天青颗粒
脱颗粒
颗粒(地质)
髓过氧化物酶
细胞生物学
吞噬体
化学
先天免疫系统
NADPH氧化酶
生物化学
生物
活性氧
炎症
吞噬作用
免疫学
受体
古生物学
作者
Samia Bedouhène,Pham My-Chan Dang,Margarita Hurtado-Nédelec,Jamel El-Benna
出处
期刊:Methods in molecular biology
日期:2019-11-15
卷期号:: 215-222
被引量:16
标识
DOI:10.1007/978-1-0716-0154-9_16
摘要
Neutrophils play a pivotal role in innate immunity and in the inflammatory reactions. Upon activation, neutrophils release several toxic molecules directed against microbial pathogens into the phagosome. These molecules include reactive oxygen species (ROS), myeloperoxidase, glucosidases, proteases, and antibacterial peptides. In resting cells these proteins and the enzyme responsible for ROS production (NOX2) are stored inside or at the membranes of different granules called azurophil or primary, specific or secondary, gelatinase or tertiary, and the secretory vesicles. Each granule has a specific density, content, and markers. Myeloperoxidase (MPO) is the azurophil granule marker, and the neutrophil-gelatinase-associated lipocalin (NGAL) is the specific granule marker. After cell activation by different stimuli, granule contents are released into the phagosome or in the extracellular space through a process called degranulation. Also during this process, membrane granules fuse with the phagosome and plasma membrane allowing expression of new markers at the cell surface. The degranulation can be assessed by measuring either the release of different proteins by neutrophils or the expression of granule markers at the plasma membrane. In this chapter, we describe the techniques used to measure degranulation of azurophil and specific neutrophil granules using different approaches such as measurement of MPO enzymatic activity and detection of MPO and NGAL proteins by SDS-PAGE and Western blot.
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