Objective To investigate the impact of glucocorticoids (GCs) and anti-rheumatic drugs on the lymphocyte activation gene-3 (LAG-3) and on programmed cell death-1 (PD-1) expression on synovial and peripheral cells ex-vivo. Methods Synovial fluid mononuclear cells (SFMCs) from psoriatic arthritis (PsA, n = 26) and rheumatoid arthritis (RA, n = 13) patients, SFCs from osteoarthritis (OA, n = 5) patients and peripheral blood mononuclear cells (PBMCs) of healthy donors (n = 14) were co-cultured with GCs, glucocorticoid receptor antagonist RU486, methotrexate (MTX) and biologics. LAG-3 and PD-1 expressions on immune subsets were analyzed by flow cytometry. Results GCs in PsA inhibited SFMCs growth vs medium (2.3 ± 0.4X105 vs 5.3 ± 0.7X105, respectively, p < 0.01) and markedly upregulated CD14+LAG-3+ cells (11.7 ± 2.4% vs 0.8 ± 0.3%, p < 0.0001, respectively), but not CD3+LAG-3+ and CD14+PD-1+ cells. MTX had no effect on CD14+LAG-3+ cells (0.7 ± 0.3%). The TNFi inhibitors, infliximab (IFX) and etanercept, but not IL-12/23i, upregulated CD14+LAG-3+ cells vs medium (2.0 ± 0.6% and 1.6 ± 0.4% vs 0.5 ± 0.1%, p < 0.03, respectively). SFMCs growth inhibition in both PsA and RA correlated with CD14+LAG-3+ cell upregulation (r = 0.53, p = 0.03). RU486 inhibited GC-induced CD14+LAG-3+ cell up-regulation in a dose-dependent manner compared with GC alone (5µM 5.3 ± 1.2% and 50µM 1.3 ± 0.5% vs 7.0 ± 1.4%, p < 0.003), but had no significant effect on CD14+LAG-3+ cells co-cultured with IFX. GCs in healthy donors’ PBMCs upregulated the immune subsets CD3+LAG-3+, CD14+LAG-3+ and CD14+PD-1+ cells. Conclusion This study proposes a novel regulatory mechanism of GCs and of TNFi mediated by LAG-3 upregulation in synovial monocytes and PBMCs. LAG-3 modulation may be a promising target for development of novel therapies for inflammatory arthritis.

中文翻译：

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糖皮质激素和肿瘤坏死因子抑制剂通过淋巴细胞激活基因 3 上调发挥滑膜炎症的新抑制途径：一项离体研究


目的 探讨糖皮质激素（GC）和抗风湿药物对离体滑膜和外周细胞淋巴细胞活化基因 3（LAG-3）和程序性细胞死亡 1（PD-1）表达的影响。方法 来自银屑病关节炎 (PsA, n = 26) 和类风湿性关节炎 (RA, n = 13) 患者的滑液单核细胞 (SFMC)、来自骨关节炎 (OA, n = 5) 患者的滑液单核细胞 (SFMC) 以及来自以下患者的外周血单核细胞 (PBMC)健康供体 (n = 14) 与 GC、糖皮质激素受体拮抗剂 RU486、甲氨蝶呤 (MTX) 和生物制剂共培养。通过流式细胞术分析免疫亚群上的 LAG-3 和 PD-1 表达。结果 PsA 中的 GC 相对于培养基抑制 SFMC 生长（分别为 2.3 ± 0.4X105 与 5.3 ± 0.7X105，p < 0.01），并显着上调 CD14+LAG-3+ 细胞（11.7 ± 2.4% 与 0.8 ± 0.3%，p %） 3C 0.0001，分别），但不是 CD3+LAG-3+ 和 CD14+PD-1+ 细胞。 MTX 对 CD14+LAG-3+ 细胞没有影响 (0.7 ± 0.3%)。 TNFi 抑制剂英夫利昔单抗 (IFX) 和依那西普，但不包括 IL-12/23i，与培养基相比，上调 CD14+LAG-3+ 细胞（2.0 ± 0.6% 和 1.6 ± 0.4% vs 0.5 ± 0.1%，p < 0.03，分别）。 PsA 和 RA 中的 SFMC 生长抑制与 CD14+LAG-3+ 细胞上调相关 (r = 0.53，p = 0.03)。与单独使用 GC 相比，RU486 以剂量依赖性方式抑制 GC 诱导的 CD14+LAG-3+ 细胞上调（5μM 5.3 ± 1.2% 和 50μM 1.3 ± 0.5% vs 7.0 ± 1.4%，p < 0.003），但对与IFX共培养的CD14+LAG-3+细胞无显着影响。健康捐献者 PBMC 中的 GC 上调免疫亚群 CD3+LAG-3+、CD14+LAG-3+ 和 CD14+PD-1+ 细胞。结论 本研究提出了滑膜单核细胞和 PBMC 中 LAG-3 上调介导的 GC 和 TNFi 的新调节机制。 LAG-3 调节可能是开发炎症性关节炎新疗法的一个有希望的目标。