Julla Jean-Baptiste, Ballaire Raphaelle, Ejlalmanesh Tina, Gautier Jean-François, Venteclef Nicolas, Alzaid Fawaz
Centre de Recherche des Cordeliers, INSERM, Sorbonne Université, USPC, Université Paris Descartes, Université Paris Diderot, Paris, France.
Department of Diabetes, Clinical Investigation Centre (CIC-9504), Assistance Publique-Hôpitaux de Paris, Lariboisière Hospital, Paris, France.
Methods Mol Biol. 2019;1951:33-48. doi: 10.1007/978-1-4939-9130-3_3.
Monocytes and macrophages are cells of the innate immune system, existing in circulation and permeating every tissue of the human body. These cells represent the first responders to stress, and their inflammatory activation forms part of virtually every human disease. It is for these reasons that several approaches have been developed in order to phenotypically and functionally analyze these cells and their subpopulations. These inflammatory cells have been consistently demonstrated to undergo inflammatory polarization through the engagement of lipid-activated nuclear receptors, namely, through PPARs and LXRs, which regulate both lipid metabolism and inflammation. Quantitative and phenotypic analyses of monocytes and macrophages are largely dependent on cytometric tools and antibody-based labelling of membrane markers, while functional analyses apply a range of in vitro approaches coupled with secondary analyses of gene or cytokine expression. An important aspect of phenotypic and functional analyses is the purification, enrichment, or appropriate differentiation of biological materials, ensuring experimentation is carried out on monocytes and/or macrophages and not on other cell types occupying the same physical niches. This chapter will focus on the most common analytical approaches applied to the mononuclear phagocytic system, namely, circulating monocytes and adipose tissue macrophages.
单核细胞和巨噬细胞是先天性免疫系统的细胞，存在于血液循环中，并渗透到人体的每个组织。这些细胞是应激的第一反应者，它们的炎症激活几乎是每种人类疾病的一部分。正是由于这些原因，已经开发了几种方法来对这些细胞及其亚群进行表型和功能分析。这些炎症细胞一直被证明通过脂质激活的核受体的参与而发生炎症极化，即通过PPARs和LXRs，它们调节脂质代谢和炎症。单核细胞和巨噬细胞的定量和表型分析在很大程度上依赖于细胞计数工具和基于抗体的膜标记物标记，而功能分析则应用一系列体外方法以及基因或细胞因子表达的二次分析。表型和功能分析的一个重要方面是生物材料的纯化、富集或适当分化，确保实验是在单核细胞和/或巨噬细胞上进行，而不是在占据相同物理微环境的其他细胞类型上进行。本章将重点介绍应用于单核吞噬系统的最常见分析方法，即循环单核细胞和脂肪组织巨噬细胞。
