BIOLOGICAL MODEL OF ENDOTHELIAL ACTIVATION

内皮激活的生物学模型

• 批准号: 6109727
• 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: 1999
• 资助国家: 美国
• 起止时间: 1999-04-01 至 2000-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6109727
• 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.

本建议的总体目标是进一步了解 内皮细胞在疾病中的作用， 参与细胞凋亡的启动和调节。内皮细胞 细胞凋亡可响应于多种环境刺激而发生， 与多种病理状况有关。感染 内皮细胞与专性胞内细菌立克次体 立克次氏体，将被用作模型系统。内皮细胞是 在人类感染过程中的主要目标，导致疾病 称为落基山斑疹热，以及内皮细胞对 胞内立克次氏体可能在 这种疾病的病理特征。细胞内 立克次体感染可能在研究中发挥重要作用， 表达促凋亡和抗凋亡信号。阻断激活 转录因子，核因子-κ B，在感染过程中， 导致宿主细胞快速凋亡。这提供了有价值的 病理生理学相关的模型系统，用于研究 参与调节这种细胞反应。描述的研究 在具体目标1中，设计用于进一步表征R. 立克次氏体诱导的凋亡程序，包括动力学研究， 对细胞内感染的依赖性和 感染是其启动所必需的。第2章将探索 涉及已知的信号转导途径或事件，包括 活性氧的产生、半胱天冬酶活化和p53。 具体目标3将探索内皮细胞 细胞激活R.自然环境中的立克次氏体 发生疾病的设置，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， 可能由炎症或免疫细胞募集产生 技术也将被用来探索之间的相关性， 细胞内感染、凋亡感染和核活化 在细胞培养模型和体外模型中， 感染这些研究不仅将提供重要的洞察力， 调节内皮细胞凋亡，也是复杂的 作为宿主细胞的一部分发生的信号事件的相互作用- 寄生虫关系