补肾解毒方联合程序性死亡受体配体-1抗体对HBV转基因小鼠T细胞免疫应答影响的研究

摘要

目的:观察补肾解毒方联合程序性死亡受体配体-1 (PD-L1)抗体免疫,对HBV转基因小鼠特异性细胞毒性T细胞(CTL)细胞免疫应答的影响.方法:HBV转基因小鼠,使用补肾解毒方灌胃,PD-L1腹腔注射,共两周.检测小鼠脾脏淋巴细胞IFN-γ分泌水平、脾脏CD8+T细胞程序性死亡受体-1(PD-1)表达情况及脾脏HBsAg特异性T淋巴细胞体外杀伤活性.结果:免疫两周后,小鼠脾脏淋巴细胞IFN-γ分泌水平与脾脏HBsAg特异性T淋巴细胞体外杀伤活性,联合组明显高于PD-L1抗体组、补肾解毒方组及磷酸盐缓冲液(PBS)组,统计学分析差异均有显著性意义(P＜0.01或0.05).脾脏CD8+T细胞PD-1表达水平,联合组较PD-L1抗体组、补肾解毒方组及PBS组明显降低,差异均有显著性意义(P＜0.01或0.05).结论:补肾解毒方联合PD-L1抗体免疫,可显著提高HBV转基因小鼠体内HBV抗原特异性CTL活性,提高脾脏淋巴细胞分泌IFN-γ水平,能降低脾脏CD8+T细胞表面PD-1分子的表达.这种联合方法有利于恢复慢性HBV感染时T细胞的免疫功能,增强其免疫应答水平.%Objective:To observe the effect of a Chinese herbal formula (invigorating the kidney and detoxicating decoction,IKDD) combined with programmed death ligand-1 (PD-L1) on the specific cytotoxic T-lymphocyte (CTL) in hepatitis B virus (HBV) transgenic mice.Methods:HBV transgenic mice were intragastric administrated with medication IKDD,combined with intraperitoneal injection of PD-L1 for 2 weeks.Then detected IFN-γlevels in splenic lymphocytes,the expression of PD-1 in splenic CD8 + T cell,and cytotoxic activity of HBsAg specific T lymphocytes in spleen.Results:After dealt with medication IKDD and PD-L1 for 2 weeks,Test results indicated that IFN-γ levels and cytotoxic activity of HBsAg specific T lymphocytes were both higher than that in either PD-L1 antibody group or IKDD group alone and PBS group (P ＜ 0.01 or O.05).In the mean time,compared with either PD-L1 antibody group or IKDD group alone,combining medication IKDD and PD-L1 antibody obviously decreased the expression of PD-1 in splenic CD8 + T cell,there was a statistically significant difference (P ＜ 0.01or 0.05).Conclusion:Combining medication IKDD and PD-L1 antibody administration in HBV transgenic mice could markedly increase IFN-γlevels and cytotoxic activity of HBV antigen specific CTL,while decrease the expression of PD-1 molecule in the surface of splenic CD8 + T cell.The combination is beneficial to recover the immunologic function of T cell and enhance the immune response when chronically infected with hepatitis B virus.