Hückel Marion, Schurigt Uta, Wagner Andreas H, Stöckigt Renate, Petrow Peter K, Thoss Klaus, Gajda Mieczyslaw, Henzgen Steffen, Hecker Markus, Bräuer Rolf
Institute of Pathology, Friedrich Schiller University, Jena, Germany.
Arthritis Res Ther. 2006;8(1):R17. doi: 10.1186/ar1869.
The transcription factor STAT-1 (signal transducer and activator of transcription-1) plays a pivotal role in the expression of inflammatory gene products involved in the pathogenesis of arthritis such as various cytokines and the CD40/CD40 ligand (CD40/CD40L) receptor-ligand dyad. The therapeutic efficacy of a synthetic decoy oligodeoxynucleotide (ODN) binding and neutralizing STAT-1 was tested in murine antigen-induced arthritis (AIA) as a model for human rheumatoid arthritis (RA). The STAT-1 decoy ODN was injected intra-articularly in methylated bovine serum albumin (mBSA)-immunized mice 4 h before arthritis induction. Arthritis was evaluated by joint swelling measurement and histological evaluation and compared to treatment with mutant control ODN. Serum levels of pro-inflammatory cytokines, mBSA-specific antibodies and auto-antibodies against matrix constituents were assessed by enzyme-linked immunosorbent assay (ELISA). The transcription factor neutralizing efficacy of the STAT-1 decoy ODN was verified in vitro in cultured synoviocytes and macrophages. Single administration of STAT-1 decoy ODN dose-dependently suppressed joint swelling and histological signs of acute and chronic arthritis. Delayed-type hypersensitivity (DTH) reaction, serum levels of interleukin-6 (IL-6) and anti-proteoglycan IgG titres were significantly reduced in STAT-1 decoy ODN-treated mice, whereas mBSA, collagen type I and type II specific immunoglobulins were not significantly affected. Intra-articular administration of an anti-CD40L (anti-CD154) antibody was similarly effective. Electrophoretic mobility shift analysis (EMSA) of nuclear extracts from synoviocytes incubated with the STAT-1 decoy ODN in vitro revealed an inhibitory effect on STAT-1. Furthermore, the STAT-1 decoy ODN inhibited the expression of CD40 mRNA in stimulated macrophages. The beneficial effects of the STAT-1 decoy ODN in experimental arthritis presumably mediated in part by affecting CD40 signalling in macrophages may provide the basis for a novel treatment of human RA.
转录因子STAT-1（信号转导及转录激活因子1）在参与关节炎发病机制的炎性基因产物表达中起关键作用，这些炎性基因产物包括多种细胞因子以及CD40/CD40配体（CD40/CD40L）受体 - 配体二元体。作为人类类风湿关节炎（RA）的模型，在小鼠抗原诱导性关节炎（AIA）中测试了一种结合并中和STAT-1的合成诱饵寡脱氧核苷酸（ODN）的治疗效果。在诱导关节炎前4小时，将STAT-1诱饵ODN关节内注射到经甲基化牛血清白蛋白（mBSA）免疫的小鼠体内。通过测量关节肿胀和组织学评估来评价关节炎，并与用突变对照ODN治疗的情况进行比较。通过酶联免疫吸附测定（ELISA）评估促炎细胞因子、mBSA特异性抗体和针对基质成分的自身抗体的血清水平。在体外培养的滑膜细胞和巨噬细胞中验证了STAT-1诱饵ODN的转录因子中和效果。单次给予STAT-1诱饵ODN剂量依赖性地抑制关节肿胀以及急性和慢性关节炎的组织学体征。在接受STAT-1诱饵ODN治疗的小鼠中，迟发型超敏反应（DTH）、白细胞介素-6（IL-6）血清水平和抗蛋白聚糖IgG滴度显著降低，而mBSA、I型和II型胶原特异性免疫球蛋白未受到显著影响。关节内给予抗CD40L（抗CD154）抗体同样有效。体外将STAT-1诱饵ODN与滑膜细胞核提取物一起孵育后的电泳迁移率变动分析（EMSA）显示对STAT-1有抑制作用。此外，STAT-1诱饵ODN抑制刺激的巨噬细胞中CD40 mRNA的表达。STAT-1诱饵ODN在实验性关节炎中的有益作用可能部分是通过影响巨噬细胞中的CD40信号传导介导的，这可能为人类RA的新治疗方法提供基础。
