Inhibition of cytokine production by methotrexate. Studies in healthy volunteers and patients with rheumatoid arthritis
Inhibition of cytokine production by methotrexate. Studies in healthy volunteers and patients with rheumatoid arthritis
复制标题
DOI：
10.1093/rheumatology/keg323
复制
发表时间：
2003-10-01
期刊：
影响因子：
5.5
通讯作者：
Aarden, LA
中科院分区：
文献类型：
--
作者：
Gerards, AH;de Lathouder, S;Aarden, LA
目标。分析甲氨蝶呤对类风湿关节炎(RA)的有益作用是否与抑制炎性细胞因子的产生有关。用健康志愿者和类风湿关节炎患者的全血(WB)和单个核细胞(MNC)研究了细胞因子的产生。用低聚脂糖(LOS)或金黄色葡萄球菌Cowan I(SAC)等细菌产物刺激单核细胞，或用抗CD3和CD28的单抗诱导多克隆T细胞活化。我们分析了甲氨蝶呤对这些系统中细胞因子产生的影响。我们发现甲氨蝶呤能抑制T细胞活化诱导的细胞因子的产生。被抑制的细胞因子包括白介素4(IL-4)、白介素13(IL-13)、干扰素-γ(IFN-γ)、肿瘤坏死因子-α(TNF-α)和粒-巨噬细胞集落刺激因子。在服用该药的RA患者的血浆中，可以看到很容易达到的抑制浓度。甲氨蝶呤对IL-8的产生几乎没有影响。此外，甲氨蝶呤对LOS或SAC诱导的IL-6、IL-8、IL-1β和肿瘤坏死因子α的产生仅有轻微的抑制作用。加入叶酸或胸腺嘧啶核苷和次黄嘌呤可逆转甲氨蝶呤对细胞因子产生的抑制作用。抑制所需的甲氨蝶呤浓度因供体而异。RA患者口服甲氨蝶呤10 mg后，2小时后抽血中细胞因子的产生受到明显抑制。事实证明，甲氨蝶呤是一种有效的抑制新鲜抽血中T细胞活化诱导的细胞因子产生的药物。这是由于抑制了嘌呤和嘧啶的从头合成。单核细胞产生的细胞因子几乎不受甲氨蝶呤的影响。
Objectives. To analyse whether the beneficial effects of methotrexate in rheumatoid arthritis (RA) could be due to inhibition of inflammatory cytokine production.Methods. Cytokine production was studied using whole blood (WB) and mononuclear cells (MNC) of healthy volunteers and RA patients. Cultures were stimulated with either bacterial products such as lipo-oligosaccharide (LOS) or Staphylococcus aureus Cowan I (SAC) to activate monocytes or with monoclonal antibodies to CD3 and CD28 to induce polyclonal T-cell activation. We analysed the effect of methotrexate on cytokine production in these systems.Results. We showed that methotrexate inhibits production of cytokines induced by T-cell activation. Among the cytokines inhibited were interleukin 4 (IL-4), IL-13, IFNgamma, tumour necrosis factor-alpha (TNFalpha) and granulocyte-macrophage colony-stimulating factor. Inhibition was seen at concentrations easily achieved in plasma of RA patients taking the drug. IL-8 production was hardly influenced by methotrexate. Furthermore, inhibition was dependent on the stimulus; IL-6, IL-8, IL-1beta and TNFalpha production induced by LOS or SAC was only slightly decreased by methotrexate. The addition of folinic acid or thymidine and hypoxanthine reversed the inhibitory effects of methotrexate on cytokine production. Concentrations of methotrexate required for inhibition varied between donors. Oral intake of 10 mg methotrexate by RA patients led to marked inhibition of cytokine production in blood drawn after 2 h.Conclusions. Methotrexate turns out to be an efficient inhibitor of cytokine production induced by T-cell activation in freshly drawn blood. This is due to inhibition of the de novo synthesis of purines and pyrimidines. Cytokines produced by monocytes are hardly affected by methotrexate.