Vogel Daphne Y S, Glim Judith E, Stavenuiter Andrea W D, Breur Marjolein, Heijnen Priscilla, Amor Sandra, Dijkstra Christine D, Beelen Robert H J
Department of Molecular Cell Biology & Immunology, VU University Medical Center, Amsterdam, The Netherlands; Department of Pathology, VU University Medical Center, Amsterdam, The Netherlands.
Department of Molecular Cell Biology & Immunology, VU University Medical Center, Amsterdam, The Netherlands; Department of Plastic and Reconstructive Surgery, VU University Medical Center, Amsterdam, The Netherlands.
Immunobiology. 2014 Sep;219(9):695-703. doi: 10.1016/j.imbio.2014.05.002. Epub 2014 May 28.
Macrophages form a heterogeneous cell population displaying multiple functions, and can be polarized into pro- (M1) or anti-inflammatory (M2) macrophages, by environmental factors. Their activation status reflects a beneficial or detrimental role in various diseases. Currently several in vitro maturation and activation protocols are used to induce an M1 or M2 phenotype. Here, the impact of different maturation factors (NHS, M-CSF, or GM-CSF) and activation methods (IFN-γ/LPS, IL-4, dexamethason, IL-10) on the macrophage phenotype was determined. Regarding macrophage morphology, pro-inflammatory (M1) activation stimulated cell elongation, and anti-inflammatory (M2) activation induced a circular appearance. Activation with pro-inflammatory mediators led to increased CD40 and CD64 expression, whereas activation with anti-inflammatory factors resulted in increased levels of MR and CD163. Production of pro-inflammatory cytokines was induced by activation with IFN-γ/LPS, and TGF-β production was enhanced by the maturation factors M-CSF and GM-CSF. Our data demonstrate that macrophage marker expression and cytokine production in vitro is highly dependent on both maturation and activation methods. In vivo macrophage activation is far more complex, since a plethora of stimuli are present. Hence, defining the macrophage activation status ex vivo on a limited number of markers could be indecisive. From this study we conclude that maturation with M-CSF or GM-CSF induces a moderate anti- or pro-inflammatory state respectively, compared to maturation with NHS. CD40 and CD64 are the most distinctive makers for human M1 and CD163 and MR for M2 macrophage activation and therefore can be helpful in determining the activation status of human macrophages ex vivo.
巨噬细胞构成了一个具有多种功能的异质性细胞群体，并且可被环境因素极化为促炎（M1）或抗炎（M2）巨噬细胞。它们的激活状态反映了在各种疾病中所起的有益或有害作用。目前，有几种体外成熟和激活方案用于诱导M1或M2表型。在此，我们确定了不同成熟因子（NHS、M-CSF或GM-CSF）和激活方法（IFN-γ/LPS、IL-4、地塞米松、IL-10）对巨噬细胞表型的影响。关于巨噬细胞形态，促炎（M1）激活刺激细胞伸长，而抗炎（M2）激活诱导细胞呈圆形外观。用促炎介质激活导致CD40和CD64表达增加，而用抗炎因子激活则导致MR和CD163水平升高。用IFN-γ/LPS激活诱导促炎细胞因子的产生，而成熟因子M-CSF和GM-CSF增强TGF-β的产生。我们的数据表明，体外巨噬细胞标志物表达和细胞因子产生高度依赖于成熟和激活方法。体内巨噬细胞激活要复杂得多，因为存在大量刺激因素。因此，根据有限数量的标志物在体外确定巨噬细胞激活状态可能是不确定的。从这项研究中我们得出结论，与用NHS成熟相比，用M-CSF或GM-CSF成熟分别诱导适度的抗炎或促炎状态。CD40和CD64是人类M1最具特征性的标志物，而CD163和MR是M2巨噬细胞激活的标志物，因此有助于在体外确定人类巨噬细胞的激活状态。
