Department of Neurology, Xiangya Hospital, Central South University, 410008, Changsha, Hunan, People's Republic of China.
Inflamm Res. 2010 Mar;59(3):197-205. doi: 10.1007/s00011-009-0087-6. Epub 2009 Sep 19.
To observe the therapeutic effect of RelB-silenced dendritic cells (DCs) in experimental autoimmune myasthenia gravis (EAMG), and further to investigate the mechanism of this immune system therapeutic.
(1) RelB-silenced DCs and control DCs were prepared and the supernatants were collected for IL-12p70, IL-6, and IL-23 measurement by ELISA. (2) RelB-silenced DCs and control DCs were co-cultured with AChR-specific T cells, and the supernatant was collected to observe the IL-17, IFN-gamma, IL-4 production. (3) EAMG mice with clinical scores of 1 to 2 were collected and enrolled randomly into the RelB-silenced DC group or the control DC group. RelB-silenced DCs or an equal amount of control DCs were injected intravenously on days 0, 7, and 14 after enrollment. Clinical scores were evaluated every other day. Twenty days after allotment, serum from individual mice was collected to detect serum concentrations of anti-mouse AChR IgG, IgG1, IgG2b, and IgG2c. The splenocytes were isolated for analysis of lymphocyte proliferative responses, cytokine (IL-17, IFN-gamma, IL-4) production, and protein levels of RORgammat, T-bet, GATA-3, and FoxP3 (the special transcription factors of Th17, Th1, Th2, and Treg, respectively).
(1) RelB-silenced DCs produced significantly reduced amounts of IL-12p70, IL-6, and IL-23, as compared with control DCs. (2) RelB-silenced DCs spurred on the CD4(+) T cells from Th1/Th17 to the Th2 cell subset in the co-culture system. (3) Treatment with RelB-silenced DCs effectively reduced myasthenic symptoms and levels of serum anti-acetylcholine receptor autoantibody in mice with ongoing EAMG. Th17-related markers (RORgammat, IL-17) and Th1-related markers (T-bet, IFN-gamma) were downregulated, whereas Th2 markers (IL-4 and GATA3) and Treg marker (FoxP3) were upregulated.
RelB-silenced DCs were able to create a particular cytokine environment that was absent of inflammatory cytokines. RelB-silenced DCs provide a practical means to normalize the differentiation of the four T-cell subsets (Th17, Th1, Th2, and Treg) in vivo, and thus possess therapeutic potential in Th1/Th17-dominant autoimmune disorders such as myasthenia gravis.
观察 RelB 沉默树突状细胞（DC）在实验性自身免疫性重症肌无力（EAMG）中的治疗效果，并进一步探讨这种免疫系统治疗的机制。
（1）制备 RelB 沉默 DC 和对照 DC，并通过 ELISA 检测上清液中 IL-12p70、IL-6 和 IL-23 的含量。（2）将 RelB 沉默 DC 和对照 DC 与 AChR 特异性 T 细胞共培养，收集上清液观察 IL-17、IFN-γ、IL-4 的产生。（3）收集临床评分 1 至 2 的 EAMG 小鼠，随机分为 RelB 沉默 DC 组或对照 DC 组。在入组后第 0、7 和 14 天静脉注射 RelB 沉默 DC 或等量对照 DC。每隔一天评估临床评分。分配后 20 天，从个体小鼠中采集血清，检测血清中抗小鼠乙酰胆碱受体 IgG、IgG1、IgG2b 和 IgG2c 的浓度。分离脾细胞分析淋巴细胞增殖反应、细胞因子（IL-17、IFN-γ、IL-4）产生以及转录因子（RORγt、T-bet、GATA-3、FoxP3，分别为 Th17、Th1、Th2 和 Treg 的特殊转录因子）的蛋白水平。
（1）与对照 DC 相比，RelB 沉默 DC 产生的 IL-12p70、IL-6 和 IL-23 明显减少。（2）RelB 沉默 DC 在共培养系统中刺激 CD4+T 细胞从 Th1/Th17 向 Th2 细胞亚群分化。（3）用 RelB 沉默 DC 治疗可有效减轻正在发生的 EAMG 小鼠的肌无力症状和血清抗乙酰胆碱受体自身抗体水平。Th17 相关标志物（RORγt、IL-17）和 Th1 相关标志物（T-bet、IFN-γ）下调，而 Th2 标志物（IL-4 和 GATA3）和 Treg 标志物（FoxP3）上调。
RelB 沉默 DC 能够产生缺乏炎症细胞因子的特定细胞因子环境。RelB 沉默 DC 为体内调节四群 T 细胞（Th17、Th1、Th2 和 Treg）的分化提供了一种实用手段，因此在 Th1/Th17 占优势的自身免疫性疾病如重症肌无力中具有治疗潜力。
