Carta Tania, Razzuoli Elisabetta, Fruscione Floriana, Zinellu Susanna, Meloni Dionigia, Anfossi Antonio, Chessa Bernardo, Dei Giudici Silvia, Graham Simon P, Oggiano Annalisa, Franzoni Giulia
Department of Animal Health, Istituto Zooprofilattico Sperimentale della Sardegna, 07100 Sassari, Italy.
School of Veterinary Medicine, University of Sassari, 07100 Sassari, Italy.
Animals (Basel). 2021 Apr 12;11(4):1098. doi: 10.3390/ani11041098.
Macrophages are phagocytic cells involved in maintaining tissue homeostasis and defense against pathogens. Macrophages may be polarized into different functionally specialized subsets. M2c macrophages arise following stimulation with IL-10 or TGF-β and mediate anti-inflammatory and tissue repair functions. M2c macrophages remain poorly characterized in the pig, thus we investigated the impact of these regulatory cytokines on porcine monocyte-derived macrophages (moMΦ). The phenotype and functionality of these cells was characterized though confocal microscopy, flow cytometry, ELISA, and RT-qPCR. Both cytokines induced CD14 and MHC II DR down-regulation and reduced IL-6, TNF-α, and CD14 expression, suggestive of an anti-inflammatory phenotype. Interestingly, neither IL-10 or TGF-β were able to trigger IL-10 induction or release by moMΦ. Differences between these cytokines were observed: stimulation with IL-10, but not TGF-β, induced up-regulation of both CD16 and CD163 on moMΦ. In addition, IL-10 down-regulated expression of IL-1β and IL-12p40 4h post-stimulation and induced a stronger impairment of moMΦ ability to respond to either TLR2 or TLR4 agonists. Overall, our results provide an overview of porcine macrophage polarization by two immunosuppressive cytokines, revealing differences between IL-10 and TGF-β, and reporting some peculiarity of swine, which should be considered in translational studies.
巨噬细胞是参与维持组织稳态和抵御病原体的吞噬细胞。巨噬细胞可极化为不同功能特化的亚群。M2c巨噬细胞在白细胞介素-10(IL-10)或转化生长因子-β(TGF-β)刺激后产生,并介导抗炎和组织修复功能。猪的M2c巨噬细胞特征仍不清楚,因此我们研究了这些调节性细胞因子对猪单核细胞衍生巨噬细胞(moMΦ)的影响。通过共聚焦显微镜、流式细胞术、酶联免疫吸附测定(ELISA)和逆转录定量聚合酶链反应(RT-qPCR)对这些细胞的表型和功能进行了表征。两种细胞因子均诱导CD14和主要组织相容性复合体II类DR(MHC II DR)下调,并降低白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)和CD14表达,提示具有抗炎表型。有趣的是,IL-10和TGF-β均不能触发moMΦ诱导或释放IL-10。观察到这些细胞因子之间的差异:用IL-10刺激而非TGF-β刺激可诱导moMΦ上CD16和CD163上调。此外,IL-10在刺激后4小时下调白细胞介素-1β(IL-1β)和白细胞介素-12p40表达,并对moMΦ对Toll样受体2(TLR2)或Toll样受体4(TLR4)激动剂的反应能力造成更强损害。总体而言,我们的结果概述了两种免疫抑制细胞因子对猪巨噬细胞极化的影响,揭示了IL-10和TGF-β之间的差异,并报道了猪的一些特性,这些特性在转化研究中应予以考虑。