Mills S D, Finlay B B
Biotechnology Laboratory, University of British Columbia, Vancouver, Canada.
Microb Pathog. 1994 Dec;17(6):409-23. doi: 10.1006/mpat.1994.1086.
Invasion of the cultured epithelial cell lines HeLa, Henle-407, and Caco-2 (polarized and nonpolarized) by Salmonella typhi and Salmonella typhimurium were compared using conventional gentamicin invasion assays. Additionally, the mechanisms of invasion and intracellular trafficking by S. typhi and S. typhimurium were compared in HeLa cells using indirect immunofluorescence microscopy. S. typhi strain Ty2 was invasive in all human cell lines tested, including apical uptake into polarized Caco-2 cell monolayers. This strain also replicated at levels similar to S. typhimurium strain SL1344 inside HeLa and Henle-407 cells. Indirect immunofluorescence microscopy confirmed that S. typhi, like S. typhimurium, induced membrane ruffles and cytoskeletal rearrangements upon contact with HeLa cell surfaces. Ruffling induced by S. typhi and S. typhimurium was accompanied by macropinocytosis of the fluid phase endocytic marker fluorescein-dextran-sulphate and by aggregation of cell surface class I MHC heavy chain. Intracellular lysosomal trafficking of S. typhi and S. typhimurium in HeLa cells was also studied. The lysosomal membrane glycoprotein marker h-lamp-2 colocalized with S. typhi-containing vacuoles, as previously shown for S. typhimurium. The soluble lysosomal enzyme marker cathepsin D also was found within S. typhi-containing vacuoles to the same extent as previously published for S. typhimurium. The results from this study suggest that S. typhi and S. typhimurium use similar mechanisms for invasion and intracellular trafficking in cultured human epithelial cells.
使用传统的庆大霉素侵袭试验,比较了伤寒沙门氏菌和鼠伤寒沙门氏菌对培养的上皮细胞系HeLa、Henle - 407和Caco - 2(极化和非极化)的侵袭情况。此外,使用间接免疫荧光显微镜,在HeLa细胞中比较了伤寒沙门氏菌和鼠伤寒沙门氏菌的侵袭机制和细胞内运输情况。伤寒沙门氏菌Ty2菌株在所有测试的人类细胞系中均具有侵袭性,包括对极化的Caco - 2细胞单层的顶端摄取。该菌株在HeLa和Henle - 407细胞内的复制水平与鼠伤寒沙门氏菌SL1344菌株相似。间接免疫荧光显微镜证实,伤寒沙门氏菌与鼠伤寒沙门氏菌一样,在与HeLa细胞表面接触时会诱导膜褶皱和细胞骨架重排。伤寒沙门氏菌和鼠伤寒沙门氏菌诱导的褶皱伴随着液相内吞标记物荧光素 - 葡聚糖 - 硫酸盐的巨胞饮作用以及细胞表面I类MHC重链的聚集。还研究了HeLa细胞中伤寒沙门氏菌和鼠伤寒沙门氏菌的细胞内溶酶体运输。溶酶体膜糖蛋白标记物h - lamp - 2与含有伤寒沙门氏菌的液泡共定位,这与之前对鼠伤寒沙门氏菌的研究结果一致。可溶性溶酶体酶标记物组织蛋白酶D在含有伤寒沙门氏菌的液泡中的发现程度与之前报道的鼠伤寒沙门氏菌相同。这项研究的结果表明,伤寒沙门氏菌和鼠伤寒沙门氏菌在培养的人类上皮细胞中使用相似的侵袭和细胞内运输机制。