胞质转导肽-HBcAg18-27-Tapasin通过激活PI3K/Akt通路抑制HBV转基因小鼠特异性CTL凋亡  

CTP-HBcAg18-27-Tapasin inhibits apoptosis of specific CTL through regulating PI3K/Akt signaling pathway in HBV transgenic mice

作　　者：唐余燕[1] 陈小华[1] 卓萌[1] 宋琳琳[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科,上海200233

出　　处：《胃肠病学和肝病学杂志》2014年第4期420-424,共5页Chinese Journal of Gastroenterology and Hepatology

基　　金：国家自然科学基金青年项目(31000414)

摘　　要：目的探讨胞质转导肽(CTP)-HBcAg18-27-Tapasin融合蛋白通过激活PI3K/Akt信号通路抑制HBV转基因小鼠特异性细胞毒性T淋巴细胞(cytotoxic T cell,CTL)凋亡。方法 HBV转基因小鼠随机分组,100μg CTP-HBcAg18-27-Tapasin、50μg CTPHBcAg18-27-Tapasin、50μg CTP-HBcAg18-27及生理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胞内细胞因子水平及细胞凋亡率,Western blotting检测凋亡相关蛋白PI3K、P-Akt及P-mTOR表达的变化。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(2.74±0.20)%,高于对照组50μg CTP-HBcAg-18-27-Tapasin(2.42±0.53)%及50μg CTP-HBcAg18-27(1.70±0.56)%和空白组(0.66±0.71)%(F=741.45,P=0.000);同时,100μg CTPHBcAg18-27-Tapasin融合蛋白有效抑制特异性CTL凋亡(6.29±0.02)%,低于对照组50μg CTP-HBcAg18-27-Tapasin(7.72±0.15)%及50μg CTP-HBcAg18-27(26.30±1.13)%和空白组(58.26±0.23)%(F=5313,P=0.000),并伴随凋亡相关蛋白PI3K、P-Akt及P-mTOR上调(F=52.61、18.32和94.18,P<0.05)。结论 CTP-HBcAg18-27-Tapasin能有效诱导HBV转基因小鼠特异性CTL的表达,抑制其凋亡,机制与PI3K/Akt通路的激活有关。Objective To investigate the fusion protein cytoplasmic transduction peptide （CTP）-HBcAg18-27- Tapasin inhibited apoptosis of specific cytotoxic T-lymphocytes in HBV transgenic mice by activating the PI3K/Akt sig- naling pathway. Methods Mice were immunized intramuscularly in the left tibialis anterior muscle three times at 1- week intervals with CTP-HBcAgl 8-27-Tapasin （ 100 μg）, CTP-HBcAgl 8-27-Tapasin （ 50 μg）, CTP-HBcAgl 8- 27 （ 50 μg） and PBS. Mice were sacrificed, and splenocytes were collected at day 7 after the third immunization. Intracellular cytokine of proliferative splenocytes and CTL apoptotic rate were analyzed by flow cytometry. The apoptosis related pro- teins levels of PI3K, P-Akt and P-mTOR were examined by Western blotting. Results The number of CTLs induced by 100 μg CTP-HBcAg18-27-Tapasin （2.74 ± 0.20） % and 50 txg CTP-HBcAg18-27-Tapasin （2.42 ± 0.53 ）% were sig- nificantly higher than that in the control group （ 1.70 ＋ 0.56） % and PBS group （0.66 ±0.71 ） % （ F = 741.45, P = 0. 000）. Meanwhile, significantly lower percentages of apoptosis of CD8 ＋ T cells were observed in mice immunized with 100 μg CTP-HBcAg18-27-Tapasin （6.29 ＋ 0.02）% than 50 μg CTP-HBcAg18-27-Tapasin （7.72 ± 0. 15 ）% and 50 μg CTP-HBcAg18-27 （26.30 ±1.13）% and the PBS group （58.26 ±0.23）% （F=5313, P =0.000）. Moreover, the expression of PI3K, P-Akt and P-mTOR were significantly upregulated in CTP-HBcAg18-27-Tapasin group compared with control group （F = 52.61, 18.32 and 94. 18, P 〈 0.05）. Conclusion The fusion protein CTP-HBcAg18-27- Tapasin could inhibit apoptosis of specific cytotoxic T-lymphocytes in HBV transgenic mice by activating the PI3K/Akt signaling pathway.

关 键 词：胞质转导肽-HBcAg18-27-Tapasin P13K Akt通路 细胞毒T淋巴细胞 凋亡 乙肝病毒 

分 类 号：R512.62[医药卫生—内科学]