National Medical Research Center for Obstetrics, Gynecology and Perinatology Named after Academician V.I. Kulakov of Ministry of Healthcare of Russian Federation, 117997 Moscow, Russia; Рeoples' Friendship University of Russia (RUDN University), 117198 Moscow, Russia.
National Medical Research Center for Obstetrics, Gynecology and Perinatology Named after Academician V.I. Kulakov of Ministry of Healthcare of Russian Federation, 117997 Moscow, Russia.
Biomed Pharmacother. 2021 Jul;139:111614. doi: 10.1016/j.biopha.2021.111614. Epub 2021 Apr 27.
Macrophages are a central component of innate immunity that play an important role in the defense of the organism. Macrophages are highly plastic and are activated by interaction with other cells and environmental factors. In this work, we study the effect of lipopolysaccharide on macrophages derived from the two most polar (CD14+ and CD16+ monocytes) as well as the intermediate subset of blood monocytes from healthy donors and assess what happens to the subset most prone to polarization on the transcriptomic and proteomic level. It has been shown that, according to primary pro-inflammatory polarization markers, their cytokine profile, and their phagocytic activity, macrophages derived from CD14+ monocytes exhibit higher sensitivity to inducers of pro-inflammatory polarization. Flow cytometry analysis revealed increased levels of CD86, while secretome analysis demonstrated significant increase of pro-inflammatory and anti-inflammatory cytokines observed in CD14+-derived macrophages, as compared to CD16+-derived macrophages in conditioned media. Assessment of the transcriptome and proteome of CD14+-derived macrophages with further bioinformatic analysis identified the most significant differences after polarization towards the pro-inflammatory phenotype. Immune-, membrane-, IFN-γ-, cytokine-, and defense-associated pathways were found significantly prevalent, while downregulated pathways were represented by RNA binding-, housekeeping-, exocytosis-, intracellular transport-, peptide and amide metabolic-related signaling. This data could be useful for macrophage-based cell therapeutics of cancer, as it provides additional background for the manipulation of donor monocytes intended for back transplantation.
巨噬细胞是先天免疫系统的核心组成部分，在机体防御中发挥着重要作用。巨噬细胞具有高度的可塑性，并通过与其他细胞和环境因素的相互作用而被激活。在这项工作中，我们研究了脂多糖对来自两个最极性（CD14+和 CD16+单核细胞）以及来自健康供体的血液中间单核细胞亚群的巨噬细胞的影响，并评估了最易极化的亚群在转录组和蛋白质组水平上发生的变化。已经表明，根据主要的促炎极化标志物、细胞因子谱和吞噬活性，来自 CD14+单核细胞的巨噬细胞对促炎极化诱导剂更敏感。流式细胞术分析显示 CD86 水平升高，而分泌组分析表明，与 CD16+衍生的巨噬细胞相比，在条件培养基中，CD14+衍生的巨噬细胞中观察到促炎和抗炎细胞因子显著增加。进一步的生物信息学分析表明，CD14+衍生的巨噬细胞在向促炎表型极化后，其转录组和蛋白质组的评估显示出最显著的差异。免疫、膜、IFN-γ、细胞因子和防御相关途径显著占优势，而下调途径则代表 RNA 结合、管家、胞吐、细胞内运输、肽和酰胺代谢相关信号。这些数据对于基于巨噬细胞的癌症细胞治疗可能是有用的，因为它为操纵旨在回输的供体单核细胞提供了额外的背景。
