Lewis Katz School of Medicine at Temple University, Philadelphia, Pennsylvania.
University of North Carolina at Chapel Hill.
Arthritis Rheumatol. 2016 Nov;68(11):2697-2707. doi: 10.1002/art.39733.
Increased levels of type I interferon (IFN) and type I IFN-regulated genes are found in patients with systemic lupus erythematosus (SLE) and may be central to its pathogenesis. Mitochondrial antiviral signaling protein (MAVS) is a key regulator of type I IFN that undergoes a dramatic prion-like aggregation and self propagates the activation signal from viral RNA to amplify downstream IFN production. We undertook this study to determine whether such MAVS aggregates might play a role in the sustained increased production of type I IFN in SLE.
Peripheral blood mononuclear cells were isolated and mitochondrial extracts were prepared. MAVS aggregation was detected by semidenatured agarose gel electrophoresis and confirmed by immunofluorescence staining. MAVS-associated signaling proteins were analyzed by Western blotting. MAVS aggregation-associated gene expression signature was analyzed by microarray.
In blood cells from 22 of 67 SLE patients, essentially all MAVS was in a high molecular weight aggregated form. None of 6 rheumatoid arthritis patients and only 3 of 33 healthy controls had abnormal MAVS. Compared to MAVS aggregate-negative patients, MAVS aggregate-positive SLE patients had significantly higher serum levels of IFNβ and significantly increased levels of autoantibodies against Sm and U1 RNP. Gene array data revealed a characteristic gene expression pattern in these patients, with altered expression of genes involved in IFN signaling and membrane trafficking.
Persistent MAVS aggregates may lead to increased type I IFN production and result in unmitigated signals leading to autoimmunity.
红斑狼疮(SLE)患者体内存在Ⅰ型干扰素(IFN)及其Ⅰ型 IFN 调控基因水平升高的现象,这可能是其发病机制的核心。线粒体抗病毒信号蛋白(MAVS)是Ⅰ型 IFN 的关键调节因子,其经历剧烈的类朊病毒聚集,并自我传播病毒 RNA 激活信号,放大下游 IFN 的产生。我们进行此项研究旨在确定这种 MAVS 聚集是否可能在 SLE 患者持续产生大量Ⅰ型 IFN 中发挥作用。
分离外周血单核细胞并制备线粒体提取物。通过非变性琼脂糖凝胶电泳检测 MAVS 聚集,并通过免疫荧光染色进行验证。通过 Western 印迹分析 MAVS 相关信号蛋白。通过微阵列分析 MAVS 聚集相关基因表达谱。
在 67 例 SLE 患者的 22 例血液细胞中,几乎所有的 MAVS 均处于高分子量聚集形式。6 例类风湿关节炎患者和 33 例健康对照者中均无异常 MAVS。与 MAVS 聚集阴性患者相比,MAVS 聚集阳性 SLE 患者的 IFNβ 血清水平显著升高,且针对 Sm 和 U1 RNP 的自身抗体水平显著升高。基因芯片数据显示这些患者存在特征性的基因表达模式,IFN 信号和膜转运相关基因的表达发生改变。
持续的 MAVS 聚集可能导致Ⅰ型 IFN 产生增加,并导致自身免疫的信号无法被抑制。