摘要
目的 探讨经分子伴侣Tapasin修饰抗原肽HBcAg18-27经CTP转导体内诱导特异性细胞毒性T淋巴细胞(CTL)对HBV转基因小鼠病毒的抑制作用. 方法 20只HBV转基因小鼠随机分组,100 μg CTP-HBcAg18-27-Tapasin,50μg CTP-HBcAg18-27-Tapasi,50 μg CTP-HBcAg18-27及等渗盐水组经肌肉免疫小鼠,每周一次,共3次.流式细胞仪检测脾细胞中CTL水平,微粒子酶免疫分析法(MEIA)检测血清中HBsAg水平,荧光定量聚合酶链反应(PCR)检测HBVDNA水平,肝脏HE染色及免疫组织化学方法检测HBsAg表达.数据用均数±标准差((x-)±s)表示,用SPSS16.0软件进行单因素方差分析(one-way ANOVA),组间两两比较采用LSD法. 结果 100μg CTP-HBcAg18-27-Tapasin融合蛋白免疫转基因小鼠后,能有效上调特异性CTL数量(2.70％±0.20％),其水平高于对照组50 μg CTP-HBcAg18-27-Tapasin (1.66％±0.53％)及50μg CTP-HBcAg18-27 (1.26％±0.56％)和空白组(0.75％±0.71％),F=741.45,P=0.000.肝脏HE染色及免疫组织化学结果显示实验组肝组织中炎性细胞的数量明显增多及HBsAg的表达显著减少,同时对小鼠血清中HBsAg及HBV DNA有明显的抑制作用. 结论 分子伴侣Tapasin修饰抗原肽HBcAg 18-27经CTP转导免疫HBV转基因小鼠后能增加特异性CTL数量,显著降低血清中HBsAg及HBV DNA水平,同时抑制肝脏中HBsAg的表达.
Objective To investigate the effect of protein transduction domain-hepatitis B virus core antigen (CTP-HBcAg18-27)-Tapasin fusion protein-induced specific cytotoxic T lymphocyte (CTL) response on hepatitis B virus (HBV) replication in HBV transgenic mice.Methods Twenty HBV-transgenic mice were randomly divided into two groups for a 3-week course of once weekly subcutaneous immunizations with either CTP-HBcAg18-27-Tapasin fusion protein or CTP-HBcAg18-27.Mice administered isotonic saline served as blank controls.Expressions of cytokines in splenocytes were analyzed by flow cytometry.Serum levels of hepatitis B surface antigen (HBsAg) and HBV DNA were determined by microparticle enzyme immunoassay and real-time fluorescent PCR assay,respectively.Expression of HBsAg in hepatic tissues was detected by immunohistochemistry.Results Immunization with 100 μg of CTP-HBcAg18-27-Tapasin fusion protein led to a significant increase in proportions of CTLs in spleen (2.70％ ± 0.20％ vs.50 μg of CTP-HBcAg18-27-Tapasin:1.66％ ± 0.53％,50 μg of CTP-HBcAg18-27:1.26％ ± 0.56％,and blank controls:0.75％ ± 0.71％;F=741.45,P =0.000) and up-regulation of inflammatory cells in hepatic tissue.In addition,both immunizations of CTP-HBcAgl 8-27-Tapasin led to significant decreases in serum HBsAg and HBV DNA levels compared to those in the CTP-HBcAg18-27 group.Conclusion HBV-related modification of the expression of the molecular chaperone Tapasin may affect its interaction with intracellular antigen peptides,thereby leading to increases the number of specific CTLs in the spleen,decreases in serum HBsAg and HBV DNA levels,and down-regulation of HBsAg expression in hepatic tissue.These results obtained in HBV-transgenic mice suggest that the CTP-HBcAgl 8-27-Tapasin fusion protein has anti-HBV activity.
出处
《中华肝脏病杂志》
CAS
CSCD
北大核心
2014年第1期-,共6页
Chinese Journal of Hepatology
基金
国家自然科学基金
