肝移植术后肝组织中HBV cccDNA的表达及临床意义

目的探讨肝移植术后患者肝组织内乙型肝炎病毒共价闭合环状DNA(HBV cccDNA)的表达及其临床意义。方法应用原位聚合酶链反应及免疫组织化学法检测2004年4月至2007年4月在北京地坛医院肝移植术后出现HBV再感染的25例患者及10例肝移植术后未出现HBV再感染者肝组织中HBV cccDNA及乙型肝炎表面抗原(HBsAg)、乙型肝炎核心抗原(HBcAg)、前-S1的表达。结果在35例标本中检测到cccDNA阳性的有22例(62.9%),阳性信号为紫蓝色呈块状或颗粒状,定位于细胞核。25例术后出现HBV再感染的患者其HBV cccDNA阳性率显著高于未出现HBV再感染者(84%对10%,P<0.01)。HBV cccDNA和HBcAg均为阳性的有20例,有2例患者肝组织中HBcAg阴性,但HBV cccDNA检测为阳性。肝组织中HBV cccDNA的表达阳性率与HBcAg具有高度一致性(Kappa=0.755,P<0.01)。结论肝移植患者肝组织中HBV cccDNA阳性表达可能与乙型肝炎再感染具有密切关系。

Clinical significance and expression of HBV cccDNA in liver specimens from patients after liver transplantation.

Objective To develop a sensitive and specific in situ assay for detecting HBV cccDNA in biopsy specimens,to detect intrahepatic in situ HBV cccDNA of patients with liver transplantation and to explore its clinical significance.Methods Intrahepatic in situ cccDNA and HBsAg,HBcAg,pre-S1 in liver biopsy specimens of 25 patients with HBV re-infection and 10 patients without HBV re-infection after liver transplantation were detected by in situ polymerase chain reaction(in situ PCR)and immunohistochemistry,respectively.Results In all 35 cases detected by in situ PCR,22 showed HBV cccDNA positive(62.9%).Positive signal presented as purple blue block or granules located in affected hepatocyte nuclei rather than cytoplasm or membrane.The positive rate of cccDNA in HBV re-infection group was greatly higher than that in control group(84% vs 10%,P<0.01).Detection of HBV cccDNA and HBcAg in liver biopsy specimens had shown high consistency(Kappa=0.755,P<0.01).Conclusion The expression of HBV cccDNA in liver biopsy specimens from the patients after liver transplantation may be closely associated with HBV re-infection.