Center for Molecular and Cellular Intervention, Department of Pediatric Immunology, Wilhelmina Children's Hospital, University Medical Centre Utrecht, Utrecht, The Netherlands
Center for Molecular and Cellular Intervention, Department of Pediatric Immunology, Wilhelmina Children's Hospital, University Medical Centre Utrecht, Utrecht, The Netherlands.
Rheumatology (Oxford). 2015 Sep;54(9):1724-34. doi: 10.1093/rheumatology/kev101. Epub 2015 Apr 14.
The balance between Treg and effector T cells (Teff) is crucial for immune regulation in JIA. How MTX, the cornerstone treatment in JIA, influences this balance in vivo is poorly elucidated. The aim of this study was to investigate quantitative and qualitative effects of MTX on Treg and Teff in JIA patients during MTX treatment.
Peripheral blood samples were obtained from JIA patients at the start of MTX and 3 and 6 months thereafter. Treg numbers and phenotypes were determined by flow cytometry and suppressive function in allogeneic suppression assays. Teff proliferation upon stimulation with anti-CD3, activation status and intracellular cytokine production were determined by flow cytometry. Effector cell responsiveness to suppression was investigated in autologous suppression assays. Effector cell cytokines in supernatants of proliferation and suppression assays and in plasma were measured by cytokine multiplex assay.
MTX treatment in JIA did not affect Treg phenotype and function. Instead, MTX treatment enhanced, rather than diminished, CD4(+) and CD8(+) T cell proliferation of JIA patients after 6 months of therapy, independent of clinical response. Effector cells during MTX treatment were equally responsive to Treg-mediated suppression. MTX treatment did not attenuate Teff activation status and their capacity to produce IL-13, IL-17, TNF-α and IFN-γ. Similarly to Teff proliferation, plasma IFN-γ concentrations after 6 months were increased.
This study provides the novel insight that MTX treatment in JIA does not attenuate Teff function but, conversely, enhances T cell proliferation and IFN-γ plasma concentrations in JIA patients.
调节性 T 细胞（Treg）与效应 T 细胞（Teff）之间的平衡对于 JIA 的免疫调节至关重要。甲氨蝶呤（MTX）作为 JIA 的基石治疗药物，其在体内如何影响这种平衡尚未得到充分阐明。本研究旨在探讨 MTX 治疗期间 MTX 对 JIA 患者 Treg 和 Teff 的定量和定性影响。
在开始 MTX 治疗时以及之后的 3 个月和 6 个月，从 JIA 患者中获得外周血样本。通过流式细胞术测定 Treg 数量和表型，并在同种异体抑制试验中测定其抑制功能。通过流式细胞术测定 Teff 在抗 CD3 刺激下的增殖、激活状态和细胞内细胞因子产生。通过自体抑制试验研究效应细胞对抑制的反应性。通过细胞因子多重分析测定增殖和抑制试验上清液及血浆中效应细胞细胞因子。
MTX 治疗 JIA 不会影响 Treg 表型和功能。相反，6 个月的治疗后，MTX 治疗增强了而不是减弱了 JIA 患者的 CD4(+)和 CD8(+)T 细胞增殖，与临床反应无关。MTX 治疗期间的效应细胞对 Treg 介导的抑制同样敏感。MTX 治疗并未减弱 Teff 的激活状态及其产生 IL-13、IL-17、TNF-α和 IFN-γ的能力。与 Teff 增殖一样，6 个月后 IFN-γ 血浆浓度也增加。
本研究提供了新的见解，即 MTX 治疗 JIA 不会减弱 Teff 功能，而是相反，增强了 JIA 患者的 T 细胞增殖和 IFN-γ 血浆浓度。
