急性乙型肝炎病毒感染小鼠模型的建立

Development of the mouse model for acute hepatitis B virus infection

目的采用尾静脉液压法建立小鼠急性HBV感染的动物模型。方法以液压法将具有复制能力的HBV质粒pAAV-HBV1.2通过尾静脉注射到免疫功能正常的BALB/c小鼠体内,注射后第1、2、4、6、8d,分别采用改良赖氏法、时间分辨免疫荧光法、实时荧光定量PCR检测小鼠血清中谷丙转氨酶(ALT)、HBsAg、HBeAg、抗HBs、抗HBe、HBV DNA的水平,免疫组化检测肝组织HBsAg、HBcAg的表达。结果16只小鼠注射pAAV-HBV1.2后,有14只(85.7%)小鼠在注射后第1d血清中可检测到HBsAg,小鼠血清中HBsAg和HBeAg水平在第1d达高峰,之后逐渐下降,第8d均未能检测到。小鼠血清中HBV DNA在第2d达高峰,之后仍维持在较高水平,至第8d时为1.9×104copies/mL。至第8d肝组织中可见约5%的HBcAg阳性肝细胞和2%的HBsAg阳性肝细胞。结论采用尾静脉液压法成功的建立了小鼠急性HBV感染的动物模型。

A mouse model for acute hepatitis B virus （HBV） infection was established by using the hydrodynamical in jection of mouse tail vein, in which the immunocompetent BALB/c mice were hydrodynamically injected with a competent replication plasmid pAAV-HBV1.2 having 1.2 fold ovevlength of HBV DNA. On day 1, 2, 4, 6 and 8 after injection, the levels of HBsAg, HBeAg and HBV DNA in blood serum were detected by using ELISA and fluorogenic quantitative PCR assay （FQPCR）. And on day 8. HBsAg and HBeAg in liver tissue were assayed by immunohistoehemical staining. It was found that HBsAg in blood serum could be detected on day 1 after infection in 14 of 16 mice （85.7%） injected with pAVV-HBV1.2 by using ELISA assay and the peak levels of HBsAg and HBeAg were attained during the first day after injection and then it dropped down gradually up to day 8 following injection. The titer of HBV DNA in blood serum attained its peak on day 2 and maintained a high level later on. On day 8 after injection, its titer was 1.9 × 10^4 copies/mL. The percentage of HBcAg-positive hepatocytes and HBsAg-positive hepatocytes in liver tissues were 5 % and 2%respectively. Thus, by using the hydrodynamic injection with the competent replication plasmid, a mouse model for acute HBV infection is successively developed.