Schneider Katharina S, Thomas Christina J, Groß Olaf
Institut für Klinische Chemie und Pathobiochemie, Klinikum rechts der Isar, Technische Universität München, Munich, Germany.
Methods Mol Biol. 2013;1040:117-35. doi: 10.1007/978-1-62703-523-1_10.
Through its ability to control the proteolytic maturation and secretion of interleukin-1 family cytokines, the inflammasome occupies a central role in the activation of inflammation and also influences the shaping of adaptive immunity. Since it affects a multitude of different immune responses from autoinflammatory diseases to host defense, vaccine efficacy, and even cancer, it has become of interest to many researchers. Here, we describe a straightforward method for inflammasome assays in primary murine bone marrow--derived myeloid cells. The protocol encompasses cell handling, inflammasome activation and inhibition, as well as the detection of IL-1β, caspase-1, and IL-1α by ELISA and Western blot.
炎性小体通过其控制白细胞介素-1家族细胞因子的蛋白水解成熟和分泌的能力,在炎症激活中占据核心地位,并且还影响适应性免疫的形成。由于它影响从自身炎症性疾病到宿主防御、疫苗效力甚至癌症等多种不同的免疫反应,因此已引起许多研究人员的关注。在此,我们描述了一种用于原代小鼠骨髓来源的髓样细胞中炎性小体检测的简单方法。该方案包括细胞处理、炎性小体激活和抑制,以及通过酶联免疫吸附测定(ELISA)和蛋白质印迹法检测白细胞介素-1β(IL-1β)、半胱天冬酶-1(caspase-1)和白细胞介素-1α(IL-1α)。