The Role of Calcium Ions in Lymphocyte Activation

描述（由申请人提供）：我们的实验室率先使用 膜片钳和光学技术来研究离子通道的作用， (Ca)T淋巴细胞中的信号传导。T细胞启动免疫反应， 与致敏抗原呈递细胞直接接触后的外源抗原 (APC)例如树突细胞。T细胞受体（TCR）接合导致免疫应答。 导致Ca依赖性基因表达的细胞溶质Ca（Ca）信号，细胞因子 分泌、增殖和分化。我们提出的实验 使用活细胞成像的方法将平行于这一系列事件。 双光子显微镜最近已经能够可视化的T和B细胞深 在完整的淋巴结内，并揭示了运动的动态模式， 细胞-细胞相互作用，提供了一个独特的机会，在体内成像 免疫系统功能。因此，我们的做法将旨在巩固以下方面的成果： 在抗原特异性原代T细胞上进行的体外和体内实验 从转基因小鼠和T细胞系。各种引进和遗传 编码的荧光报告基因将直接监测TCR（Ca）基因， 表达和刺激前后T细胞的分泌反应。一 本项目的进一步目标是比较 光环境下静止、活化和慢性活化的自身反应性T细胞 不同的离子通道表型。

DESCRIPTION (provided by applicant):Our laboratory has pioneered the use of patch-clamp and optical techniques to investigate the role of ion channels and (Ca) signaling in T lymphocytes. T cells initiate the immune response to foreign antigens following direct contact with primed antigen presenting cells (APC) such as dendritic cells. T cell receptor (TCR) engagement results in a cytosolic Ca (Ca) signal that leads to Ca dependent gene expression, cytokine secretion, proliferation, and differentiation. Our proposed experimental approach using live-cell imaging will parallel this sequence of events. Two-photon microscopy has recently enable visualization of T and B cells deep within intact lymph node, and has revealed a dynamic pattern of motility and cell-cell interaction, offering a unique opportunity for in vivo imaging of immune system function. Thus, our approach will aim to consolidate results from in vitro and in vivo experiments performed on antigen-specific primary T cells from transgenic mice and T cell lines. A variety of introduced and genetically encoded fluorescent reporters will monitor the TCR directly, (Ca), gene expression, and secretory responses in T cells before and after stimulation. A further goal of this project is to compare the functional responses of quiescent, activated, and chronically activated auto reactive T cells in light of their differing ion channel phenotypes.