系统性红斑狼疮生物标志物的筛选及致病机制的研究

发布时间：2018-05-02 06:27

本文选题：系统性红斑狼疮 + 自身免疫抗体　；参考：《北京协和医学院》2017年博士论文

【摘要】：系统性红斑狼疮(Systemic lupus erythematosus,SLE)是一种经典的炎症性自身免疫性疾病,可导致全身多个脏器、系统受累,临床表现多样化,个体差异性大。目前,人们对SLE的发病机制认识并不是十分清楚,一般认为自身免疫抗体的产生后,与相应自身免抗原形成免疫复合物,沉积在皮肤、关节、肾脏等靶器官,激活补体,引起急慢性炎症反应及组织坏死所致。临床上,SLE主要依靠临床表现结合实验室检查来确诊,另外,也常常通过检测包括炎症因子和自身抗体等血清生物标志物来评估SLE的疾病活动性及治疗效果,但均不能理想地反映全部患者的病情进展及临床分类/分型情况。因此,探索和研究新的血清标记物及其理论基础,对于更好的评估SLE患者病情进展、指导患者的临床治疗及评估预后具有重要的意义。本研究首先通过Luminex高通量蛋白检测技术,对SLE患者血清中20种自身免疫抗体进行了检测,筛选出了SLE患者血清中特异性高表达的自身抗体、最佳联合诊断SLE的自身免疫抗体组合、与SLE疾病活动性相关的自身免疫抗体及与ANA、抗dsDNA抗体表达水平具有显著相关的自身免疫抗体。用层次聚类分析方法分析了这20种自身免疫抗体在SLE患者血清中的表达谱,发现同一种类的自身免疫抗体在SLE患者中的表达类型相似,常常联合出现。随后,通过血浆蛋白超滤浓缩冻干、Native-PAGE及免疫印迹等实验检测了SLE患者血液中自身免疫抗原抗体复合体的存在情况,证据支持在SLE患者血液中存在有大量的各种自身抗原及其抗体形成的抗原抗体复合物,由此,我们推测不同类型的SLE患者外周血中形成了不同的大分子免疫复合物,随血液循环进入到不同靶器官沉积致病。其中,抗U1小核糖核蛋白(1-small nuclear ribonucleoprotein,UlsnRNP)抗体与SLE肾脏受累具有显著相关性,我们用免疫荧光实验证实了免疫复合物沉积与其SLE患者在血清中表达水平、肾脏病理损害及肾脏功能损害一致,提示抗UlsnRNP抗体可作为SLE肾脏损伤的生物指标。另外,我们还发现在Ⅰ型IFN及Ⅰ型IFN诱导基因在SLE的发病中起重要作用,Ⅰ型IFN可作为评价SLE疾病活动性的生物学指标,IFI27、ISG15、SIGLEC1可作为LN的血清学指标。总之,本研究首次同步系统地分析了多种不同自身免疫抗体在SLE患者中的表达水平,筛选出了可以联合诊断SLE的血清学指标,并初步揭示了特异性自身免疫抗体以大分子抗原抗体免疫复合物存在于SLE患者的血液和组织,与特定靶器官病理损伤的关系。其中,鉴于抗UlsnRNP抗体与其抗原形成的免疫复合物沉积与狼疮性肾炎(Lupus nephritis,LN)相关性,我们提出了抗UlsnRNP抗体可作为临床SLE用来衡量LN肾脏损伤的血清学生物标志物,用于SLE的诊断及治疗效果评价。另外,我们还发现Ⅰ型IFN及其诱导的ISG在SLE发病中起着重要作用。上述研究的完成,进一步丰富可用于SLE诊断及治疗效果评价的生物标志物种类,也为我们认识和理解SLE发病机制提供了线索。
[Abstract]:Systemic lupus erythematosus (Systemic lupus erythematosus, SLE) is a classic inflammatory autoimmune disease, which can cause multiple organs in the whole body. The system is involved, the clinical manifestations are diverse and the individual difference is large. At present, people are not ten distinct in understanding the pathogenesis of SLE. Immune complexes are formed by self antigen free antigen, which are deposited in target organs such as skin, joints and kidneys, activating complement, causing acute and chronic inflammatory reactions and tissue necrosis. Clinically, SLE is mainly confirmed by clinical manifestations combined with laboratory tests. In addition, serum biomarkers, including inflammatory factors and autoantibodies, are often detected. To evaluate the disease activity and therapeutic effect of SLE, it is not ideal to reflect the progress of the patients and the clinical classification / classification. Therefore, it is important to explore and study the new serum markers and their theoretical basis for the better evaluation of the progression of SLE patients, the clinical treatment and the prognosis of the patients. In this study, 20 kinds of autoantibodies in serum of SLE patients were detected by Luminex high throughput protein detection technology, and the specific high expression of autoantibodies in serum of SLE patients was screened. The best combination of the autoimmune antibody combination of the diagnostic SLE, the autoimmune antibody related to the activity of SLE and the anti dsDNA resistance to the ANA, and the anti dsDNA resistance were found. A hierarchical cluster analysis method was used to analyze the expression profiles of these 20 autoimmune antibodies in the serum of SLE patients. It was found that the same type of autoimmune antibody in SLE patients was similar and often appeared together. Then, the plasma protein ultrafiltration was used to concentrate the freeze-dried, Native-PA GE and immunoblotting were used to detect the presence of autoantigen antibody complexes in the blood of SLE patients. The evidence supports the presence of a large number of various antigens and antibodies in the blood of SLE patients. Therefore, we speculate that different types of SLE patients have different macromolecules in the peripheral blood. The Phytophthora complex, with the circulation of blood circulation into different target organs, is deposited and pathogenic. Among them, the anti U1 1-small nuclear ribonucleoprotein (UlsnRNP) antibody and the SLE renal involvement have significant correlation. We confirmed the expression level of the immune complex in the serum of the SLE patients and the renal pathological damage by the immunofluorescence test. It is consistent with renal function damage, suggesting that anti UlsnRNP antibody can be used as a biomarker for SLE renal injury. In addition, we also found that type I IFN and type I IFN induced genes play an important role in the pathogenesis of SLE. Type I can be used as a biological indicator for evaluating the activity of SLE disease. IFI27, ISG15, SIGLEC1 can be used as a LN serological index. Anyway, In this study, the expression level of various autoimmune antibodies in SLE patients was analyzed synchronously for the first time. The serological indexes that could be combined with the diagnosis of SLE were screened, and the specific autoimmune antibodies were found in the blood and tissue of the SLE patients with the large molecular antigen antibody immune complex and the specific target organ pathology. In addition, in view of the correlation between the immune complex deposition of anti UlsnRNP antibody and its antigen formation and the Lupus nephritis (LN), we suggest that anti UlsnRNP antibody can be used as a serological biomarker for the clinical SLE used to measure the renal injury of LN, used in the diagnosis and evaluation of the therapeutic effect of SLE. In addition, we also send it. The present type I IFN and its induced ISG play an important role in the pathogenesis of SLE. The completion of these studies further enriches the species of biomarkers which can be used to evaluate the diagnostic and therapeutic effects of SLE, and also provides a clue to our understanding and understanding of the pathogenesis of SLE.

【学位授予单位】：北京协和医学院
【学位级别】：博士
【学位授予年份】：2017
【分类号】：R593.241

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