Makita Naoyuki, Hizukuri Yoshiyuki, Yamashiro Kyoko, Murakawa Masao, Hayashi Yasuhiro
Faculty of Exploratory Pharmacology, Asubio Pharma Co., Ltd., 6-4-3, Minatojima-Minamimachi, Chuo-ku, Kobe 650-0047, Japan.
Faculty of Exploratory Technology, Asubio Pharma Co., Ltd., 6-4-3, Minatojima-Minamimachi, Chuo-ku, Kobe 650-0047, Japan.
Int Immunol. 2015 Mar;27(3):131-41. doi: 10.1093/intimm/dxu090. Epub 2014 Sep 29.
M2 macrophages have been subdivided into subtypes such as IL-4-induced M2a and IL-10-induced M2c in vitro. Although it was reported that IL-10 stimulation leads to an increase in IL-4Rα, the effect of IL-4 and IL-10 in combination with macrophage subtype differentiation remains unclear. Thus, we sought to clarify whether IL-10 enhanced the M2 phenotype induced by IL-4. In this study, we showed that IL-10 enhanced IL-4Rα expression in M-CSF-induced bone marrow-derived macrophages (BMDMs). Global gene expression analysis of M2 macrophages induced by IL-4, IL-10 or IL-4 + IL-10 showed that IL-10 enhanced gene expression of M2a markers induced by IL-4 in M-CSF-induced BMDMs. Moreover, IL-4 and IL-10 synergistically induced CCL24 (Eotaxin-2) production. Enhanced CCL24 expression was also observed in GM-CSF-induced BMDMs and zymosan-elicited, thioglycolate-elicited and naive peritoneal macrophages. CCL24 is a CCR3 agonist and an eosinophil chemoattractant. In vitro, IL-4 + IL-10-stimulated macrophages produced a large amount of CCL24 and increased eosinophil migration, which was inhibited by anti-CCL24 antibody. We also showed that IL-4 + IL-10-stimulated (but not IL-4 or IL-10 alone) macrophages transferred into the peritoneum of C57BL/6J mice increased eosinophil infiltration into the peritoneal cavity. These results demonstrate that IL-4 + IL-10-simulated macrophages have enhanced M2a macrophage-related gene expression, CCL24 production and eosinophil infiltration-inducing activity, thereby suggesting their contribution to eosinophil-related diseases.
M2巨噬细胞在体外已被细分为多种亚型,如IL-4诱导的M2a和IL-10诱导的M2c。尽管有报道称IL-10刺激会导致IL-4Rα增加,但IL-4和IL-10联合对巨噬细胞亚型分化的影响仍不清楚。因此,我们试图阐明IL-10是否增强了IL-4诱导的M2表型。在本研究中,我们发现IL-10增强了M-CSF诱导的骨髓来源巨噬细胞(BMDM)中IL-4Rα的表达。对由IL-4、IL-10或IL-4 + IL-10诱导的M2巨噬细胞进行的全基因组表达分析表明,IL-10增强了M-CSF诱导的BMDM中IL-4诱导的M2a标志物的基因表达。此外,IL-4和IL-10协同诱导CCL24(嗜酸性粒细胞趋化因子-2)的产生。在GM-CSF诱导的BMDM以及酵母聚糖诱导的、巯基乙酸盐诱导的和天然腹膜巨噬细胞中也观察到CCL24表达增强。CCL24是一种CCR3激动剂和嗜酸性粒细胞趋化剂。在体外,IL-4 + IL-10刺激的巨噬细胞产生大量CCL24并增加嗜酸性粒细胞迁移,这被抗CCL24抗体抑制。我们还发现,转移到C57BL/6J小鼠腹膜中的IL-4 + IL-10刺激的巨噬细胞(而非单独的IL-4或IL-10刺激的巨噬细胞)增加了嗜酸性粒细胞向腹腔的浸润。这些结果表明,IL-4 + IL-10刺激的巨噬细胞具有增强的M2a巨噬细胞相关基因表达、CCL24产生和嗜酸性粒细胞浸润诱导活性,从而提示它们在嗜酸性粒细胞相关疾病中的作用。