Enhanced Local Lymph Node Assay Using Flow Cytometry

描述（申请人？虽然小鼠局部淋巴结试验 （LLNA）可有效检测潜在刺激物/致敏物，该试验， 在其目前的标准形式中：1）不能容易地区分某些类型的 来自呼吸道和接触致敏物的刺激物; 2）使用中度 （1）放射性物质;（2）需要更多的动物 为了确定不同的端点（即，增殖，免疫表型）， 充分表征动物对局部施用的化学品的反应。 为了加强和改进这项重要的测试，我们公司实施了几项 对标准LLNA进行了创新性修改。我们应用流式细胞仪 该测定的技术以增加测定的灵敏度/特异性， 消除放射性物质的使用，减少动物的数量 用于筛选。它是由SBIR第一阶段的结果确定的 研究表明，拟议的增强型基于流式细胞术的LLNA在技术上和 商业上可行。然而，为了将该测定开发到其完全商业化， 潜力，我们公司将提供给化学，制药和 消费品行业，我们的团队必须：1）验证增强版 2）将基于细胞计数的LLNA确立为符合GLP的试验， 我们公司和; 3）开发和提供各种版本的增强LLNA 为我们的客户。

DESCRIPTION (Applicant?s abstract): Although the murine Local Lymph Node Assay (LLNA) is effective at detecting potential irritants/sensitizers, this assay, in its current standard form: 1) cannot readily differentiate some types of irritants from respiratory and contact sensitizers; 2) utilizes moderate amounts of radioactive material and; 3) requires increased numbers of animals to determine the different endpoints (i.e., proliferation, immunophenotype) to fully characterize the response of an animal to a topically-applied chemical. To enhance and improve this important test, our company has implemented several innovative modifications to the standard LLNA. We have applied flow cytometric techniques to this assay to increase the sensitivity/specificity of the assay, eliminate the use of radioactive material and decrease the number of animals used for screening. It has been determined by the results of SBIR Phase I studies that the proposed enhanced flow cytometry-based LLNA is technically and commercially feasible. However, to develop this assay to its full commercial potential, which our company would provide to the chemical, pharmaceutical and consumer products industries, our group must: 1) validate the enhanced version of the LLNA; 2) establish the cytometry-based LLNA as a GLP compliant assay at our company and; 3) develop and provide various versions of the enhanced LLNA for our clients.