Measuring pH, ROS Production, Maturation, and Degradation in Dendritic Cell Phagosomes Using Cytofluorometry-Based Assays
Measuring pH, ROS Production, Maturation, and Degradation in Dendritic Cell Phagosomes Using Cytofluorometry-Based Assays
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DOI：
10.1007/978-1-60761-421-0_25
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发表时间：
2010-01-01
期刊：
影响因子：
--
通讯作者：
Amigorena, Sebastian
中科院分区：
文献类型：
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作者：
Savina, Ariel;Vargas, Pablo;Amigorena, Sebastian
吞噬体是一种复杂的细胞器，在被病原体、垂死细胞或细胞碎片的吞噬细胞摄取后形成。吞噬体首先与内体，然后与溶酶体的高度动态相互作用导致吞噬体成熟为吞噬溶酶体。与其他吞噬细胞相反，它将摄入的颗粒降解为氨基酸，树突状细胞仅部分降解摄入的蛋白质，保留短肽用于适应性免疫应答的开始。我们已经修改了一系列的乳胶珠为基础的技术，以前报道，为了分析吞噬体成熟使用流式细胞术。基于与被吞噬颗粒结合的特异性探针的命运的技术，对吞噬体pH、降解或氧化遗迹进行分析。这些技术是非常敏感和定量的。
Phagosomes arc complex organelles that form after ingestion by phagocytic cells of pathogens, dying cells, or cell debris. Highly dynamic interactions of phagosomes first with endosomes and then with lysosomes lead to the maturation of phagosomes into phagolysosomes. Contrary to other phagocytes, which degrade ingested particles to amino acids, dendritic cells only partially degrade ingested proteins, preserving short peptides for the onset of adaptive immune responses. We have modified a series of latex bead-based techniques, previously reported, in order to analyze phagosome maturation Using flow cytometry. The analysis of the phagosomal pH, degradation, or oxidation relics on techniques based on the fate of specific probes bound to particles to be phagocytosed. These techniques arc very sensitive and quantitative.