Visualizing the Role of Myeloid Cells During CNS Autoimmunity

在这个项目中，我们将研究骨髓细胞在实验性自身免疫性脑脊髓炎（EAE），多发性硬化症的动物模型的启动功能。以下问题将遵循：1.在EAE初期，髓样细胞迁移到CNS的步骤是什么？2.介导它们穿越血脑屏障（BBB）的分子线索是什么？3.髓样细胞在打开BBB中起什么作用？4.哪种髓样细胞介导初始抗原呈递给自身攻击性致脑炎T细胞？该项目的方法重点在于将双光子激光扫描显微镜与功能/分子分析相结合。通过用荧光染料染色或通过使用遗传编码的荧光蛋白质使髓样细胞可见。将通过实时2-PM观察细胞在BBB和不同CNS环境和EAE阶段的迁移行为。在急性和慢性EAE模型中，骨髓细胞与自身侵袭性T细胞的相互作用的分析将使用将被引入T细胞的功能标志物进行。这些标记物将使得T细胞和髓样APC之间的结合以及效应T细胞的连续活化的可视化成为可能。

In this project we will study the function of myeloid cells during the initiation of experimental autoimmune encephalomyelitis (EAE), an animal model for multiple sclerosis. The following questions will be followed: 1. What are the migratory steps of myeloid cells into the CNS during incipient EAE? 2. What are the molecular cues that mediate their transmigration of the blood brain barrier (BBB)? 3. What role do myeloid cells play in opening the BBB? 4. Which myeloid cells mediate the initial antigen presentation to autoaggressive encephalitogenic T cells? The methodic focus of this project lies in the combination of 2-photon laser scanning microscopy with functional/molecular analyses. Myeloid cells will be made visible by staining them with fluorescence dyes or by use of genetically encoded fluorescence proteins. The migratory behavior of the cells at the BBB and in the different CNS milieus and EAE phases will be visualized by live 2-PM. The analyses of the interactions of myeloid cells with autoagressive T cells in acute and chronic EAE models will be performed using functional markers which will be introduced into the T cells. These markers will make possible the visualization of the binding between T cells and myeloid APCs and the consecutive activation of the effector T cells.