Tsarfaty I, Sandovsky-Losica H, Mittelman L, Berdicevsky I, Segal E
Department of Human Microbiology, Sackler School of Medicine, Tel Aviv University, Tel Aviv, Israel.
FEMS Microbiol Lett. 2000 Aug 15;189(2):225-32. doi: 10.1111/j.1574-6968.2000.tb09235.x.
Attachment of Candida albicans, an important opportunistic pathogen, to host tissues is an initial step in the development of the infection. The events occurring in the fungal and in the host cells after interaction are poorly understood. In this study we concentrated on the events occurring in the mammalian cells after the interaction with Candida, with emphasis on the cytoskeleton actin. Human cell line cells (HEp2) were exposed to C. albicans or C. albicans-secreted material (culture filtrate) (actin-rearranging Candida-secreted factor, arcsf). The HEp2 cells were examined for cellular changes using confocal laser microscopy (CLSM), transmission and scanning electron microscopy (TEM and SEM). The CLSM studies, using fluorescein isothiocyanate-labeled C. albicans and rhodamine phalloidin actin staining, revealed yeasts adhering to the HEp2 cells or internalized into the cells, with actin surrounding the fungi. Furthermore, actin rearrangement from filamentous network to actin aggregates was noticed. Interaction between the HEp2 cells and C. albicans could be demonstrated also by SEM and TEM after a 2-4-h exposure of the cells to the fungus. Yeasts and hyphae were found attaching to the surface and within the cells. CLSM studies revealed that exposure of HEp2 cells to arcsf was also followed by cellular actin rearrangement, reduced membrane ruffling and decreased cellular motility. The effect was dose- and time-dependent. All these data indicate that the interaction of Candida with HEp2 cells involves signaling events and affects the cellular actin.
白色念珠菌是一种重要的机会致病菌,其附着于宿主组织是感染发展的初始步骤。真菌与宿主细胞相互作用后发生的事件目前尚不清楚。在本研究中,我们专注于哺乳动物细胞与念珠菌相互作用后发生的事件,重点研究细胞骨架肌动蛋白。将人细胞系(HEp2)细胞暴露于白色念珠菌或白色念珠菌分泌的物质(培养滤液)(肌动蛋白重排念珠菌分泌因子,arcsf)。使用共聚焦激光显微镜(CLSM)、透射电子显微镜和扫描电子显微镜(TEM和SEM)检查HEp2细胞的细胞变化。CLSM研究使用异硫氰酸荧光素标记的白色念珠菌和罗丹明鬼笔环肽肌动蛋白染色,结果显示酵母附着于HEp2细胞或内化入细胞,肌动蛋白围绕真菌。此外,还观察到肌动蛋白从丝状网络重排为肌动蛋白聚集体。在将细胞暴露于真菌2 - 4小时后,SEM和TEM也证实了HEp2细胞与白色念珠菌之间的相互作用。发现酵母和菌丝附着于细胞表面及细胞内。CLSM研究表明,将HEp2细胞暴露于arcsf后也会发生细胞肌动蛋白重排、膜皱褶减少和细胞运动性降低。这种效应具有剂量和时间依赖性。所有这些数据表明,念珠菌与HEp2细胞的相互作用涉及信号事件并影响细胞肌动蛋白。