流式细胞术评估 ASC 寡聚化。

Assessment of ASC Oligomerization by Flow Cytometry.

机构信息

Biomedical Research Institute of Murcia (IMIB), Clinical University Hospital Virgen de la Arrixaca, Murcia, Spain.

Department of Biochemistry and Molecular Biology B and Immunology, Faculty of Medicine, University of Murcia,, Murcia, Spain.

出版信息

Methods Mol Biol. 2022;2459:1-9. doi: 10.1007/978-1-0716-2144-8_1.

DOI:10.1007/978-1-0716-2144-8_1

PMID:35212949

Abstract

Inflammasomes are multiprotein complexes that critically control different aspects of innate and adaptive immunity. Upon activation, inflammasome proteins oligomerize forming scaffolds to nucleate the apoptosis-associated speck-like protein containing a CARD (ASC) in filaments that will finally result in large ASC oligomers that are commonly named as ASC specks. In this chapter, we present a method to monitor NLRP3 or pyrin inflammasome activation in human monocytes upon extracellular ATP or Clostridium difficile toxin B treatment, respectively, by detecting intracellular oligomers of ASC by flow cytometry. This method could be used to evaluate the degree of inflammasome activation in blood samples from patients suffering from different chronic inflammatory diseases.

摘要

炎症小体是一种多蛋白复合物，对先天免疫和适应性免疫的不同方面起着关键的调控作用。在被激活后，炎症小体蛋白寡聚化形成支架，以核衣壳含有 CARD（ASC）的丝状物中的凋亡相关斑点样蛋白（ASC），最终导致通常称为 ASC 斑点的 ASC 大寡聚物。在本章中，我们介绍了一种通过流式细胞术检测细胞内 ASC 寡聚体来监测人单核细胞在外源 ATP 或艰难梭菌毒素 B 处理下 NLRP3 或 pyrin 炎症小体激活的方法。该方法可用于评估患有不同慢性炎症性疾病的患者血液样本中炎症小体激活的程度。