Enhanced Local Lymph Node Assay Using Flow Cytometry

使用流式细胞术增强局部淋巴结检测

• 批准号: 6663758
• 负责人: George L. DeGeorge
• 金额: $ 54.67万
• 依托单位: MB RESEARCH LABORATORIES, INC.
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-30 至 2004-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6663758
• 关键词: alternatives to animals in research; biomarker; bromodeoxyuridine; contact dermatitis; flow cytometry; immunologic assay /test; immunotoxicity; laboratory mouse; lymph nodes; lymphocyte proliferation; technology /technique development

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.

描述（申请人的摘要）：尽管小鼠局部淋巴结测定 (LLNA) 可有效检测潜在的刺激物/敏化剂，该测定法， 以目前的标准形式：1）不能轻易区分某些类型 来自呼吸道和接触致敏物的刺激物； 2）利用适度 一定量的放射性物质； 3）需要增加动物数量 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品的反应。 为了加强和改进这项重要的测试，我公司实施了多项 对标准 LLNA 的创新修改。我们应用流式细胞术 该测定的技术可提高测定的灵敏度/特异性， 消除放射性物质的使用并减少动物数量 用于筛选。已由SBIR第一阶段的结果确定 研究表明，所提出的基于增强流式细胞术的 LLNA 在技术上和 商业上可行。然而，要将这种测定法开发成完全商业化的 我们公司将为化学、制药和 消费品行业，我们小组必须：1）验证增强版 LLNA 的； 2) 将基于细胞计数的 LLNA 建立为符合 GLP 的检测方法 我们公司和； 3）开发并提供各种版本的增强型LLNA 为我们的客户。