Institute of Innate Immunity, University Hospital Bonn, University of Bonn, Bonn, Germany.
Methods Mol Biol. 2022;2459:169-177. doi: 10.1007/978-1-0716-2144-8_16.
The detection of pathogen- or danger-associated molecular patterns during an inflammatory injury triggers the activation of cytosolic sensors known as inflammasomes. Once stimulated, these protein complexes can connect to the adaptor protein ASC, which in turn recruits the effector enzyme caspase-1, forming a polymeric structure known as ASC speck. This protein scaffold is responsible for processing cytokines of the IL-1 family into their active forms and evoking the cleavage of gasdermin D, ultimately leading to cell death by pyroptosis. Due to its micrometric size, the specks are used as a readout for inflammasome activation and for the better comprehension of this important immune pathway. In this chapter, a detailed protocol is presented for the study of the formation of inflammasome specks in living cells using confocal microscopy.
在炎症损伤过程中，病原体或危险相关分子模式的检测会触发细胞溶质传感器（称为炎性体）的激活。一旦受到刺激，这些蛋白复合物可以与衔接蛋白 ASC 连接，后者反过来招募效应酶半胱天冬酶-1，形成一种称为 ASC 斑点的聚合结构。这种蛋白质支架负责将白细胞介素-1 家族的细胞因子加工成其活性形式，并引发天冬氨酸半胱氨酸酶-1 的切割，最终通过细胞焦亡导致细胞死亡。由于其微小的尺寸，斑点被用作炎性体激活的读出器，并更好地理解这个重要的免疫途径。在本章中，提供了使用共聚焦显微镜研究活细胞中炎性体斑点形成的详细方案。
