补肾解毒方阻断程序性死亡受体1/程序性死亡配体1信号通路对辅助慢性乙型肝炎树突状细胞疫苗抗乙型肝炎病毒的免疫效果与机制研究  

Effect and mechanism study of Bushen Jiedu prescription blocking programmed death receptor 1/programmed death ligand 1 pathway on anti-hepatitis B virus immunity of dendritic cell vaccine assisted by chronic hepatitis B

作　　者：蔡本强[1] 贺劲松[1] 邢宇锋[1] CAI Benqiang;HE Jinsong;XING Yufeng(Department of Liver Disease,Shenzhen Hospital of Traditional Chinese Medicine,Shenzhen,Guangdong 518033)

机构地区：[1]广东省深圳市中医院肝病科

出　　处：《河北中医》2019年第9期1391-1396,共6页Hebei Journal of Traditional Chinese Medicine

基　　金：深圳市科技工贸和信息化委员会2011年深圳市科技计划项目(医疗卫生类)(编号:201102196)

摘　　要：目的研究补肾解毒方阻断程序性死亡受体1(PD-1)/程序性死亡配体1(PD-L1)信号通路对辅助慢性乙型肝炎(以下简称乙肝)树突状细胞(DC)疫苗抗乙肝病毒(HBV)的免疫效果与机制。方法将40只BALB/c HBV转基因小鼠按照随机数字表法分为补肾解毒方组、PD-L1抗体组、磷酸缓冲液(PBS)组及联合组4组,每组10只。4组分别予补肾解毒方、PD-L1单克隆抗体、PBS溶液及补肾解毒方联合PD-L1单克隆抗体,持续给药2周后处死小鼠,观察小鼠脾脏组织γ干扰素(IFN-γ)水平、CD8^+T淋巴细胞的PD-1表达情况及乙肝表面抗原(HBsAg)特异性细胞毒性T淋巴细胞(CTL)的杀伤活性。结果联合组小鼠脾脏组织IFN-γ水平高于补肾解毒方组、PD-L1抗体组、PBS组(P<0.05);补肾解毒方组、PD-L1抗体组小鼠脾脏组织IFN-γ水平高于PBS组(P<0.05);补肾解毒方组与PD-L1抗体组小鼠脾脏组织IFN-γ水平比较差异无统计学意义(P>0.05)。联合组小鼠脾脏组织CD8^+T淋巴细胞PD-1表达均低于补肾解毒方组、PD-L1抗体组、PBS组(P<0.05);补肾解毒方组、PD-L1抗体组CD8^+T淋巴细胞PD-1表达均低于PBS组(P<0.05);PD-L1抗体组CD8^+T淋巴细胞PD-1表达低于补肾解毒方组(P<0.05)。当效靶比为100∶1、50∶1时,联合组小鼠脾脏组织HBsAg特异性CTL杀伤活性均高于补肾解毒方组、PD-L1抗体组、PBS组(P<0.05);PD-L1抗体组HBsAg特异性CTL杀伤活性均高于补肾解毒方组、PBS组(P<0.05);补肾解毒方组与PBS组HBsAg特异性CTL杀伤活性比较差异无统计学意义(P>0.05)。当效靶比为25∶1时,补肾解毒方组、PD-L1抗体组、联合组小鼠脾脏HBsAg特异性CTL杀伤活性均高于PBS组(P<0.05);补肾解毒方组、PD-L1抗体组、联合组小鼠脾脏HBsAg特异性CTL杀伤活性两两比较差异均无统计学意义(P>0.05)。结论 HBV感染时阻断PD-1/PD-L1信号通路可起到恢复特异性T淋巴细胞功能的作用,补肾解毒方能有效发挥对HBV感染时的T淋巴细胞Objective To study the effect and mechanism of Bushen Jiedu prescription blocking the programmed death receptor 1(PD-1)/programmed death ligand 1(PD-L1) pathway on the anti-hepatitis B virus(HBV) immunity of dendritic cell(DC) vaccine assisted by chronic hepatitis B.Methods 40 BALB/c HBV transgenic mice were randomly divided into four groups: Bushen Jiedu prescriptio group, PD-L1 antibody group, phosphate buffer solution(PBS) group and combination group, with 10 rats in each group. Four groups were respectively given Bushen Jiedu prescription, PD-L1 monoclonal antibody, PBS solution and Bushen Jiedu prescription combined with PD-L1 monoclonal antibody. Mice were killed after 2 weeks of continuous administration. The levels of interferon γ(IFN-γ) in spleen, the expression of PD-1 in CD8^+ T lymphocytes and the killing activity of hepatitis B surface antigen(HBsAg) specific cytotoxic T lymphocytes(CTL) were observed. Results The level of IFN-γ in the spleen tissue of mice in the combined group were higher than those in the Bushen Jiedu prescription group, the PD-L1 antibody group and the PBS group(P<0.05). The level of IFN-γ in the Bushen Jiedu prescription group and the PD-L1 antibody group were higher than those in the PBS group(P<0.05). There was no significant difference in the levels of IFN-γ between the Bushen Jiedu prescription group and the PD-L1 antibody group(P>0.05). The expression of PD-1 on CD8^+T lymphocyte in the combined group was lower than that in the Bushen Jiedu prescription group, the PD-L1 antibody group and the PBS group(P<0.05), and the expression of PD-1 on CD8^+T lymphocyte in the Bushen Jiedu prescription group and the PD-L1 antibody group was lower than that in the PBS group(P<0.05). The expression of PD-1 on CD8^+T lymphocyte in PD-L1 antibody group was lower than that in Bushen Jiedu prescription group(P<0.05). When the effective-target ratio was 100 ∶1 and the effective-target ratio was 50 ∶1, the killing activity of HbsAg CTL in spleen tissue of mice in combined group was hi

关 键 词：乙型肝炎 乙型肝炎病毒 中药疗法 疫苗 PD-1/PD-L1通路 小鼠 动物实验 

分 类 号：R512.62[医药卫生—内科学] R373.21[医药卫生—临床医学]