乙型肝炎病毒急性感染小鼠模型的建立

采用高压水注射方法,通过尾静脉将具有复制能力的HBV质粒导入BABL/cJ小鼠体内,应用real-time PCR、ELISA、RIA、Southern Blot、Northern Blot,以及免疫组化等方法,检测小鼠病毒血症、血清和肝组织中HBV 抗原表达动态变化、肝组织中HBV转录和复制情况,以及小鼠免疫应答状况。结果HBV基因可以在小鼠体内表 达和复制,并诱导小鼠产生特异性免疫应答,其应答模式及HBV清除过程与人类的HBV急性感染类似。实验显 示高压注射具有复制能力的HBV质粒可以在小鼠体内建立HBV急性感染模型,这种模型可以用于HBV病毒 学、免疫学以及抗病毒药物筛选等方向的研究。

Development of a Mouse Model for Acute Hepatitis B Virus Infection

Using hydrodynamic injection. pUC18-HBV1. 3 containing a replication-competent, HBV1.3 supergenomic DNA, was injected into BALB/cJ mice through tail vain. HBV viremia was measured by real-time PCR. Serum concentrations of HBsAg were detected by sandwich ELISA HBeAg was detected and quantified by RIA using HBcAg standards to generate. Antibodies specific for HBsAg, and HBcAg were assayed by endpoint titration ELISA. HBV core protein expressed in the liver was visualized by immunohistochemical staining. Viral RNA and replicative DNA intermediates in liver were detected by Northern and Southern blot analyses, respectively. Hydrodynamic injection of pUC18-HBV1. 3 lead to viral gene expression and replication in vivo . The immune response to HBV. which was comparable with the characteristic response during acute HBV infection in man, was elicited in mice. A mouse model of acute hepatitis B virus infection can be developed by transfection of hepatocytes in vivo .