慢性乙型肝炎免疫耐受和免疫清除期淋巴细胞基因表达谱的差异

发布时间：2018-03-30 10:16

本文选题：慢性乙型肝炎　切入点：免疫耐受　出处：《天津医科大学》2006年硕士论文

【摘要】： 目的乙型肝炎病毒(HBV)是导致肝脏疾病的主要致病因子,许多的慢性乙型肝炎患者最终可发展成为肝硬化和原发性肝癌。迄今为止,乙型肝炎的发病机理和HBV引发免疫耐受的机制尚未完全阐明。本课题研究慢性乙型肝炎患者免疫耐受期和免疫清除期体内淋巴细胞基因表达谱的差异,探讨差异表达基因与HBV感染引起的免疫耐受及免疫清除的关系。方法选择慢性乙型肝炎(HBskg、HBekg、HBcAb阳性)病例,根据丙氨酸氨基转移酶(ALT)高低划分免疫耐受组、免疫清除组;以健康查体者为正常对照组。采集新鲜血液,EDTA抗凝,淋巴细胞分离液分离淋巴细胞,一步法提取总RNA,用逆转录法标记cDNA探针,荧光染料Cy3标记正常对照组,Cy5标记免疫耐受组和免疫清除组。采用高通量基因表达谱芯片(7,267个基因)进行芯片杂交,使用ScanArray Express共聚焦扫描仪扫描基因芯片,PerkinElmer LAS软件进行数据分析。结果比较免疫耐受组与正常对照组发现差异表达基因419个,表达上调206个,下调213个;免疫清除组与免疫耐受组差异表达基因共213个,上调111个,下调102个;两组共有但差异表达方向相反的基因47个,在免疫耐受期表达下调但免疫清除期上调6个,在免疫耐受上调免疫清除期下调41个。结论使用表达谱基因芯片检测慢性乙型肝炎患者免疫耐受、免疫清除期淋巴细胞的基因表达,结合HBV感染的自然史,初步建立了HBV免疫耐受及免疫清除发生时淋巴细胞的差异表达基因谱。通过比对免疫耐受和免疫清除发生后基因表达谱的改变,筛选出47个发生反向差异表达的基因。这些基因的反向差异表达可能在HBV的免疫耐受和免疫清除中的发挥了重要的作用。其一方面揭示了HBV逃避宿主免疫系统清除时对淋巴细胞产生的主动性调节机制,另一方面反映了免疫清除发生时机体淋巴细胞应答调机制。
[Abstract]:Purpose. Hepatitis B virus (HBV) is a major cause of liver disease, and many patients with chronic hepatitis B can eventually develop cirrhosis and primary liver cancer. The pathogenesis of hepatitis B and the mechanism of immune tolerance induced by HBV have not been fully elucidated. To investigate the relationship between differentially expressed genes and immune tolerance and clearance induced by HBV infection. Method. According to the level of alanine aminotransferase (alt), the immune tolerance group was divided into immune tolerance group and immune clearance group, and the healthy examiner was used as the normal control group, and the fresh blood EDTA anticoagulant was collected. Lymphocytes were isolated from lymphocytes, total RNAs were extracted by one step method, and cDNA probe was labeled with reverse transcription method. Fluorescent dye Cy3 was used to label Cy5 in normal control group and immune tolerance group and clearance group. High throughput gene expression microarray (7267 genes) was used for microarray hybridization. ScanArray Express confocal scanner was used to scan gene chip Perkin Elmer LAS for data analysis. Results. There were 419 differentially expressed genes (206 up-regulated and 213 down-regulated) in the immune tolerance group and the normal control group, 213 differentially expressed genes were up-regulated and 102 down-regulated in the immune clearance group and the immune tolerance group. There were 47 genes in the two groups, which were down-regulated in immune tolerance phase but up-regulated in immune clearance stage, and 41 genes were down-regulated in immune tolerance up-regulated immune clearance phase. Conclusion. Expression profile gene microarray was used to detect immune tolerance and lymphocyte gene expression in patients with chronic hepatitis B, and combined with the natural history of HBV infection. The differentially expressed gene profiles of lymphocytes during HBV tolerance and immune clearance were preliminarily established. By comparing the changes of gene expression profiles after immune tolerance and immune clearance, 47 reverse differentially expressed genes were screened. The reverse differential expression of these genes may play an important role in the immune tolerance and clearance of HBV. On the one hand, it reveals that HBV evades the host immune system. The mechanism of active regulation of lymphocyte production, On the other hand, it reflects the mechanism of lymphocyte response regulation during immune clearance.
【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R512.62;R392.1

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