Enhanced Local Lymph Node Assay Using Flow Cytometry

描述(申请人？S摘要)：虽然小鼠局部淋巴结检 (LLNA)在检测潜在的刺激物/敏感剂方面是有效的，这种测试， 在其当前的标准形式中：1)不能容易地区分某些类型的 来自呼吸道和接触敏感剂的刺激物；2)使用适中的 放射性物质的数量和；3)需要增加动物的数量 确定不同的终点(即增殖、免疫表型) 充分描述动物对局部使用的化学物质的反应。 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进行了创新修改。我们已经应用了流式细胞术 该检测技术以提高检测的灵敏度/特异性， 消除放射性物质的使用，减少动物的数量 用于筛选。这是由SBIR第一阶段的结果确定的 研究表明，拟议的基于流式细胞术的增强LLNA在技术上和 商业上是可行的。然而，为了将这种检测方法完全商业化 潜力，我们公司将提供给化学品、制药和 消费品行业，我们集团必须：1)验证增强版 2)建立基于细胞学的LLNA作为符合GLP的检测方法 我公司和；3)开发和提供各种版本的增强型LLNA 为我们的客户服务。

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.