Haslinger Bettina, Strangfeld Katrin, Peters Georg, Schulze-Osthoff Klaus, Sinha Bhanu
Institute of Medical Microbiology, Medical School of University Hospital of Münster, Domagkstrasse 10, D-48149 Münster, Germany.
Cell Microbiol. 2003 Oct;5(10):729-41. doi: 10.1046/j.1462-5822.2003.00317.x.
Staphylococcus aureus infections can result in septic and toxic shock with depletion of immune cells and massive cytokine production. Recently, we showed that, in S. aureus-infected Jurkat T cells, alpha-toxin is the major mediator of caspase activation and apoptosis. Here, we investigated the mechanisms of cell death induced by alpha-toxin in peripheral blood mononuclear cells (MNC). We show that alpha-toxin is required and sufficient for S. aureus-induced cell death not only in transformed Jurkat T cells but also in MNC. Low alpha-toxin doses (3-30 ng ml-1) dose- and time-dependently induced apoptosis in both cell types, which was completely blocked by the caspase inhibitor zVAD-fmk. In Jurkat T cells and MNC, alpha-toxin induced the breakdown of the mitochondrial membrane potential and the intrinsic activation of caspase-3, -8 and -9. Interestingly, unlike in Jurkat T cells, apoptosis in MNC was additionally mediated by a caspase-9-independent component. MNC, but not Jurkat T cells, produced tumour necrosis factor (TNF)-alpha upon alpha-toxin stimulation. Blocking endogenous TNF-alpha with a TNF-alpha receptor antagonist partially decreased apoptosis in MNC. Our data therefore suggest that, whereas in Jurkat T cells apoptosis is solely mediated by the mitochondrial pathway, in MNC endogenous TNF-alpha and a death receptor-dependent pathway are also involved, which may contribute to depletion of immune cells during S. aureus infection.
金黄色葡萄球菌感染可导致败血症和中毒性休克,伴有免疫细胞耗竭和大量细胞因子产生。最近,我们发现,在感染金黄色葡萄球菌的Jurkat T细胞中,α-毒素是半胱天冬酶激活和细胞凋亡的主要介质。在此,我们研究了α-毒素在外周血单个核细胞(MNC)中诱导细胞死亡的机制。我们发现,α-毒素不仅在转化的Jurkat T细胞中,而且在MNC中,都是金黄色葡萄球菌诱导细胞死亡所必需且充分的。低剂量的α-毒素(3 - 30 ng/ml)在两种细胞类型中均呈剂量和时间依赖性地诱导细胞凋亡,而半胱天冬酶抑制剂zVAD - fmk可完全阻断这种凋亡。在Jurkat T细胞和MNC中,α-毒素诱导线粒体膜电位的破坏以及半胱天冬酶-3、-8和-9的内源性激活。有趣的是,与Jurkat T细胞不同,MNC中的细胞凋亡还由一个不依赖半胱天冬酶-9的成分介导。α-毒素刺激后,MNC而非Jurkat T细胞产生肿瘤坏死因子(TNF)-α。用TNF-α受体拮抗剂阻断内源性TNF-α可部分降低MNC中的细胞凋亡。因此,我们的数据表明,在Jurkat T细胞中,细胞凋亡仅由线粒体途径介导,而在MNC中,内源性TNF-α和死亡受体依赖性途径也参与其中,这可能导致金黄色葡萄球菌感染期间免疫细胞的耗竭。