Oishi Shinji, Takano Ryosuke, Tamura Satoshi, Tani Shinya, Iwaizumi Moriya, Hamaya Yasushi, Takagaki Kosuke, Nagata Toshi, Seto Shintaro, Horii Toshinobu, Osawa Satoshi, Furuta Takahisa, Miyajima Hiroaki, Sugimoto Ken
First Department of Medicine, Hamamatsu University School of Medicine, Hamamatsu, Shizuoka, Japan.
Department of Health Science, Hamamatsu University School of Medicine, Hamamatsu, Shizuoka, Japan.
Immunology. 2016 Nov;149(3):320-328. doi: 10.1111/imm.12647. Epub 2016 Aug 9.
Bone-marrow-derived macrophages are divided into two phenotypically and functionally distinct subsets, M1 and M2 macrophages. Recently, it was shown that adoptive transfer of M2-polarized peritoneal macrophages reduced the severity of experimental colitis in mice. However, it is still unclear whether peritoneal macrophages possess the same ability to be polarized to cells with functionally different phenotypes and cytokine production patterns as bone-marrow-derived macrophages. To address this question, we examined the ability of peritoneal macrophages to be polarized to the M1 and M2 phenotypes and determined the specific cytokine profiles of cells with each phenotype. We showed that peritoneal macrophages, as well as bone-marrow-derived macrophages, were differentiated into M1 and M2 phenotypes following stimulation with interferon-γ (IFN-γ) and interleukin-4 (IL-4)/IL-13, respectively. Following in vitro stimulation with lipopolysaccharide, M2-polarized peritoneal macrophages predominantly expressed T helper type 2 (Th2) cytokines and regulatory cytokines, including IL-4, IL-13, transforming growth factor-β and IL-10, whereas M1-polarized peritoneal macrophages expressed negligible amounts of Th1 and pro-inflammatory cytokines. ELISA showed that M2-polarized peritoneal macrophages produced significantly more IL-10 than M1-polarized peritoneal macrophages. Notably, M2-polarized peritoneal macrophages contributed more to the suppression of T-cell proliferation than did M1-polarized peritoneal macrophages. The mRNA expression of Th2 cytokines, including IL-4 and IL-13, increased in T-cells co-cultured with M2-polarized macrophages. Hence, our findings showed that M2 polarization of peritoneal macrophages induced regulatory cytokine production and suppressed T-cell proliferation in vitro, and that resident peritoneal macrophages could be used as a new adoptive transfer therapy for autoimmune/inflammatory diseases after polarization to the regulatory phenotype ex vivo.
骨髓来源的巨噬细胞可分为两个表型和功能不同的亚群，即M1和M2巨噬细胞。最近的研究表明，过继转移M2极化的腹膜巨噬细胞可减轻小鼠实验性结肠炎的严重程度。然而，尚不清楚腹膜巨噬细胞是否具有与骨髓来源的巨噬细胞相同的极化能力，使其分化为具有功能不同的表型和细胞因子产生模式的细胞。为了解决这个问题，我们检测了腹膜巨噬细胞极化至M1和M2表型的能力，并确定了每种表型细胞的特定细胞因子谱。我们发现，腹膜巨噬细胞以及骨髓来源的巨噬细胞，分别在干扰素-γ（IFN-γ）和白细胞介素-4（IL-4）/IL-13刺激后分化为M1和M2表型。在用脂多糖进行体外刺激后，M2极化的腹膜巨噬细胞主要表达2型辅助性T细胞（Th2）细胞因子和调节性细胞因子，包括IL-4、IL-13、转化生长因子-β和IL-10，而M1极化的腹膜巨噬细胞表达的Th1和促炎细胞因子含量可忽略不计。酶联免疫吸附测定（ELISA）显示，M2极化的腹膜巨噬细胞产生的IL-10明显多于M1极化的腹膜巨噬细胞。值得注意的是，M2极化的腹膜巨噬细胞比M1极化的腹膜巨噬细胞对T细胞增殖的抑制作用更强。在与M2极化巨噬细胞共培养的T细胞中，包括IL-4和IL-13在内的Th2细胞因子的mRNA表达增加。因此，我们的研究结果表明，腹膜巨噬细胞的M2极化可诱导调节性细胞因子的产生并在体外抑制T细胞增殖，并且驻留的腹膜巨噬细胞在体外极化至调节表型后可作为自身免疫性/炎症性疾病的一种新的过继转移治疗方法。
