自然杀伤细胞分泌干扰素γ促进乙肝病毒的清除  

Natural killer cell promotes the HBV clearance via secreting IFN-γ

作　　者：胡辛[1] 胡春松[1] 黄保军[1] 

机构地区：[1]安徽医科大学基础医学院免疫教研室,合肥230032

出　　处：《安徽医科大学学报》2017年第2期215-220,共6页Acta Universitatis Medicinalis Anhui

基　　金：安徽省自然科学基金(编号:1208085MH134)

摘　　要：目的探讨自然杀伤细胞(NK)在乙型肝炎病毒(HBV)清除过程中的作用机制。方法通过对成年小鼠高压注射20μg p GEM4Z/HBV1.2质粒建立HBV急性排斥小鼠模型,采用放射免疫试剂盒定量检测不同时间点(1、2、3、4、5周)小鼠血清中HBsAg、免疫组化检测肝脏组织中HBc Ag来评估模型的建立。流式细胞术分析对照小鼠和HBV小鼠肝脏中NK细胞比例和绝对数量变化,进一步分析两种小鼠NK细胞活化情况和分泌干扰素(IFN)-γ的能力变化。通过肝脏组织切片HE染色以及谷丙转氨酶检测判断小鼠肝脏损伤状况。最后,通过PK136抗体清除小鼠NK细胞以及IFN-γ中和抗体进一步证实NK细胞分泌的IFN-γ是否参与HBV的清除过程。结果小鼠高压注射20μg p GEM4Z/HBV1.2质粒后,第1周血清中HBsAg和肝脏组织中HBc Ag表达量都较高,但呈降低趋势,在4~5周后几乎转阴,很好地模拟了临床上急性感染HBV的病例。与对照组相比,HBV小鼠肝脏中NK细胞比例和绝对数量明显增高;进一步研究显示HBV小鼠NK细胞活化分子CD69表达也显著上升,同时IFN-γ分泌增加。与此同时,小鼠肝脏呈无损伤状态。预先清除NK细胞或阻断IFN-γ的功能能显著增加HBV小鼠中相关抗原的含量,延缓HBV的排斥。结论在HBV急性排斥模型中,NK细胞通过分泌IFN-γ促进HBV的清除。Objective To investigate the mechanism of NK cell in eliminating the Hepatitis B virus during HBV in- fection. Methods Acute HBV infection model was established by injecting adult mice hydrodynamically with 20μg of pGEM4Z/HBV1.2 plasmid. This model was evaluated by detecting serum level of HBsAg and HBcAg in liver tis- sue at the indicated time points by radioimmunoassay and immunohistochemistry respectively. The variation of frequency and absolute number of NK cell was analyzed between wide type （WT） mice and HBV plasmid-injected mice. Furthermore, the activation and the IFN-γ production of NK cell Tere investigated in these mice by flow cytometry. HE staining and alanine transaminase（ALT） dectection were used to observe liver injury. To test whether NK cell and IFN-γ were involved in HBV elimination, we used PK136 antibody to clear NK cell and IFN-5, neutral- ization antibody toblock IFN-γ effect. Results After the hydrodynamic injection with 20μg of pGEM4Z/HBV1.2, the serum level of HBsAg and expression of HBcAg in liver tissue were very high at 1 week, but then decreased gradually. However, these antigens almost became negative at 4 to 5 weeks, which mimic acute HBV infection patients. Compared with NK cell from WT mice, the frequency and absolute number of NK cell increased significantly from HBV mice. Also, the NK cells express higher level of CD69 and produce more IFN-γ. Meanwhile, there was no liver injury in HBV mice. Depletion of NK cell or blocking IFN-γ effect in HBV mice could significantly in- crease the level of HBV related antigens. Conclusion In the mouse model of acute HBV infection, NK cell could promote the HBV elimination through secreting IFN-γ.

关 键 词：HBV NK细胞 IFN-Γ 

分 类 号：R392.12[医药卫生—免疫学]