节点文献
慢性乙肝患者外周血单个核细胞FOXP3 mRNA表达的初步分析
Real-time RT-PCR for Detection of FOXP3 Expression in Peripheral Blood Mononuclear Cells of Patients with Hepatits B
【作者】 郭凯；
【导师】 黄长形；
【作者基本信息】 第四军医大学 ， 内科学， 2008， 硕士
【摘要】 乙型肝炎病毒(hepatitis B virus,HBV)慢性感染是非常严重的人类健康问题,对HBV感染慢性化发生机制的研究是寻找新的有效的治疗方法的基础。几十年来世界各国的科学家就此进行了大量的研究,发现特异性免疫功能低下是HBV感染慢性化的关键,但是未能阐明免疫功能低下发生的机制。目前认为维持机体免疫耐受的主要机制是中枢耐受和外周耐受,在外周耐受中调节性T细胞(Treg细胞)具有非常重要的作用,已成为当前免疫学研究的热点。国内外多项研究结果显示调节性T细胞与慢性HBV感染者特异性免疫反应低下有一定的关联。国外多个研究小组已证明FOXP3(称叉头状/翼状螺旋转录因子)在Treg细胞上特异性表达,且对Treg细胞的发育和功能是必需的。这极大地促进了在分子水平上对Treg细胞的认识,但当前对于FOXP3是如何调控Treg细胞的发育及其通过何种下游靶基因发生作用的机制仍知之甚少。阐明这些问题,对寻找以调节性T细胞为靶点的新的治疗自身免疫性疾病、肿瘤和包括慢性乙型病毒性肝炎在内的慢性感染性疾病具有重要意义。本实验研究目的是分析慢性乙肝患者外周血单个核细胞(PBMC)中FOXP3 mRNA的表达,比较乙肝患者与健康人PBMC中FOXP3 mRNA表达水平差异,初步阐明其在HBV慢性感染免疫机制中的作用,为临床观察慢性乙型肝炎患者免疫功能提供一个潜在指标,并为深入研究HBV慢性感染与FOXP3表达之间的关系奠定基础。方法:利用实时荧光定量RT-PCR的方法检测25例慢性乙肝患者以及11名健康人PBMC中FOXP3 mRNA的水平。结果与结论:乙肝患者PBMC中FOXP3 mRNA的表达水平高于健康对照组(8.41±0.13vs9.82±0.20),差异具有统计学意义(P﹤0.01),且与血清中的病毒含量呈正相关(r=0.70,P<0.01),而与ALT水平无关(r=0.06,P>0.05)。慢性乙肝患者FOXP3表达水平可能是HBV感染持续的一个因素。
【Abstract】 Chronic HBV infection is one of the major health concerns around the world. The understanding on the mechanism of chronic HBV infection is the foundation for developing a novel effective treatment. The research results indicate that the suppression of specific immune function is a key factor for HBV chronic infection. However, the machanism of this suppressive immune response still remains unclear. The maintainance of immunotolerance is considered to depend on central tolerance and peripherial tolerance, and the regulatory T cell (Treg) has great role in maintaining immune tolerance and stablizing immune response. Treg cell is related to the low specific immune-response in chronic HBV infection patients.FOXP3(the forkhead/winged-helix transcriptional regulators P3) is expressed specifically in regulatory T cells, and exerts great influence on the development and function of Treg cells. Currently, however, very little is known about the mechanism of FOXP3 regulates the development and function of Treg cells. The elucidation on the problems is very important for developing new treatments targetd on Treg for autoimmune diseases, tumors, and chornic infectious diseases including chornic hepatitis B. In this study, the objective is to examine the relationship between FOXP3 mRNA level in peripheral blood mononuclear cell(PBMC) and hepatitis B infection.Methods: Real-time fluorescence relative quantitative RT-PCR was used to quantify FOXP3 mRNA levels in peripheral blood mononuclear cells from 11 HBV-uninfected subjects and 25 patients with chronic hepatitis B disease.Results and Conclusions: FOXP3 mRNA levels in hepatitis B patients were higher than that in HBV-uninfected subjects (8.41±0.13vs9.82±0.20,P<0.01) and were correlated with HBV viral loads directly(r=0.70,P<0.01),but were not correlated with ALT(r=0.06,P>0.05). High expression of FOXP3 might be a important factor for persistence of HBV infection.