Karassek Sascha, Starost Laura, Solbach Johanna, Greune Lilo, Sano Yasuteru, Kanda Takashi, Kim KwangSik, Schmidt M Alexander
From the Institute of Infectiology, Center for Molecular Biology of Inflammation, Westfälische Wilhelms-Universität Münster, D-48149 Münster, Germany.
the Department of Neurology and Clinical Neuroscience, Yamaguchi University Graduate School of Medicine, Yamaguchi, Japan, and.
J Biol Chem. 2015 Oct 9;290(41):24835-43. doi: 10.1074/jbc.M115.650101. Epub 2015 Aug 31.
Pertussis toxin (PTx), an AB5 toxin and major virulence factor of the whooping cough-causing pathogen Bordetella pertussis, has been shown to affect the blood-brain barrier. Dysfunction of the blood-brain barrier may facilitate penetration of bacterial pathogens into the brain, such as Escherichia coli K1 (RS218). In this study, we investigated the influence of PTx on blood-brain barrier permissiveness to E. coli infection using human brain-derived endothelial HBMEC and TY10 cells as in vitro models. Our results indicate that PTx acts at several key points of host cell intracellular signaling pathways, which are also affected by E. coli K1 RS218 infection. Application of PTx increased the expression of the pathogen binding receptor gp96. Further, we found an activation of STAT3 and of the small GTPase Rac1, which have been described as being essential for bacterial invasion involving host cell actin cytoskeleton rearrangements at the bacterial entry site. In addition, we showed that PTx induces a remarkable relocation of VE-cadherin and β-catenin from intercellular junctions. The observed changes in host cell signaling molecules were accompanied by differences in intracellular calcium levels, which might act as a second messenger system for PTx. In summary, PTx not only facilitates invasion of E. coli K1 RS218 by activating essential signaling cascades; it also affects intercellular barriers to increase paracellular translocation.
百日咳毒素(PTx)是引起百日咳的病原体百日咳博德特氏菌的一种AB5毒素和主要毒力因子,已被证明会影响血脑屏障。血脑屏障功能障碍可能会促进细菌病原体进入大脑,如大肠杆菌K1(RS218)。在本研究中,我们使用人脑来源的内皮细胞HBMEC和TY10细胞作为体外模型,研究了PTx对血脑屏障对大肠杆菌感染的通透性的影响。我们的结果表明,PTx作用于宿主细胞内信号通路的几个关键点,这些点也受到大肠杆菌K1 RS218感染的影响。PTx的应用增加了病原体结合受体gp96的表达。此外,我们发现STAT3和小GTP酶Rac1被激活,它们被认为对于细菌入侵至关重要,细菌入侵涉及宿主细胞肌动蛋白细胞骨架在细菌进入位点的重排。此外,我们表明PTx诱导VE-钙黏蛋白和β-连环蛋白从细胞间连接显著重新定位。宿主细胞信号分子的观察到的变化伴随着细胞内钙水平的差异,这可能作为PTx的第二信使系统。总之,PTx不仅通过激活关键信号级联促进大肠杆菌K1 RS218的入侵;它还影响细胞间屏障以增加细胞旁转运。