乙肝病毒相关肝病患者肝移植后外周血单个核细胞及肝组织中乙型肝炎病毒cccDNA的检测  

Detection of hepatitis B virus cccDNA in peripheral blood monuclear cells and liver tissues in hepatitis B virus-related disease patients after liver transplantation

作　　者：张荣[1] 吴忠均[1] 曹辰[1] 陈映[1] 

机构地区：[1]重庆医科大学第一附属医院,重庆市400016

出　　处：《中国组织工程研究与临床康复》2011年第5期783-787,共5页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：国家自然科学基金(30772055)项目;重庆市卫生局科研项目(06-2-017)课题名称:乙肝相关性肝病肝移植术后乙肝复发的机制及干预研究~~

摘　　要：背景:肝移植后受者体内绝大部分病毒负荷被清除,植入新肝后其复发性肝炎病原体从肝外进入肝内的途径及其复制规律目前尚无定论。目的:检测肝移植前后外周血单个核细胞和肝组织中乙型肝炎病毒cccDNA及血清中乙型肝炎病毒DNA的表达。方法:采用淋巴细胞分离液从乙肝病毒相关终末期肝病37例患者外周血中分离出单个核细胞,采用荧光定量PCR检测肝移植前后及移植后乙肝复发3个时期外周血单个核细胞和肝组织中cccDNA及血清乙型肝炎病毒DNA表达。结果与结论:肝移植前,单个核细胞cccDNA阳性12例,肝组织cccDNA阳性6例,检出率分别为32%和16%,单个核细胞、肝组织中cccDNA拷贝范围分别为(3.028～6.508)×104,(4.158～6.234)×104拷贝/mL。肝移植后,单个核细胞cccDNA阳性1例,无血清乙型肝炎病毒DNA检测阳性病例。6例肝移植后乙肝复发病例中外周血单个核细胞cccDNA阳性4例,肝组织活检cccDNA阳性1例,6例血清乙型肝炎病毒DNA均为阳性。提示乙肝病毒相关终末期肝病患者肝移植后乙肝复发途径可能是残留乙肝病毒在外周单个核细胞中以cccDNA为模板复制,然后再迁移到肝脏。BACKGROUND：Life-long prophylaxis against hepatitis B virus （HBV） recurrence is recommended in patients transplanted for HBV-related disease. But exact mechanism of HBV recurrence and its copy law is unknown. OBJECTIVE：By detection of HBV cccDNA and HBV DNA in peripheral blood mononuclear cells （PBMCs） and liver tissues and serum from HBV-related end-stage liver disease patients before and after liver transplantation. METHODS：PBMCs were isolated from HBV-related end-stage liver disease patients by lymphocyte separation medium. HBVcccDNA in PBMCs and liver tissue and the serum HBVDNA at three periods of before,after liver transplantation and postoperative recurrence were detected by fluorescent quantitative PCR. RESULTS AND CONCLUSION：Before liver transplantation,cccDNA in PBMCs and liver tissues were positive in 12 cases and 6 cases,the detection rates were 32.43% and 16.22%,cccDNA copies in PBMCs ranged from 3.028 to 6.508×104 copies/mL,as in liver tissues the range was from 4.158 to 6.234×104 copies/mL. After liver transplantation,cccDNA in PBMCs was positive in one case. In 6 cases of hepatitis B recurrence after liver transplantation,cccDNA in PBMCs was positive in 4 cases,as positive in one case of liver biopsy,and all 6 cases positive for serum HBVDNA. Possible way of hepatitis B recurrence of HBV-related end-stage liver disease patients after liver transplantation may be that residual HBV in peripheral mononuclear cells replicate according to cccDNA as the template,and then migrate to the liver.

关 键 词：乙型肝炎病毒 乙型肝炎病毒CCCDNA 单个核细胞 肝移植 乙肝复发 

分 类 号：R617[医药卫生—外科学]