BIOLOGICAL MODEL OF ENDOTHELIAL ACTIVATION

内皮激活的生物学模型

• 批准号: 6302185
• 负责人: LEE A SPORN
• 金额: $ 25.58万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-04-01 至 2001-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6302185
• 关键词: Rickettsia; Rickettsiales disease; apoptosis; bacteria infection mechanism; cellular pathology; cysteine endopeptidases; enzyme activity; free radical oxygen; human tissue; immunocytochemistry; nuclear factor kappa beta; p53 gene /protein; thrombosis; tissue /cell culture; vascular endothelium

The overall objective of this proposal is to gain further understanding of the role of the endothelial cell in disease by elucidating mechanisms involved in initiation and regulation of apoptosis. Endothelial cell apoptosis can occur in response to numerous environmental stimuli and has been implicated in several pathologic conditions. Infection of endothelial cells with the obligate intracellular bacterium, Rickettsia rickettsii, will be used as a model system. The endothelial cell is the primary target during human infection, which results in the disease known as Rocky Mountain spotted fever, and endothelial cell responses to intracellular R.rickettsii likely play an important role in the pathological events that characterize this disease. Intracellular infection with R.rickettsii likely play an important roles studies to transduce both a pro-and anti-apoptotic signal. Blocking activation of the transcription factor, nuclear factor-kappaB, during infection results in rapid host cell apoptosis. This provides a valuable pathophysiologically-relevant model system for study of the mechanisms involved in regulation of this cellular response. The studies described in Specific Aim 1 are designed to further characterize the R. rickettsii-induced apoptotic program, including study of kinetics, dependence on intracellular infection and critical aspects of the infection required for its initiation. Specific Aim 2 will explore involvement of known signal transduction pathways or events including generation of reactive oxygen species, caspase activation and p53. Specific Aim 3 will explore will explore the consequences of endothelial cell activation by R. rickettsii in the context of the naturally- occurring disease setting, wherein it will be determined if infection protects the endothelial cell from exogenous pro-apoptotic stimuli which may be generated by inflammation or immune cell recruitment Microscopic techniques will also be used to explore the correlation between intracellular infection, apoptosis infection and activation of nuclear factor-kappaB both in cell culture models and in an ex vivo model of infection. These studies will not only provide important insight into regulation of endothelial cell apoptosis, but also into the complex interplay of signaling events which occur as part of the host cell- parasite relationship.

这项建议的总体目标是进一步了解 通过阐明内皮细胞在疾病中的作用机制 参与细胞凋亡的启动和调控。内皮细胞 细胞凋亡可以发生在对多种环境刺激的反应中 与几种病理情况有牵连。感染性疾病 含胞内专性细菌立克次体的内皮细胞 Rickettsii，将被用作示范系统。血管内皮细胞是 在人类感染期间的主要靶点，这导致了疾病 被称为落基山斑点热，内皮细胞对 胞内立克次体可能在致病中起重要作用 以这种疾病为特征的病理事件。细胞内 立克次体感染可能在研究中发挥重要作用 转导促进和反对细胞凋亡的信号。阻止激活 核因子-kappaB在感染过程中的转录因子 导致宿主细胞迅速凋亡。这提供了一个有价值的 用于机制研究的病理生理学相关模型系统 参与了这种细胞反应的调节。这些研究描述了 在特定目标中，1被设计为进一步刻画R. 立克次体诱导的细胞凋亡程序，包括动力学研究， 对细胞内感染的依赖及其关键方面 引发疾病所需的感染。《特定目标2》将探索 已知的信号转导途径或事件的参与，包括 活性氧的产生、半胱氨酸天冬氨酸酶的激活和p53的表达。 特定目标3将探索内皮细胞的后果 里氏杆菌在自然-环境中激活细胞- 发生疾病设置，其中将确定是否感染 保护内皮细胞免受外源性促凋亡刺激 可能是由炎症或免疫细胞募集产生的微观 技术也将被用来探索两者之间的相关性 细胞内感染、细胞凋亡感染和核激活 因子-kappaB在细胞培养模型和体外模型中的表达 感染。这些研究不仅将为 调控内皮细胞的凋亡，也进入复合体 作为宿主细胞一部分发生的信号事件的相互作用- 寄生虫关系。