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Real-Time Sensing of Enteropathogenic E. coli-Induced Effects on Epithelial Host Cell Height Cell-Substrate Interactions and Endocytic Processes by Infrared Surface Plasmon Spectroscopy

机译:实时检测肠致病性大肠杆菌诱导的上皮宿主细胞高度细胞-基质相互作用和内吞过程的红外表面等离子体光谱

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摘要

Enteropathogenic Escherichia coli (EPEC) is an important, generally non-invasive, bacterial pathogen that causes diarrhea in humans. The microbe infects mainly the enterocytes of the small intestine. Here we have applied our newly developed infrared surface plasmon resonance (IR-SPR) spectroscopy approach to study how EPEC infection affects epithelial host cells. The IR-SPR experiments showed that EPEC infection results in a robust reduction in the refractive index of the infected cells. Assisted by confocal and total internal reflection microscopy, we discovered that the microbe dilates the intercellular gaps and induces the appearance of fluid-phase-filled pinocytic vesicles in the lower basolateral regions of the host epithelial cells. Partial cell detachment from the underlying substratum was also observed. Finally, the waveguide mode observed by our IR-SPR analyses showed that EPEC infection decreases the host cell's height to some extent. Together, these observations reveal novel impacts of the pathogen on the host cell architecture and endocytic functions. We suggest that these changes may induce the infiltration of a watery environment into the host cell, and potentially lead to failure of the epithelium barrier functions. Our findings also indicate the great potential of the label-free IR-SPR approach to study the dynamics of host-pathogen interactions with high spatiotemporal sensitivity.
机译:肠致病性大肠杆菌(EPEC)是一种重要的,通常为非侵入性的细菌性病原体,可引起人类腹泻。微生物主要感染小肠的肠上皮细胞。在这里,我们应用了我们最新开发的红外表面等离子体共振(IR-SPR)光谱方法来研究EPEC感染如何影响上皮宿主细胞。 IR-SPR实验表明,EPEC感染导致被感染细胞的折射率强烈降低。在共聚焦和全内反射显微镜的协助下,我们发现该微生物扩大了细胞间隙,并诱导了宿主上皮细胞下部基底外侧区域中的液相填充的胞囊小泡的出现。还观察到部分细胞从下面的基底脱离。最后,通过我们的IR-SPR分析观察到的波导模式表明EPEC感染在一定程度上降低了宿主细胞的高度。总之,这些观察结果揭示了病原体对宿主细胞结构和内吞功能的新影响。我们建议这些变化可能会诱导水性环境渗透到宿主细胞中,并可能导致上皮屏障功能的衰竭。我们的发现还表明,无标记IR-SPR方法具有很高的时空敏感性,可用于研究宿主-病原体相互作用的动力学。

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