Molecular mechanisms of caveolin-1 controlled infection of endothelial cells by Influenza A and Ebola viruses

这项拟议的项目涉及埃博拉病毒和甲型流感病毒引起的病毒性出血热(VHF)中导致内皮细胞紊乱的分子机制。讨论了三个病原体决定因素。首先，我们将使用严格亲内皮的流感禽疫病毒来研究病毒诱导的内皮结构和屏障功能的直接紊乱。其次，我们将阐明重组埃博拉病毒可溶性糖蛋白(SGP、GP1和Delta肽)是否干扰内皮细胞。第三，我们将详细揭示埃博拉病毒和马尔堡病毒感染的单核/巨噬细胞释放的宿主细胞因子对内皮屏障功能的影响。利用功能(剪切力、阻抗谱)生化、形态和分子生物学技术，我们引起了人们对细胞间连接、焦点接触和细胞骨架蛋白的关注。在细胞系和内皮细胞中表达的荧光融合蛋白将被用来跟踪特定宿主细胞蛋白在不同病原体决定因素影响下的命运。这些研究将导致识别导致内皮功能紊乱的蛋白质和机制，并可能有助于设计治疗药物和开发VHF的预防策略。

The proposed project deals with the molecular mechanisms leading to disturbances of endothelial cells in viral hemorrhagic fever (VHF) caused by Ebola and influenza A viruses. Three pathogen determinants are addressed. Firstly, we will use the influenza fowl plague virus, which is strictly endothelial tropic to investigate direct virus induced disturbances of endothelial structure and barrier function. Secondly, we will clarify whether or not recombinant soluble glycoproteins (sGP, GP1 and delta peptide) of Ebola viruses disturb endothelial cells. Thirdly, we will uncover in detail the influence of host cell factors that are released from Ebola virus and Marburg virus infected monocytes/macrophages on endothelial barrier function. Using functional (shear stress, impedance spectroscopy) biochemical, morphological and molecular biological techniques we draw attention to proteins of the intercellular junctions, focal contacts and cytoskeleton. Fluorescent fusion proteins expressed in cell lines and endothelial cells will be used to follow the fate of particular host cell proteins under the influence of the different pathogen determinants. The investigations will lead to identification of the proteins and mechanisms responsible for endothelial disturbances and might help to design therapeutic agents and the development of preventative strategies in VHF.