19F MRI PROBES AND AUTOIMMUNE DISEASE

这项建议包括技术开发部分和基本的免疫生物学部分。本研究的主要目的是：（1）开发新的细胞内MRI试剂和相关的细胞标记方案，并测量细胞标记效率和寿命;（2）通过定量细胞毒性、增殖和免疫学分析对标记的免疫细胞（如树突状细胞（DC）和巨噬细胞）进行生物学表征;和（3）体内研究，其中将标记的免疫细胞注射到NOD小鼠中并使用极高场（11.7T）MRI成像。我们将可视化静脉接种后标记细胞的生物分布和局部注射到特定组织部位后的延时细胞迁移。我们将测试的假设，不同的DC亚群有不同的迁移模式在体内的基础上，他们的成熟状态。使用NOD小鼠，我们将测试的假设，即成熟的树突状细胞，以防止糖尿病的能力是依赖于，在某种程度上，对他们的迁移能力。该技术的长期应用是监测体内细胞治疗剂的运输。该研究项目具有高度的跨学科性，我们 在匹兹堡地区组建了一个多元化的研究团队，拥有体内磁共振显微镜（Ahrens），免疫学（Morel）和生物试剂设计（Ernst/Waggoner）的专业知识。

This proposal has a technology development component and a basic immunobiological component. It consists of three Specific Aims: (1) development of novel intracellular MRI agents and associated cell labeling protocols, and measurement of the cell labeling efficiency and lifetime; (2) biological characterizations via quantitative cytotoxicity, proliferation, and immunological assays of labeled immune cells, such as dendritic cells (DCs) and macrophages; and (3) in vivo studies where labeled immune cells will be injected into NOD mice and imaged using extremely-high field (11.7 T) MRI. We will visualize the biodistribution of the labeled cells following intravenous inoculation and the time-lapse cell migration following focal injection into specific tissue sites. We will test the hypothesis that different subsets of DCs have different migratory patterns in vivo based on their maturation state. Using the NOD mouse we will test the hypothesis that the ability of mature DCs to prevent diabetes is dependent, in part, on their migratory capacity. A long-term application of this technology is monitoring the trafficking of cellular therapeutics in vivo. The proposed research project is highly interdisciplinary, and we have assembled a diverse team of investigators in the Pittsburgh area with expertise in in vivo magnetic resonance microscopy (Ahrens), immunology (Morel) and biological reagent design (Ernst/Waggoner).