Immunology Section, Department Experimental Medical Science, Lund University, Lund, Sweden.
Truly Labs, Medicon Village, Lund, Sweden.
Methods Mol Biol. 2023;2674:261-282. doi: 10.1007/978-1-0716-3243-7_18.
Inflammasomes are large multiprotein complexes that assemble mainly in innate immune cells after detection of microbial or sterile insults. Activation of inflammasomes is a key proinflammatory event during infection, and many pathogens have evolved specific evasion mechanisms to evade or inhibit inflammasome activation. One such pathogen is the common bacterium group A Streptococcus (GAS), which causes a wide range of diseases of varying severity. GAS secretes a multitude of virulence factors whereof the pore-forming protein streptolysin O (SLO) is the main inflammasome activation determinant. Here we provide a protocol for reliable evaluation of inflammasome activation in murine bone marrow-derived macrophages (BMDM) infected with GAS, including instructions for generating BMDMs and growing the bacterium. This protocol can easily be modified to other bacterial pathogens, or human macrophages.
炎症小体是一种大型多蛋白复合物，主要在固有免疫细胞检测到微生物或无菌损伤后组装。炎症小体的激活是感染过程中关键的促炎事件，许多病原体已经进化出特定的逃避机制来逃避或抑制炎症小体的激活。一种这样的病原体是常见的 A 组链球菌（GAS），它会引起各种严重程度不同的疾病。GAS 分泌多种毒力因子，其中形成孔的蛋白链球菌溶血素 O（SLO）是主要的炎症小体激活决定因素。在这里，我们提供了一种使用 GAS 感染的鼠骨髓来源的巨噬细胞（BMDM）可靠评估炎症小体激活的方案，包括生成 BMDM 和培养细菌的说明。该方案可以很容易地修改为其他细菌病原体或人巨噬细胞。
