Imaging Liver Immunobiology In Vivo in Real-time

实时体内肝脏免疫生物学成像

• 批准号: 6871311
• 负责人: Ian NICHOLAS Crispe
• 金额: $ 19.69万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-03-15 至 2006-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6871311
• 关键词: CD95 molecule; Kupffer' s cell; adeno associated virus group; antigens; apoptosis; bioimaging /biomedical imaging; cell cell interaction; cytotoxic T lymphocyte; genetically modified animals; immune response; laboratory mouse; leukocyte activation /transformation; liver cells; liver imaging /visualization /scanning; luciferin monooxygenase; lymphatic tissue; nuclear factor kappa beta; technology /technique development; transfection /expression vector

DESCRIPTION (provided by applicant): This proposal aims to develop Luciferase-based imaging to enhance and extend studies of the interaction of CD8+ T cells with the liver, and in particular the intra-hepatic apoptosis of CD8+ T cells. These studies derive from a parent project, entitled "Peripheral T Cell Deletion in Liver" (5RO1-AI37554-09). To correctly interpret the mechanism of CD8+ T cell accumulation and apoptosis in the liver, it is essential to know where the T cells first encounter antigen. In Specific Aim 1, we will use a luciferase reporter that is linked to the IL-2 promoter, and thus expressed in activated T cells, to determine whether T cell activation is seen first in lymphoid tissue, or first in the liver. We will examine this using both an experimental model driven by exogenous peptide, and a model driven by an Adeno-Associated Virus Vector that is optimized to infect hepatocytes. The interaction of CD8+ T cells with the liver appears to involve direct recognition of antigen, and we hypothesize that aspects of this recognition involve Kupffer Cells. Therefore, in Specific Aim 2 we will use a transgenic Luciferase reporter for NF-kappaB to determine whether CD8+ T cells accumulating in liver cause Kupffer Cell activation. When activated CD8+ T cells are trapped in the liver at the end of an immune response, there is damage to hepatocytes, which we term "collateral damage" since it does not appear to involve direct CTL function against hepatocytes. Several lines of evidence suggest that Fas and FasL are involved in such damage, but it is not clear on which cells the FasL is expressed, nor whether the FasL expression is closely followed by hepatocyte death. Thus in Aim 3 we will use a transgenic Luciferase reporter driven by the FasL promoter to test when and in which cells F as L is expressed during collateral damage. The studies proposed are consistent with the exploratory/developmental nature of the R21mechanism, since we aim to use a newly-available imaging technology to study T cell accumulation, T cell activation, and the effect of T cells on the liver of mice. This method can be used to study individual experimental mice longitudinally, thus refining experimentation by reducing the total number of mice required to obtain data.

描述（由申请人提供）：该提案旨在开发基于荧光素酶的成像，以增强和扩展 CD8+ T 细胞与肝脏相互作用的研究，特别是 CD8+ T 细胞的肝内凋亡。这些研究源自一个名为“肝脏中的外周 T 细胞删除”(5RO1-AI37554-09) 的父项目。为了正确解释 CD8+ T 细胞在肝脏中积累和凋亡的机制，有必要了解 T 细胞首先遇到抗原的位置。在具体目标 1 中，我们将使用与 IL-2 启动子连接并因此在激活的 T 细胞中表达的荧光素酶报告基因，以确定 T 细胞激活是首先在淋巴组织中出现，还是首先在肝脏中出现。我们将使用由外源肽驱动的实验模型和由经过优化以感染肝细胞的腺相关病毒载体驱动的模型来检查这一点。 CD8+ T 细胞与肝脏的相互作用似乎涉及对抗原的直接识别，我们假设这种识别的各个方面涉及库普弗细胞。因此，在具体目标 2 中，我们将使用 NF-kappaB 转基因荧光素酶报告基因来确定肝脏中积累的 CD8+ T 细胞是否会导致库普弗细胞激活。当激活的 CD8+ T 细胞在免疫反应结束时被困在肝脏中时，就会对肝细胞造成损害，我们将其称为“附带损害”，因为它似乎不涉及针对肝细胞的直接 CTL 功能。多项证据表明 Fas 和 FasL 参与了此类损伤，但尚不清楚 FasL 在哪些细胞上表达，也不清楚 FasL 表达是否紧随肝细胞死亡。因此，在目标 3 中，我们将使用由 FasL 启动子驱动的转基因荧光素酶报告基因来测试在附带损伤期间 F as L 何时以及在哪些细胞中表达。所提出的研究与 R21 机制的探索/发展性质一致，因为我们的目标是使用新的成像技术来研究 T 细胞积累、T 细胞激活以及 T 细胞对小鼠肝脏的影响。该方法可用于纵向研究个体实验小鼠，从而通过减少获取数据所需的小鼠总数来完善实验。