【作者】 赵帅；

【导师】 方艳秋； 付嘉；

【作者基本信息】 吉林大学 ， 免疫学， 2010， 硕士

【摘要】 抗磷脂抗体综合征(Antiphospholipid antibody syndrome, APS)是一种以习惯性流产、动静脉血栓形成为主要特征的一类累及多器官、系统的自身免疫病,患者血中存在抗磷脂抗体(Antiphospholipid antibody, APA)。现已知β2糖蛋白1(β2-glycoprotein-1,β2-GP1)是高度糖基化的单链血浆蛋白,是APA (anti-β2GP I)主要的靶抗原。大量证据显示抗β2糖蛋白1抗体(anti-beta2-glycoprotein-1 antibody, anti-β2GP I)、抗心磷脂抗体(anticardiolipin antibodies, ACA)以致炎因子生成、内皮细胞和血小板活化、凝血因子和补体激活等机制参与APS的发病。CD4+CD25+调节性T细胞(CD4+CD25+regulatory T cell, Treg)是调节性T细胞中最为重要的一群,因其具有免疫调节和抑制作用,近年来受到研究者的密切关注。研究表明调节性T细胞对于维持机体免疫自稳,防止自身免疫病具有极其重要的作用,资料显示CD4+CD25+Treg的数量减少或功能异常可与多种自身免疫病相关。但对于在APS及APS实验动物模型中CD4+CD25+Treg功能的变化国内外却鲜见报道。本实验将以重组人p2糖蛋白1(recombinant humanβ2-glycoprotein-1, rhβ2GP I)主动免疫BALB／c小鼠建立实验性APS模型,采用流式细胞术检测小鼠PBMC中CD4+CD25+Treg细胞数目,RT-PCR检测转录因子Foxp3 mRNA的表达,以观察CD4+CD25+Treg细胞数目变化及活性变化是否参与APS的发病机制。结果发现,在应用rhβ2GPⅠ免疫小鼠12w后,CD4+CD25+Treg细胞亚群的比例及Foxp3 mRNA表达开始明显降低,这与rhβ2GPⅠ免疫小鼠开始出现APS疾病的临床征象时间一致。提示CD4+CD25+Treg细胞在机体内起重要的免疫调节作用,正常情况下它与效应性T细胞互相拮抗抑制,随着CD4+CD25+Treg细胞数量及活性下降后抑制作用减弱,表现出效应性T细胞发育旺盛从而表现出APS的临床征象。CD4+CD25+Treg细胞数量的多少及活性的高低是发生APS的重要发病因素。我们期待CD4+CD25+Treg细胞数量的多少及活性的高低能否成为日后诊断APS的辅助诊断依据,通过监测CD4+CD25+Treg细胞数量的多少及活性的高低能否成为预防APS的检验手段,以及患有APS疾病的人们出现CD4+CD25+Treg细胞数量的减少及活性的降低,输注CD4+CD25+Treg细胞能否成为有效的治疗手段。随着对APS发病机制的深入研究,人类将找到有效的检测及治疗方法。更多还原

【Abstract】 Objective:To observe the quantitative and functional changes of CD4+CD25+regulatory cells in mice during the development of experimental anti-phospholipid antibody syndrome (EAPS). Methods:To establish the model of EAPS, we immunized BALB/c mice with rhβ2-GP1. Twelve weeks after immunization, aCL、anti-β2GPI titers in the sera, the percentage of the fetal resorptions, activated partial thromboplastin time (aPTT) and platelet counts were assayed. Frequency of CD4+CD25+Treg in peripheral blood mononuclear cells from EAPS mice were determined by flow cytometry, mRNA expression of foxp3 were semi-quantified by RT-PCR. Results:Model mice showed an elevated titers of anti-β2GPI and aCL Abs in the sera (p<0.05), higher resorption percentage (p<0.05), prolonged aPTT (p<0.05), and lower platelet counts (p<0.05). The expressing levels of foxp3 mRNA in the model group was higher than those of control group (p<0.05) in the fourth week after immunization, but the expressing level of Foxp3 mRNA began to decrease in week 8 and were lower than those of control group from week twelve (p<0.05). The frequency of CD4+CD25+Treg cells in model group was similar with that of control group within the eighth week after immunization (p>0.05), but after week twelve, the frequency of CD4+CD25+Treg cells in model group began to get lower than contol group (p<0.05).Conclusion:The decreased number and function of CD4+CD25+Treg cells may contribute to the pathogenic mechanism in EAPS.更多还原