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.)
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