重组8型腺相关病毒介导HBV急性感染树鼩模型建立

目的利用重组8型腺相关病毒介导1.3拷贝HBV基因组（1.3HBV,ayw亚型）在树鼩肝脏表达,建立HBV急性感染树鼩模型。方法通过大腿内侧静脉注射将携带有1.3 HBV的重组8型腺相关病毒（recombi-nant adeno-associated virus 8,rAAV8-1.3HBV）导入树鼩肝脏,通过ELISA检测树鼩血清中HBsAg、HBeAg、HBsAb、HBeAb、HBcAb,荧光定量PCR检测树鼩肝脏和血清中HBV DNA,全自动生化分析仪检测血清中ALT水平,并观察感染后肝脏的病变情况。结果 HBV感染主要血清标志物1～2周内均检测阳性;30 d后肝组织仍可检测到病毒抗原阳性细胞;55 d时肝组织HBV DNA拷贝数仍可达到104～105;树鼩血清中HBV DNA拷贝数持续一个月高于正常组;肝组织炎细胞略增多,血清ALT水平持续升高。结论 rAAV8所携带的HBV基因组高效专一导入树鼩肝细胞并复制表达,成功建立HBV急性感染树鼩模型,为进一步探索rAAV8树鼩慢性感染模型打下一定的基础。

Establishment of a tree shrew infection by transduction with virus 8 carrying 1.3 model of acute hepatitis B virus a recombinant adeno-associated copies of HBV genome

Objective To establish a tree shrew model of acute hepatitis B virus infection by injection of a recombinant adeno-assoeiated virus 8 vector carrying 1.3 copies of HBV genome （ayw subtype） （rAAV8-1.3 HBV）into the liver of tree shrews. Methods Serum and liver tissues were collected at indicated times after i. v. injection of rAAV8-1.3 HBV into the tree shrews. The HBsAg, BeAg, HBsAb, HBeAb, HBcAb, ALT and HBV virus load were examined by ELISA and real-time PCR, respectively. The expression of HBcAg and pathological changes in the liver were also observed after the rAAV8-1.3 HBV infection. Results Markers of serum HBV were all positive 2 weeks after and HBcAg-positive hepatocytes were even detected in the liver 55 days after rAAV8-1.3 HBV injection. The copies of HBV DNA in liver reached 104 - 105 at 55 days after rAAVS-1.3HBV injection. Serum HBV DNA could be detected for over one month. Mild pathological changes with elevated ALT were observed after rAAV8-1.3 HBV injection. Conclusions A tree shrew model of acute HBV infection has been successfully established with the recombinant adeno-assoeiated virus 8 carrying 1.3 copies of HBV genome （rAAV8-1.3 HBV）. Our study provides a novel and useful small animal model for the research of immuno- pathologic mechanism as well as the evaluation of vaccine and anti-HBV agents.