沉默PD-1基因对AAV8介导的HBV感染大鼠T细胞免疫应答的影响  

Effect of silencing PD-1gene on AAV8-mediated T cell immune response in HBV-infected rats

作　　者：王燕[1] 罗丹[1] 雷敏[1] 胡耀[2] 颜晓玉[3] WANG Yan;LUO Dan;LEI Min;HU Yao;YAN Xiao-yu(Affiliated Hospital of Chengdu University,Chengdu,Sichuan 610081,China)

机构地区：[1]成都大学附属医院血透室,四川成都610081 [2]成都大学附属医院肾内科,四川成都610081 [3]成都大学附属医院呼吸感染科,四川成都610081

出　　处：《中华医院感染学杂志》2021年第17期2584-2588,共5页Chinese Journal of Nosocomiology

摘　　要：目的探究沉默PD-1基因对腺相关病毒(AAV)8介导的乙型肝炎病毒(HBV)感染大鼠T细胞免疫应答的影响。方法选取40只正常大鼠,随机选取10只不做处理,为正常组,其余30只建立AAV8介导的HBV感染大鼠模型,建模成功后,随机选取10只不增加处理,为模型组,将剩余20只模型大鼠分为沉默PD-1组与过表达PD-1组。其中沉默PD-1组进行沉默程序性死亡细胞受体-1(PD-1)干预,过表达PD-1组进行过表达PD-1干预。对大鼠乙型肝炎表面抗原(HBsAg)、乙型肝炎e抗原(HBeAg)水平进行检测与比较,RT-PCR法检测各组大鼠HBV DNA,使用流式细胞仪对CD_(8)^(+)细胞与T细胞表面PD-1分子表达检测及肿瘤坏死因子-α(TNF-α)、干扰素γ(IFN-γ)及Tim3阳性细胞率进行分析。结果与正常组比较,模型组、过表达PD-1组及沉默PD-1组HBsAg、HBeAg水平均较高,与模型组比较,过表达PD-1组HBsAg、HBeAg水平均较高,沉默PD-1组HBsAg、HBeAg水平均较低(P<0.05);与正常组比较,模型组、过表达PD-1组及沉默PD-1组大鼠肝脏IFN-γ、TNF-α阳性细胞率均较低,Tim3阳性细胞率较高,与模型组比较,过表达PD-1组IFN-γ、TNF-α阳性细胞率均较低,Tim3阳性细胞率较高,沉默PD-1组IFN-γ、TNF-α阳性细胞率均较高,Tim3阳性细胞率较低(P<0.05);与正常组比较,模型组、过表达PD-1组及沉默PD-1组大鼠CD_(8)^(+)淋巴细胞占比均较低,T细胞表面PD-1表达较高,与模型组比较,过表达PD-1组CD_(8)^(+)淋巴细胞占比较低,T细胞表面PD-1表达较高,沉默PD-1组CD_(8)^(+)淋巴细胞占比较高,T细胞表面PD-1表达较低(P<0.05)。结论在AAV8介导的HBV感染大鼠中进行沉默PD-1基因干预,能够显著肝纤维化进程,提升T细胞的相关活性,对机体内免疫代谢功能进行调节,增强机体抗病毒能力,改善预后,提升机体免疫应答能力,抑制乙型肝炎病毒进展。OBJECTIVE To explore the effect of silencing PD-1gene on adeno-associated virus(AAV)8-mediated T cell immune response in HBV-infected rats.METHODS A total of 40normal rats were randomly selected,10of which left untreated to be the normal group,and the remaining 30rats were established to AAV8-mediated HBV infection models.After successful modeling,10rats were randomly selected and left untreated to be the model group,and the remaining 20model rats were divided into the silencing PD-1group and the overexpression PDgroup.Among them,the silencing PD-1group underwent the silencing programmed death cell receptor-1(PD-1)intervention,and the overexpressing PD-1group underwent the overexpression PD-1intervention.The levels of HBsAg and HBeAg of the rats were measured and compared,the HBV DNA of the rats from each group was detected by RT-PCR,and the expression of PD-1molecules on surfaces of CD_(8)^(+)cells and T cells were detected by means of flow cytometry,and the positive rates of cells of tumor necrosis factor-α(TNF-α),interferonγ(IFN-γ)and Tim3were observed.RESULTS The levels of HBsAg and HBeAg of the model group,the overexpression PD-1group and the silencing PD-1group were significantly higher than those of the normal group;the levels of HBsAg and HBeAg of the overexpressing PD-1group were significantly higher than those of the model group,while the levels of HBsAg and HBeAg of the silencing PD-1group were significantly lower than those of the model group(P<0.05).The positive rates of cells of IFN-γand TNF-αin livers of the model group,the overexpression PD-1group and the silencing PD-1group were significantly lower than those of the normal group,the positive rate of cells of Tim was higher;the positive rates of cells of IFN-γand TNF-αof the overexpressing PD-1group were significantly lower than those of the model group,the positive rate of cells of Tim3was significantly higher,the positive rates of cells of IFN-γand TNF-αof the silencing PD-1group were significantly higher than those of the model group,

关 键 词：乙型肝炎病毒 免疫应答 程序性死亡细胞受体-1 T细胞 

分 类 号：R512.62[医药卫生—内科学]