State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China.
PLoS One. 2011;6(11):e26544. doi: 10.1371/journal.pone.0026544. Epub 2011 Nov 3.
The interaction between mammalian host cells and bacteria is a dynamic process, and the underlying pathologic mechanisms are poorly characterized. Limited information describing the host-bacterial interaction is based mainly on studies using label-based endpoint assays that detect changes in cell behavior at a given time point, yielding incomplete information. In this paper, a novel, label-free, real-time cell-detection system based on electronic impedance sensor technology was adapted to dynamically monitor the entire process of intestinal epithelial cells response to Salmonella infection. Changes in cell morphology and attachment were quantitatively and continuously recorded following infection. The resulting impedance-based time-dependent cell response profiles (TCRPs) were compared to standard assays and showed good correlation and sensitivity. Biochemical assays further suggested that TCRPs were correlated with cytoskeleton-associated morphological dynamics, which can be largely attenuated by inhibitions of actin and microtubule polymerization. Collectively, our data indicate that cell-electrode impedance measurements not only provide a novel, real-time, label-free method for investigating bacterial infection but also help advance our understanding of host responses in a more physiological and continuous manner that is beyond the scope of current endpoint assays.
哺乳动物宿主细胞与细菌的相互作用是一个动态的过程，其潜在的病理机制尚未得到充分描述。基于标记终点测定的研究主要描述了宿主-细菌的相互作用，该方法可检测特定时间点细胞行为的变化，提供的信息并不完整。在本文中，我们采用了一种新颖的、无标记的、基于实时细胞检测系统的电子阻抗传感器技术，动态监测了肠道上皮细胞对沙门氏菌感染的反应全过程。感染后，细胞形态和黏附的变化被定量和连续地记录下来。基于阻抗的时间依赖性细胞反应谱（TCRPs）与标准测定方法进行了比较，显示出良好的相关性和灵敏度。生化测定进一步表明，TCRPs 与细胞骨架相关的形态动力学相关，肌动蛋白和微管聚合的抑制可显著降低 TCRPs。综上所述，我们的数据表明，细胞-电极阻抗测量不仅提供了一种新颖的、实时的、无标记的方法来研究细菌感染，而且有助于以更生理和连续的方式深入了解宿主反应，这是当前终点测定方法无法实现的。
