19 FMRI PROBES AND AUTOIMMUNE DISEASE

该提案包括技术开发部分和基本免疫生物学部分。它包括三个具体目标：(1)开发新型的细胞内MRI试剂和相关的细胞标记方案，并测量细胞的标记效率和寿命；(2)通过定量的细胞毒性、增殖和免疫学分析标记的免疫细胞，如树突状细胞(DC)和巨噬细胞，鉴定其生物学特性；(3)体内研究，将标记的免疫细胞注射到NOD小鼠，并使用极高场(11.7T)MRI进行成像。我们将观察静脉接种后标记细胞的生物分布，以及焦点注射到特定组织部位后延时细胞的迁移。我们将根据DC的成熟状态来检验这一假设，即不同的DC亚群在体内有不同的迁移模式。利用NOD小鼠，我们将检验这一假设，即成熟DC预防糖尿病的能力部分依赖于它们的迁移能力。这项技术的长期应用是监测细胞治疗药物在体内的运输。拟议的研究项目是高度跨学科的，我们 已经在匹兹堡地区组建了一支多样化的研究团队，他们拥有体内磁共振显微镜(Ahrens)、免疫学(Morel)和生物试剂设计(Ernst/Waggoner)方面的专业知识。

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).