Among human macrophages polarised to different phenotypes, the M-CSF-oriented cells present the highest pro-inflammatory response to the rheumatoid arthritis-specific immune complexes containing ACPA
Among human macrophages polarised to different phenotypes, the M-CSF-oriented cells present the highest pro-inflammatory response to the rheumatoid arthritis-specific immune complexes containing ACPA
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DOI：
10.1136/annrheumdis-2015-208887
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发表时间：
2016-12-01
影响因子：
27.4
通讯作者：
Sebbag, Mireille
中科院分区：
文献类型：
--
作者：
Clavel, Cyril;Ceccato, Laurie;Sebbag, Mireille
目的在类风湿关节炎(RA)患者炎症滑膜中，抗瓜氨酸蛋白(ACPA)自身抗体可能在瓜氨酸纤维蛋白沉积上形成免疫复合体(IC)。我们发现，在体外，这种ACPA-IC激活了M-CSF分化的巨噬细胞的促炎细胞因子反应。我们的目的是评估巨噬细胞极化对这一反应的影响。方法将CD14阳性的单核细胞培养在M-CSF、干扰素-γ、IL-4或IL-10的存在下。用流式细胞术分析极化巨噬细胞特异性标志物的表达。用体外产生的抗瓜氨酸蛋白免疫复合体的自身抗体(ACPA-IC)刺激其分泌细胞因子，并在培养上清液中进行检测。结果干扰素-γ极化的细胞表达高水平的CD64和CD80。CD14的低表达和CD206的高表达是IL-4极化细胞的特征。IL-10或M-CSF可上调CD14、CD32和CD163的表达。两种细胞均缺乏CD80，CD64、CD200R和CD206的表达相似。对ACPA-IC的反应，干扰素-γ、IL-4和IL-10对IL-1β、IL-6和IL-8的分泌作用相似。而后者的IL-1ra：IL-1β比值最高，肿瘤坏死因子(TNF)-α：IL-10比值最低。相反，M-CSF暴露的巨噬细胞分泌最高水平的促炎细胞因子，表现出高的TNF-α：IL-10比率和最低的IL-1ra：IL-1β比率。结论尽管IL-10极化的巨噬细胞和M-CSF极化的巨噬细胞对ACPA-IC的细胞因子反应明显不同，但它们的表型相似。M-CSF极化细胞表现出最高的促炎潜能。由于M-CSF在RA滑膜中含量丰富，可能促使巨噬细胞对局部形成的ACPA-IC产生强烈的促炎细胞因子反应。
Objectives In the inflamed synovium of patients with rheumatoid arthritis (RA), autoantibodies to citrullinated proteins (ACPA) probably form immune complexes (IC) on deposits of citrullinated fibrin. We showed that in vitro such ACPA-IC activate a pro-inflammatory cytokine response in M-CSF-differentiated macrophages. Our objective was to evaluate how macrophage polarisation influences this response.Methods CD14-positive monocytes from healthy donors were cultured in the presence of M-CSF, IFN-gamma, interleukin (IL)-4 or IL-10. Expression of markers specific for polarised macrophages was analysed by flow cytometry. Their cytokine secretion was prompted by in vitro generated autoantibodies to citrullinated proteins immune complexes (ACPA-IC) and assayed in the culture supernatants.Results IFN-gamma-polarised cells exhibited high levels of CD64 and CD80. Low expression of CD14 and high expression of CD206 characterised the IL-4-polarised cells. Exposure to IL-10 or M-CSF raised the expression of CD14, CD32 and CD163. The two cell types lacked CD80 and exhibited similar expression of CD64, CD200R and CD206. In response to ACPA-IC, the secretion of IL-1 beta, IL-6 and IL-8 was similar among cells exposed to IFN-gamma, IL-4 or IL-10. However, the later cells were associated with the highest IL-1Ra: IL-1 beta ratio and the lowest tumour necrosis factor (TNF)-alpha:IL-10 ratio. Conversely, M-CSF-exposed cells secreted the highest levels of pro-inflammatory cytokines, exhibited a high TNF-alpha:IL-10 ratio and the lowest IL-1Ra:IL-1 beta ratio.Conclusions Despite their phenotypic similarity, IL-10-polarised and M-CSF-polarised macrophages clearly differ in their cytokine response to ACPA-IC. M-CSF-polarised cells exhibit the highest pro-inflammatory potential. Since M-CSF is abundant in the RA synovium, therein it probably drives macrophages towards a strong pro-inflammatory cytokine response to the locally formed ACPA-IC.