Eakin Amanda J, Ahmed Tahanver, McGeough Cathy M, Drain Stephen, Alexander H Denis, Wright Gary D, Gardiner Philip V, Small Dawn, Bjourson Anthony J, Gibson David S
Personalised Medicine Centre, School of Medicine, Ulster University, Altnagelvin Hospital, Londonderry BT47 6SB, UK.
The Patrick G Johnston Centre for Cancer Research, Queen's University Belfast, Belfast BT9 7AE, UK.
J Pers Med. 2022 Nov 9;12(11):1875. doi: 10.3390/jpm12111875.
Disease activity in rheumatoid arthritis (RA) is influenced by activation of circulating and synovial immune cells. Regulatory T cells (Tregs) and monocytes are key cells that drive inflammation in RA. This study investigated if a relationship exists between disease activity in RA and circulating Treg and monocyte numbers and phenotypes. A potential sialic acid (Sia) mediated link between Tregs and monocytes was also probed in vitro. Peripheral blood mononuclear cells (PBMCs) were isolated from RA patient (n = 62) and healthy control (n = 21) blood using density gradient separation. Flow cytometry was used to count and phenotype Treg and monocyte subsets, and to sort healthy control Tregs for Sia cell culture experiments. The effects of Sia on activated Treg FoxP3 and NFκB expression was assessed by flow cytometry and concentrations of secreted TNFα, IL-10 and IFNγ determined by ELISA. High disease activity RA patients who were unresponsive to disease modifying anti-rheumatic drugs (n = 31), have significantly lower relative numbers (percentages) of CD4+CD25+CD127− Tregs (p < 0.01) and memory CD45RA−FoxP3+ Tregs (p < 0.01), compared to low disease activity responders (n = 24). Relative numbers of non-classical CD169+ monocytes are associated with disease activity in RA (p = 0.012). Sia reduced Treg expression of FoxP3, NFκB and cytokines in vitro. A strong association has been identified between non-classical CD169+ monocytes and post-treatment disease activity in RA. This study also indicates that Sia can reduce Treg activation and cytokine release. We postulate that such a reduction could be mediated by interaction with sialyted proteins captured by CD169+ monocytes.
类风湿关节炎(RA)的疾病活动受循环和滑膜免疫细胞激活的影响。调节性T细胞(Tregs)和单核细胞是驱动RA炎症的关键细胞。本研究调查了RA疾病活动与循环Treg及单核细胞数量和表型之间是否存在关联。还在体外探究了Tregs与单核细胞之间潜在的唾液酸(Sia)介导的联系。使用密度梯度分离法从RA患者(n = 62)和健康对照者(n = 21)的血液中分离外周血单个核细胞(PBMCs)。采用流式细胞术对Treg和单核细胞亚群进行计数和表型分析,并分选健康对照者的Tregs用于Sia细胞培养实验。通过流式细胞术评估Sia对活化Treg的FoxP3和NFκB表达的影响,并通过酶联免疫吸附测定法测定分泌的TNFα、IL - 10和IFNγ的浓度。与低疾病活动反应者(n = 24)相比,对改善病情抗风湿药物无反应的高疾病活动RA患者(n = 31)的CD4 + CD25 + CD127− Tregs(p < 0.01)和记忆性CD45RA− FoxP3 + Tregs(p < 0.01)的相对数量(百分比)显著降低。非经典CD169 +单核细胞的相对数量与RA的疾病活动相关(p = 0.012)。Sia在体外降低了Treg的FoxP3、NFκB和细胞因子的表达。已确定非经典CD169 +单核细胞与RA治疗后疾病活动之间存在强关联。本研究还表明,Sia可降低Treg的活化和细胞因子释放。我们推测这种降低可能是通过与CD169 +单核细胞捕获的唾液酸化蛋白相互作用介导的。