CTP-HBcAg_(18-27)-Tapasin联合佐剂CpG ODN免疫转基因小鼠的免疫应答研究  

Immune response elicited in HBV-transgenic mice by cytoplasmic transduction peptide-HBcAg_(18-27)-Tapasin adjuvanted with CpG ODN

作　　者：吴姗姗[1] 唐余燕[1] 陈小华[1] 张毅[1] 王洁玲[1] 汤正好[1] 臧国庆[1] 余永胜[1] WU Shanshan;TANG Yuyan;CHEN Xiaohua;ZHANG Yi;WANG Jieling;TANG Zhenghao;ZANG Guoqing;YU Yongsheng(Department of Infectious Diseases, Shanghai Jiao Tong University Affiliated Sixth People＇s Hospital, Shanghai 200233, Chin)

机构地区：[1]上海交通大学附属第六人民医院感染病科,上海200233

出　　处：《中国临床药理学与治疗学》2018年第3期271-276,共6页Chinese Journal of Clinical Pharmacology and Therapeutics

基　　金：上海教育委员会科研创新项目(15ZZ013);上海市卫生和计划生育委员会(201640283)

摘　　要：目的:观察以非甲基化寡聚脱氧核苷酸基序(CpG oligodeoxynucleotides,CpG ODN)为佐剂联合融合蛋白胞质转导肽(CTP)-HBc Ag18-27-Tapasin免疫乙型病毒性肝炎(hepatitis B virus,HBV)转基因小鼠的免疫反应。方法:HBV转基因小鼠随机分为5组,实验组:CTP-HBc Ag18-27-Tapasin+CpG ODN组,对照组:CTP-HBc Ag18-27-Tapasin,干扰素(IFN-α)组,CpG ODN组,空白组为生理盐水组。融合蛋白及CpG ODN经肌肉注射免疫小鼠,流式细胞仪检测病毒特异性T细胞反应(HBV-specific cytotoxic T cell,CTL)变化,全自动免疫分析仪检测血清乙肝表面抗原(hepatitis B virus surface antigen,HBs Ag)水平,ELISA检测T淋巴细胞培养上清白介素(IL)-2、γ-干扰素(IFN-γ)水平,实时荧光定量PCR(real time quantitative polymerase chain reaction,q RT-PCR)检测HBV DNA水平,免疫组化观察HBV转基因小鼠肝脏中HBs Ag水平。结果:CTP-HBc Ag18-27-Tapasin+CpG ODN组中IFN-γ+CD8+双阳性T细胞百分比增高,细胞培养上清IL-2、IFN-γ水平升高,肝脏病理显示炎性细胞增多,免疫组化显示HBs Ag表达水平明显下降,同时对血清HBs Ag及HBV DNA水平进行比较,CTP-HBc Ag18-27-Tapasin有明显抑制作用。结论:CpG-ODN作为佐剂可以增强融合蛋白CTP-HBc Ag18-27-Tapasin诱导的HBV转基因小鼠抗病毒免疫应答。AIM： To investigate the effects of fusion protein of cytoplasmic transduction peptide（ CTP）-HBcAg_（18-27）-Tapasin adjuvanted with CpG ODN on immune response and anti-viral response in HBV transgenic mice. METHODS： HBV-transgenic mice were randomly divided into 5 groups： experimental group： CTP-HBcAg_（18-27）-Tapasin ＋ CpG ODN,control group： CpG ODN,IFN-α and CTPHBcAg_（18-27）-Tapasin,vehicle group： normal saline.Serum samples were collected at various times before and after immunization. Intracellular cytokine in T lymphocytes was analyzed by flow cytometry,and the levels of IL-2 and IFN-γ secreted by T lymphocytes were detected by ELISA. The expression of HBs Ag in liver sections was detected by immunohistochemistry. Sera levels of HBV DNA and HBs Ag were examined by q RT-PCR and automatic enzymelinked immunosorbent assay, respectively. RESULTS： An increased percentage of IFN-γ＋CD8＋T cells was detected in CTP-HBcAg_（18-27）-Tapasin ＋ CpG ODN group and predominantly Th1 cellular immune response. In addition,the expression levels of serum HBs Ag and HBV DNA were significantly inhibited,along with evident down-regulation of HBs Ag in liver samples than CTP-HBcAg_（18-27）-Tapasin alone group.CONCLUSION： CpG ODN acts as adjuvant could enhance anti-virus immune responses in HBV transgenic mice immunized with fusion protein CTP-HBcAg_（18-27）-Tapasin.

关 键 词：胞质转导肽 抗病毒反应 佐剂 

分 类 号：R965.2[医药卫生—药理学]