干扰素α调控乙型肝炎病毒特异性T细胞免疫应答抑制慢性感染小鼠模型体内病毒基因表达  

Interferon-α modulates HBV specific T-cell response to inhibit viral gene expression in chronic HBV infection mouse model

作　　者：刘娜[1] 葛军[1] 沈思岚 许燕妮 张小勇[1] 刘红艳[1] LIU Na;GE Jun;SHEN Silan;XU Yanni;ZHANG Xiaoyong;LIU Hongyan(Hepatology Unit and Department of Infectious Diseases,Nanfang Hospital,Southern Medical University,Guangzhou 510515,China)

机构地区：[1]南方医科大学南方医院(第一临床医学院)感染内科,广州510515

出　　处：《中国感染与化疗杂志》2018年第2期177-183,共7页Chinese Journal of Infection and Chemotherapy

基　　金：广东省大学生创新创业训练计划项目(201512121117);广东省自然科学基金(2014A030313299)

摘　　要：目的在慢性乙型肝炎病毒(HBV)感染小鼠模型中,探讨干扰素α(IFN-α)抑制HBV基因表达的效应及其机制。方法 C57BL/6j小鼠采用高压尾静脉注射p AAV-HBV1.2质粒建立慢性HBV感染小鼠模型,分组同时注射IFN-α-2a表达质粒p KCMvint.IFN-α-2a和对照质粒p KCMvint后,采用酶联免疫吸附测定(ELISA)检测小鼠体内IFN-α表达,雅培ARCHITECT i2000SR检测血清HBs Ag、HBe Ag水平,锥虫蓝染色计数检测肝脾内总淋巴细胞频数,流式细胞术检测CD8^+T细胞的频数、HBV特异性CD8^+T细胞的频数及功能。结果干扰素质粒组IFN-α表达明显高于对照组(P<0.01),且血清HBs Ag在注射后第12天就有明显下降,显著快于对照组,并能完全清除HBe Ag;干扰素质粒组的肝内总淋巴细胞、CD8^+T细胞、HBV特异性CD8^+T细胞频数均有高于对照组趋势,且CD8^+T细胞频数两组差异有统计学意义(P<0.05);干扰素质粒组HBV特异性CD8^+T细胞分泌IFN-γ、TNF-α和IL-2能力均有高于对照组的趋势,且分泌IFN-γ能力两组差异具有统计学意义(P<0.05)。结论 IFN-α体内过表达可以上调小鼠肝内HBV特异性CD8^+T细胞的频数和功能,抑制慢性HBV感染小鼠模型内HBs Ag和HBe Ag的表达。提示IFN-α可以通过增强抗HBV免疫应答来发挥抗病毒效应。Objective To explore the antiviral effect and mechanism of interferon-α(IFN-α)in chronic HBV infection mouse model by hydrodynamic tail vein injection.Methods Chronic HBV infection mouse model was constructed with C57BL/6j mice by hydrodynamic tail vein injection of pAAV-HBV1.2 plasmid.Serum IFN-αwas determined by ELISA after injection of IFNα-expressing plasmid(i.e.,plasmid pKCMvint.IFN-α-2a)or control plasmid pKCMvint.HBsAg and HBeAg levels were assayed on Abbot ARCHITECT i2000SR.Total lymphocytes in liver or spleen were counted and the frequency and function of CD8+T cells and HBV-specific CD8+T cells were analyzed by flow cytometry.Results Serum IFN-αexpression level was significantly higher in the animals injected with pKCMvint.IFN-α-2a plasmid than in control group(P<0.01).Serum HBsAg decreased quickly 12 days after injection and significantly lower than control group.Serum HBeAg was undetectable after IFN-αexpression.Interestingly,the frequency of CD8+T cells in the liver of animals injected with pKCMvint.IFN-α-2a plasmid was significant higher than control group(P<0.05),while total liver lymphocytes and HBV-specific CD8+T cells had a similar trend.Consistently,the HBV-specific CD8+T cells in IFN-α-expressing animals showed higher level of producing IFN-γ,TNF-αand IL-2 than control group.IFN-γproduction was significantly different between IFN-α-expressing group and control group(P<0.05).Conclusions IFN-αcan increase the frequency and function of liver CD8+T cells to inhibit HBV gene expression in chronic HBV infection mouse model.

关 键 词：干扰素Α 慢性乙型肝炎 乙型肝炎病毒 T细胞应答 适应性免疫 

分 类 号：R512.62[医药卫生—内科学]