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Detection of LC3‐Associated Phagocytosis (LAP)

机译：检测LC3相关吞噬作用（LAP）

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摘要

Phagocytes, notably macrophages, are critical sentinels of their environment, patrolling for and eradicating unwanted components. The ability of cells to process extracellular cargo in an appropriate manner is important for both clearance of the cargoand eventual return to homeostasis. Although the evolutionarily conserved pathway of autophagy involves the degradation and recycling of unnecessary or dysfunctional cellular components during starvation, we now appreciate that the reach of autophagy extends beyond nutrient deprivation, notably including cellular quality control (e.g., mitophagy) and host defense against internalized pathogens (i.e., xenophagy). Despite being seemingly disparate, autophagic functions are unified as conserved mechanismsfor containment and immunosuppression, suggesting an original immune function for autophagy. A recently described pathway called LC3‐associated phagocytosis (LAP) marries the ancient concepts of phagocytosis and autophagy, revealing new ways in which the autophagy machinery, in a molecularly distinct pathway, contributes to the inflammatory response. In this article, protocols to detect LAP by electron microscopy, immunofluorescence, flow cytometry, and phagosome purification are described, allowing the user to detect multiple characteristics of LAP in both qualitative and quantitative manners.
机译：吞噬细胞，特别是巨噬细胞，是他们环境的关键哨兵，巡逻和消除不需要的组件。以适当的方式对细胞来处理细胞外货物的能力对于船舶的清除，最终恢复到稳态。虽然自噬的进化保守途径涉及在饥饿期间对不必要或功能失调的细胞组分的降解和再循环，但我们现在欣赏自噬的覆盖范围超越营养剥夺，特别是包括细胞质量控制（例如，MITOCHAGY）和对抗内化病原体的宿主防御（即Xenophagy）。尽管看似不同，自噬功能统一作为储蓄和免疫抑制的保守机制，表明自噬的原始免疫功能。最近描述的途径称为LC3-相关吞噬作用（LAP）结合了吞噬作用和自噬的古代概念，揭示了自噬机械在分子明显的途径中有助于炎症反应的新方法。在本文中，描述了通过电子显微镜，免疫荧光，流式细胞术和吞噬物体检测液圈的协议，允许使用者以定性和定量方式检测膝盖的多个特征。

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  • 来源
    《Current Protocols in Cell Biology》 |2020年第1期|共20页
  • 作者

    Jennifer Martinez;

  • 作者单位

    Immunity Inflammation and Disease Laboratory National Institute of Environmental Health Sciences National Institutes of Health Research Triangle Park North Carolina;

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 细胞生物学;
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