Institute of Hygiene and Microbiology, University of Würzburg, Josef-Schneider-Str. 2, Bldg. E1, 97080 Würzburg, Germany.
Infect Immun. 2010 Aug;78(8):3392-403. doi: 10.1128/IAI.00012-10. Epub 2010 Jun 7.
Staphylococcus aureus is a major cause of a variety of both local and systemic infections. It can invade human host cells, a process that may account for disseminated and recurrent infections. S. aureus postinvasion events in nonprofessional phagocytes are only partially understood. While morphological data suggest a phagosomal escape, there is a lack of corroborating functional data. Using a combination of pH determination and morphological techniques, we have tested the integrity of Staphylococcus-containing phagosomes in 293 (HEK-293), HeLa, and EA.hy926 cells over time. Rapid acidification of S. aureus-containing phagosomes occurred and was sustained for up to 24 h. All S. aureus strains tested displayed equally sustained intraphagosomal pH levels without exhibiting any correlation with pH level and hemolytic activity. The membrane morphology of the phagosomal compartment was heterogeneous, even under conditions where acidic pH was fully maintained, an observation incompatible with phagolysosomal membrane destruction. As an exception, S. aureus strain 6850 showed a reduced phagosomal acidification signal 6 h after invasion. Additionally, only strain 6850 failed to localize to LAMP-1-positive vesicles in HeLa cells, although this was observed only rarely. Several other strongly beta-hemolytic strains did not modulate phagolysosomal pH, suggesting that S. aureus alpha-toxin and beta-toxin are not sufficient for this process. Taken together, our data suggest that S. aureus-containing phagolysosomes generally remain functionally intact in nonprofessional phagocytes, thereby contrasting with transmission electron micrographic results.
金黄色葡萄球菌是多种局部和全身感染的主要原因。它可以侵入人体宿主细胞,这一过程可能导致播散性和复发性感染。非专业吞噬细胞中金黄色葡萄球菌入侵后的事件仅部分被理解。虽然形态学数据表明存在吞噬体逃逸,但缺乏确证的功能数据。我们使用 pH 测定和形态学技术的组合,随时间测试了 293(HEK-293)、HeLa 和 EA.hy926 细胞中含金黄色葡萄球菌的吞噬体的完整性。含金黄色葡萄球菌的吞噬体迅速酸化,并持续长达 24 小时。所有测试的金黄色葡萄球菌菌株均显示出同样持续的吞噬体内 pH 水平,与 pH 水平和溶血活性没有任何相关性。吞噬体隔室的膜形态是异质的,即使在完全维持酸性 pH 的条件下,这一观察结果与吞噬溶酶体膜破坏不兼容。作为一个例外,金黄色葡萄球菌菌株 6850 在入侵后 6 小时显示出降低的吞噬体酸化信号。此外,只有菌株 6850 未能在 HeLa 细胞中定位于 LAMP-1 阳性囊泡,尽管这种情况很少见。其他几种强烈的β溶血菌株不会调节吞噬溶酶体 pH,这表明金黄色葡萄球菌α-毒素和β-毒素不足以完成这一过程。总之,我们的数据表明,非专业吞噬细胞中的含金黄色葡萄球菌吞噬溶酶体通常保持功能完整,这与透射电子显微镜结果形成对比。