不同种小鼠乙型肝炎模型的建立及其感染状况研究

Establishment of hepatitis B virus infection model in different mice by hydrodynamic injection of pAAV/HBV1.2 plasmid

目的探讨建立急性或慢性乙型肝炎病毒(HBV)感染小鼠模型的方法。方法采用高压尾静脉注射p AAV/HBV1.2表达质粒,建立急性或慢性HBV感染小鼠模型,采用ELISA法检测HBV抗原,采用RT-PCR法检测免疫相关分子基因,使用FACS检测肝内淋巴细胞活化情况,采用酶联免疫斑点实验(ELISPOT)法检测脾脏相关免疫细胞特征。结果成功建立急性和慢性高压尾静脉注射HBV感染小鼠模型;在急性BALB/c小鼠模型,血清HBs Ag持续时间分别为(3.25±1.04)w,显著短于慢性C57BL/6小鼠组【(10.0±3.74)w,P<0.05)】;在BALB/c小鼠模型,注射2.5μg病毒质粒小鼠血清HBs Ag持续时间为(3.67±1.03)w,显著长于注射50μg组【(2.33±0.52)w,(P<0.05)】;在高压尾静脉注射后第10 d,BALB/c小鼠脾脏出现了HBc Ag特异性T淋巴细胞。结论成功建立高压尾静脉注射HBV感染小鼠模型,发现宿主遗传背景、免疫应答状态和病毒剂量影响HBV感染小鼠模型的建立。

Objective To establish the acute or chronic HBV-infeetion mouse model. Methods Acute or chronic HBV-infection mouse model was established by hydrodynamic injection of pAAV/HBV1.2 plasmid. Serum HBsAg were detected by ELISA. The immune-related molecule mRNA were detected by real-time PCR. The activated lymphocytes in the livers were detected by FACS and ELISPOT was used to detect the HBV-specific cellular immune response. Results We successfully established the acute and chronic HBV-infection mouse model by hydrodynamic injection of pAAV/HBV1.2 plasmid;The duration of serum HBsAg secretion in acute HBV-infected BALB/c mice was （3.25±1.04） weeks,significantly shorter than that in chronic HBV-infected C57BL/6 mice [（10.0±3.74） weeks,P〈0.05l;The duration of serum HBsAg secretion in BALB/c mice injected with low dose （2.5μg） of pAAV/HBV1.2 plasmid was （3.67±1.03） weeks,significantly longer than that in high dose （50μg） group I-（2.33±0.52） weeks,P〈0.05];On the tenth day after hydrodynamic injection,HBcAg special T lymphocytes appeared in the spleen of BALB/c mice. Conclusion HBV-infection mouse model is successfully established by hydrodynamic injection of pAAV/HBV1.2 plasmid and we find that mouse genetic background, immune response and viral doses will impact the establishment of this kind of HBV-infection mouse model.