CTP-HBcAg_(18-27)-Tapasin诱导C57BL/6小鼠HBV特异性CTL反应的机制  

Mechanism for cytoplasmic transduction peptide-HBcAg_(18-27)-Tapasin to induce HBV specific CTL response in C57BL/6 mice

作　　者：吴姗姗[1] 唐余燕[1] 王洁玲[1] 陈小华[1] 张毅[1] 汤正好[1] 臧国庆[1] 余永胜[1] 

机构地区：[1]上海交通大学附属第六人民医院感染病科,上海市200233

出　　处：《世界华人消化杂志》2016年第17期2688-2695,共8页World Chinese Journal of Digestology

基　　金：上海教育委员会科研创新基金资助项目;No.15ZZ013~~

摘　　要：目的:观察融合蛋白胞质转导肽(cytoplasmic transduction peptide,CTP)-HBcAg_(18-27)-Tapasin通过介导JAK/STAT通路诱导近交系C57BL/6小鼠HBV特异性细胞毒性T淋巴细胞(cytotoxic T lymphocyte,CTL)反应.方法:C57BL/6小鼠随机分为4组:CTPHBcAg_(18-27)-Tapasin实验组、CTP-HBcAg_(18-27)-Tapasin+AG490对照组、AG490对照组、PBS空白组.融合蛋白经肌肉注射免疫小鼠,腹腔注射AG490阻断JAK/STAT通路,CCK-8比色法检测T淋巴细胞增殖活性;流式细胞术检测T淋巴细胞内的的细胞因子;ELISA检测T淋巴细胞分泌细胞因子,Real-time PCR检测JAK/STAT通路信号分子表达水平.结果:CTP-HBcAg_(18-27)-Tapasin组相比CTPHBcAg_(18-27)-Tapasin+AG490组CD8^+IFN-γ^+T双阳性细胞百分比显著增加(P<0.01),淋巴细胞增殖活性差异具有显著性(P<0.01),Th1型细胞因子分泌水平显著增加(P<0.01),其他各组之间没有显著性差异,JAK/STAT信号通路信号中Jak2,STAT4 mRNA分子表达水平CTP-HBcAg_(18-27)-Tapasin组相比其他对照组表达水平差异具有显著性(P<0.05),CTP-HBcAg_(18-27)-Tapasin组Tyk2、STAT1mRNA的分子表达水平相比其他对照组表达水平具有显著性差异(P<0.01).结论:本研究表明CTP-HBcAg_(18-27)-Tapasin通过JAK/STAT信号通路促进Th1型细胞因子的分泌而促进特异性CTL反应.AIM： To observe whether the fusion protein cytoplasmic transduction peptide （CTP） -HBcAg18-27-Tapasin can induce HBV-specific cytotoxic T lymphocyte （CTL） response in C57BL/6 mice via the JAK / STAT pathway.METHODS： C57BL/6 mice were randomly divided into four groups： CTP-HBcAg18- 27- Tapasin alone group, CTP-HBcAg18-27-Tapasin ＋ AG490 group, AG490 alone group, and PBS group. Mice were immunized with the fusion protein through intramuscular injection, and AG490 was administered through intraperitoneal injection. The proliferation of T lymphocytes was observed using CCK-8 assay. The levels of the cytokines secreted by T lymphocytes were detected by ELISA, and the levels of intracellular cytokines of proliferative T lymphocytes were detected by flow cytometry. Expression levels of molecules of the JAK/STAT signal pathway were detected by real-time PCR.RESULTS： The percentage of CD8＋IFN-γ＋ T cells, T lymphocytes proliferative activity and the levels of Thl cytokines in the CTP-HBcAgls_27- Tapasin alone group were significantly increased compared with the CTP-HBcAg18-27-Tapasin ＋ AG490 group （P 〈 0.01）, although there was no statistical significance between the other groups. The expression levels of Jak2 and STAT4 were significantly higher in the CTP-HBcAg18-27-Tapasin alone group than the rest groups （P 〈 0.05）, and the expression levels of Tyk2 and STAT1 were also dramatically increased in the CTP-I-IBcag18-27- Tapasin alone group compared to other groups （P 〈 0.01）.CONCLUSION： CTP-HBcAgls.27-Tapasin fusion protein increases HBV-specific CTL response via the JAK/STAT signal pathway in C57BL/6 mice.

关 键 词：CTP-HBcAg_(18-27)-Tapasin HBV特异性CTL反应 JAK/STAT信号通路 

分 类 号：R512.62[医药卫生—内科学]