Immunogenomics and Inflammation Research Unit, Edouard Herriot Hospital, Hospices Civils de Lyon, Lyon, France.
University Claude Bernard Lyon 1, Lyon, France.
Front Immunol. 2019 Dec 20;10:2992. doi: 10.3389/fimmu.2019.02992. eCollection 2019.
Methotrexate (MTX) at low dose is a key drug for rheumatoid arthritis (RA). MTX is widely used alone or combined with biologics or steroids. The aim was to study its effects on cytokine production using an model with synoviocytes interacting with peripheral blood mononuclear cells (PBMC) to reproduce the interactions in RA synovium. Activated-PBMC were co-cultured with RA synoviocytes during 48 h. A dose-response of MTX was tested and different biotherapies (Infliximab, Tocilizumab, Abatacept and Rituximab) were added alone or in combination with MTX. Cytokine production (IL-17, IL-6, IL-1β, IFN-γ, and IL-10) was measured by ELISA. These results were compared with those obtained with steroids. MTX alone had a modest inhibitory effect on cytokine production compared to steroids. The most effective concentration was one of the lowest, 0.01 μg/ml, as for steroids. Infliximab was the most active biotherapy ( ≤ 0.05 for all cytokines) followed by Tocilizumab ( ≤ 0.05 for all cytokines except IL-6). Abatacept and Rituximab had a more restricted effect on cytokines ( ≤ 0.05 for IL-1β and IFN-γ). The combination MTX/biotherapies did not increase significantly the inhibition of cytokine production but some specific inhibitory effects were observed with Infliximab on IL-17 and IL-6, and with Abatacept and Rituximab on IL-1β. Low dose of MTX was at least as effective as high dose. The effects of the combination with biotherapies showed an important level of heterogeneity between the levels of some specific cytokines and the degree of inhibition with drugs.
甲氨蝶呤（MTX）低剂量是类风湿关节炎（RA）的关键药物。MTX 广泛单独使用或与生物制剂或类固醇联合使用。目的是使用滑膜细胞与外周血单核细胞（PBMC）相互作用的模型来研究其对细胞因子产生的影响，以模拟 RA 滑膜中的相互作用。在 48 小时内，将活化的 PBMC 与 RA 滑膜细胞共培养。测试了 MTX 的剂量反应，并单独或联合 MTX 添加了不同的生物疗法（英夫利昔单抗、托珠单抗、阿巴西普和利妥昔单抗）。通过 ELISA 测量细胞因子（IL-17、IL-6、IL-1β、IFN-γ 和 IL-10）的产生。将这些结果与类固醇获得的结果进行比较。与类固醇相比，MTX 单独对细胞因子产生的抑制作用较弱。最有效的浓度与类固醇一样，为最低浓度之一，即 0.01μg/ml。英夫利昔单抗是最有效的生物疗法（所有细胞因子均 ≤ 0.05），其次是托珠单抗（所有细胞因子均 ≤ 0.05，除了 IL-6）。阿巴西普和利妥昔单抗对细胞因子的作用更有限（IL-1β 和 IFN-γ 均 ≤ 0.05）。MTX/生物疗法的联合并未显著增加细胞因子产生的抑制作用，但观察到英夫利昔单抗对 IL-17 和 IL-6 以及阿巴西普和利妥昔单抗对 IL-1β 的抑制作用具有特定的抑制作用。低剂量 MTX 与高剂量一样有效。联合生物疗法的效果显示，某些特定细胞因子的水平和药物抑制程度之间存在重要的异质性。
