要) 这项建议的目的是为研究界提供高质量的 通过在小鼠中进行表型分析来检测是否存在 嗜酸性粒细胞和/或嗜酸性粒细胞介导的活动。这些目标利用 独特的转基因和基因敲除动物产生嗜酸性粒细胞特异性 试剂，创造了三个独立的策略来筛选大量的 过敏性炎症研究中的小鼠，如哮喘的肺模型： (I)用于石蜡切片组织学筛选的兔多克隆抗血清； (Ii)大鼠/小鼠的单抗，用于敏感的夹心ELISA法检测 定量嗜酸性粒细胞活动(例如，BAL中的嗜酸性粒细胞脱颗粒 样本)；(3)产生/鉴定抗 流式细胞仪分析“激活的”嗜酸性粒细胞的细胞表面表位 嗜酸性粒细胞组织/器官招募(例如，激活的 在小鼠肺部炎症模型中，嗜酸性粒细胞对气道腔的影响)。 在短期内，这些战略代表着独立的终点 评估。然而，在一个更大的科学界的背景下 分析不同的小鼠模型系统，这些策略的结果 将为高吞吐量评估生成基准参数 炎症反应。因此，在这方面开发的试剂/方法 提案独一无二地提供了调查人员将在 对大量动物进行快速筛选。此外，通过将这些关联起来 明确的炎症标志物的可重复性和定量分析 复杂的表型评估，调查人员将创建用于 对鼠标模型进行了详细的评估，实现了快速的性价比 时尚。(摘要结束。)

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.)