Infection Program, Biomedicine Discovery Institute and Department of Biochemistry & Molecular Biology, Monash University, Clayton, VIC, Australia.
Methods Mol Biol. 2022;2523:239-252. doi: 10.1007/978-1-0716-2449-4_15.
The NLRP3 inflammasome senses the activity of pore-forming toxins secreted by Staphylococcus aureus. The bacterial toxins compromise plasma membrane integrity which activates the NLRP3 inflammasome to induce host pore-forming proteins and cellular suicide, termed pyroptosis. Host cell death rates are routinely determined at pre-defined time points and on whole cell populations. To capture the dynamic interactions between bacterial pore-forming toxins and host cell death factors, we have applied live-cell imaging techniques capable of analyzing single cell events in real time. Here, we describe methods using live-cell imaging to determine the host responses, such as plasma membrane integrity, mitochondrial health, and apoptotic caspases, towards pore-forming toxins.
NLRP3 炎性小体可感知金黄色葡萄球菌分泌的形成孔毒素的活性。细菌毒素会损害质膜完整性，从而激活 NLRP3 炎性小体，诱导宿主形成孔蛋白并发生细胞自杀，即细胞焦亡。通常在预定时间点和整个细胞群中测定宿主细胞死亡率。为了捕获细菌形成孔毒素与宿主细胞死亡因子之间的动态相互作用，我们应用了能够实时分析单细胞事件的活细胞成像技术。在这里，我们描述了使用活细胞成像来确定宿主对形成孔毒素的反应的方法，例如质膜完整性、线粒体健康和凋亡半胱天冬酶。
