The proinflammatory protease caspase-1 plays pivotal roles in central pathways of innate immunity, thereby contributing to pathogen clearance. Beside its physiological role, dysregulated activity of caspase-1 is known to contribute to an increasing number of diseases. In this study, we optimized and validated a low-volume human whole blood assay facilitating the measurement of caspase-1 activation and inflammasome-related gene expression upon stimulation of the NLRP3, NLRC4 or AIM2 inflammasome. Using the NLRP3 inflammasome specific inhibitor MCC950, we were able to measure the activity of canonical or alternative NLRP3 pathways, AIM2 and NLRC4 inflammasomes in whole blood. Based on our data we assume a superposition of NLRP3 and NLRC4 inflammasome activities in human whole blood following stimulation with S. typhimurium. The optimized whole blood assay may be suitable for diagnostic and research purposes for pediatric patients who can only donate small amounts of blood.

促炎性蛋白酶caspase-1在先天免疫的主要途径中起关键作用，从而有助于病原体清除。除了其生理作用外，caspase-1活性失调还导致了越来越多的疾病。在这项研究中，我们优化和验证了一种低容量的人全血检测方法，该方法可促进NLRP3，NLRC4或AIM2炎性小体的刺激，从而测量caspase-1激活和炎性小体相关的基因表达。使用NLRP3炎性体特异性抑制剂MCC950，我们能够测量全血中规范性或替代性NLRP3途径，AIM2和NLRC4炎性体的活性。根据我们的数据，我们假设鼠伤寒沙门氏菌刺激后人全血中的NLRP3和NLRC4炎性体活性叠加。。优化的全血分析可能适合仅捐赠少量血液的小儿患者的诊断和研究目的。