Pseudomonas aeruginosa N-3-oxo-dodecanoyl-homoserine Lactone Elicits Changes in Cell Volume, Morphology, and AQP9 Characteristics in Macrophages.
Pseudomonas aeruginosa N-3-oxo-dodecanoyl-homoserine Lactone Elicits Changes in Cell Volume, Morphology, and AQP9 Characteristics in Macrophages.
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DOI：
10.3389/fcimb.2016.00032
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发表时间：
2016
影响因子：
5.7
通讯作者：
Vikström E
中科院分区：
文献类型：
--
作者：
Holm A;Magnusson KE;Vikström E
群体感应(QS)通信使铜绿假单胞菌能够共同控制其种群密度、生物被膜和毒力因子的产生。QS信号分子，如N-3-氧代-十二酰基-L-高丝氨酸内酯(3O-C12-HSL)，也可以影响宿主细胞的行为，例如，通过调节人类白细胞的趋化、迁移和吞噬。此外，宿主水平衡和水通道水通道蛋白(AQP)对细胞的形态和功能至关重要，因为AQP与细胞骨架和信号级联反应是间接的。在这里，我们研究了铜绿假单胞菌3O-C12-HSL如何影响细胞形态、面积、体积以及AQP9在人原代巨噬细胞中的表达和分布，方法包括定量聚合酶链式反应、免疫印迹、二维和三维实时成像、共聚焦成像和纳米级成像。因此，3O-C12-HSL增加了巨噬细胞的细胞体积和面积，并诱导了巨噬细胞的细胞形状和突起波动，这一过程暂时是由水通过AQP9(巨噬细胞中的主要AQP)通过细胞膜的通量驱动的。此外，3O-C12-HSL还上调了AQP9在蛋白和mRNA水平的表达。伴随着全细胞AQP9荧光强度的增强和AQP9重新分布到前导和拖尾区，同时伴随着巨噬细胞内细胞面积的增加。最后，纳米成像提供了AQP9在3O-C12-HSL刺激的细胞板脂区域内的动力学和结构的细节。我们认为，铜绿假单胞菌和巨噬细胞相互作用中的这些新事件可能会影响天然免疫细胞对抗细菌的有效性，从而解决感染和炎症的早期阶段。
Quorum sensing (QS) communication allows Pseudomonas aeruginosa to collectively control its population density and the production of biofilms and virulence factors. QS signal molecules, like N-3-oxo-dodecanoyl-L-homoserine lactone (3O-C12-HSL), can also affect the behavior of host cells, e.g., by modulating the chemotaxis, migration, and phagocytosis of human leukocytes. Moreover, host water homeostasis and water channels aquaporins (AQP) are critical for cell morphology and functions as AQP interact indirectly with the cell cytoskeleton and signaling cascades. Here, we investigated how P. aeruginosa 3O-C12-HSL affects cell morphology, area, volume and AQP9 expression and distribution in human primary macrophages, using quantitative PCR, immunoblotting, two- and three-dimensional live imaging, confocal and nanoscale imaging. Thus, 3O-C12-HSL enhanced cell volume and area and induced cell shape and protrusion fluctuations in macrophages, processes tentatively driven by fluxes of water across cell membrane through AQP9, the predominant AQP in macrophages. Moreover, 3O-C12-HSL upregulated the expression of AQP9 at both the protein and mRNA levels. This was accompanied with enhanced whole cell AQP9 fluorescent intensity and redistribution of AQP9 to the leading and trailing regions, in parallel with increased cell area in the macrophages. Finally, nanoscopy imaging provided details on AQP9 dynamics and architecture within the lamellipodial area of 3O-C12-HSL-stimulated cells. We suggest that these novel events in the interaction between P. aeruginosa and macrophage may have an impact on the effectiveness of innate immune cells to fight bacteria, and thereby resolve the early stages of infections and inflammations.