实用肝脏病杂志 ›› 2017, Vol. 20 ›› Issue (5): 533-537.doi: 10.3969/j.issn.1672-5069.2017.05.007

• 实验性肝炎 • 上一篇    下一篇

不同种小鼠乙型肝炎模型的建立及其感染状况研究

周云,李盛,唐宗生,杨东亮,宋景娇   

  1. 430022 武汉市 华中科技大学同济医学院附属协和医院感染性疾病科(周云,李盛,唐宗生,杨东亮);附属同济医院实验医学中心(宋景娇);河南大学医学院(周云)
  • 收稿日期:2016-10-26 出版日期:2017-10-10 发布日期:2017-10-17
  • 通讯作者: 宋景娇,E-mail: jjsong@tjh.timu.edu.cn
  • 作者简介:周云,男,36岁,医学博士。主要从事感染与免疫研究。E-mail: zyky120@126.com
  • 基金资助:
    国家传染病防治科技重大专项项目(编号:2008ZX10002-011)

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

Zhou Yun,Li Sheng,Tang Zongsheng,et al.   

  1. Department of Infectious Disease,Union Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430022,China
  • Received:2016-10-26 Online:2017-10-10 Published:2017-10-17

摘要: 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pAAV/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HBsAg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2.5 μg病毒质粒小鼠血清HBsAg持续时间为(3.67±1.03) w,显著长于注射50 μg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HBcAg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。

关键词: 乙型肝炎, pAAV/HBV1.2表达质粒, 高压尾静脉注射, 小鼠

Abstract: Objective To establish the acute or chronic HBV-infection 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<0.05];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[(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.

Key words: Hepatitis B, pAAV/HBV1.2 plasmid, Hydrodynamic injection, Mice