Vinet Adrien F, Descoteaux Albert
INRS-Institut Armand Frappier, Laval, QC, Canada.
Methods Mol Biol. 2009;531:329-46. doi: 10.1007/978-1-59745-396-7_20.
Phagocytosis is the process by which cells engulf and destroy large particles such as pathogens or apoptotic cells. In this way, macrophages play a pivotal role in the resolution of microbial infections. However, many microorganisms have evolved efficient strategies to preempt the weaponry of macrophages. A better understanding of the components engaged in the phagosome formation and maturation is necessary to devise novel approaches aimed at counteracting these microbial strategies. Recently, large-scale approaches have been used to improve our understanding of phagosome functional properties by the identification of hundreds of proteins and by studying each of them. Presently, purification of pathogen-containing phagosomes presents several technical challenges, whereas the use of latex beads to isolate phagosomes presents many advantages because this system can mimic host-pathogen interactions during phagocytosis. This system thus remains the best approach to advance our knowledge of phagosome biology, notably when used in conjunction with functional approaches. In this chapter, we outline an approach for the isolation of large-scale phagosome preparations with high degrees of purity.
吞噬作用是细胞吞噬并破坏诸如病原体或凋亡细胞等大颗粒的过程。通过这种方式，巨噬细胞在解决微生物感染中发挥着关键作用。然而，许多微生物已经进化出有效的策略来抢先应对巨噬细胞的武器装备。为了设计出对抗这些微生物策略的新方法，有必要更好地了解参与吞噬体形成和成熟的成分。最近，大规模方法已被用于通过鉴定数百种蛋白质并对它们逐一进行研究来增进我们对吞噬体功能特性的理解。目前，纯化含病原体的吞噬体存在若干技术挑战，而使用乳胶珠来分离吞噬体具有许多优点，因为该系统可以模拟吞噬过程中的宿主 - 病原体相互作用。因此，该系统仍然是推进我们对吞噬体生物学认识的最佳方法，特别是与功能方法结合使用时。在本章中，我们概述了一种用于大规模分离高纯度吞噬体制剂的方法。
