胞质转导肽-HBcAg_(18-27)-Tapasin诱导C57BL/6小鼠T淋巴细胞分泌Th1型细胞因子及HBV特异性CTL的表达

Secretion of Th1-type cytokines and production of HBV-specific cytotoxic T lymphocytes in C57BL/6 mice induced by cytoplasmic transduction peptide-HBcAg_(18-27)-Tapasin

目的观察融合蛋白胞质转导肽(CTP)-HBcAg18-27-Tapasin诱导C57BL/6小鼠T淋巴细胞分泌Th1型细胞因子及HBV特异性细胞毒T淋巴细胞(CTL)的表达。方法 C57BL/6小鼠随机分为实验组CTP-HBcAg18-27-Tapasin、对照组CTP-HBcAg18-27、HBcAg18-27-Tapasin及空白组(生理盐水)。经肌肉免疫小鼠,ELISA检测T淋巴细胞分泌细胞因子;流式细胞术(FCM)检测T淋巴细胞内的细胞因子;CCK-8法检测T淋巴细胞增殖活性。结果实验组能有效刺激小鼠T细胞分泌Th1型细胞因子;FCM检测实验组融合蛋白诱导的CTL水平明显高于其他组;且实验组T淋巴细胞增殖活性明显高于对照组及空白组。结论 CTP-HBcAg18-27-Tapasin融合蛋白免疫C57BL/6小鼠后,能提高T淋巴细胞增殖活性,能有效刺激T淋巴细胞分泌Th1型细胞因子及增加CTLs的表达。

Objective To observe whether the fusion protein of cytoplasmic transduction peptide （CTP）-HBcAg18-27- Tapasin could induce Th1-type cytokine secretion and expression of hepatitis B virus （HBV） -specific cytotoxic T lymphocytes （CTL） in CS？BL/6 mice. Methods C57BL/6 mice were randomly divided into 4 groups, with 5 mice in each group. The mice of 4 groups were intramuscularly injected with the fusion protein CTP-HBcAg18-27-Tapasin, CTP-HBcAg18-27, HBcAg18-27-Tapasin （50 μg） and normal saline, respectively. The levels of cytokines secreted by T lymphocytes were detec- ted by ELISA and intracellular cytokine of proliferative T cells by flow cytometry. The proliferation of T lymphocytes was observed using CCK-8 assay. Results CTP-HBcAg^s_27-Tapasin not only induced significantly T lymphocyte proliferation, but also increased the production of cytokines （IFN-y, IL-2） compared with CTP-HBcAg18-27 or HBcAg18-27-Tapasin alone or nor- mal saline. Moreover, CTP-HBcAg18-27-Tapasin fusion protein increased significantly the percentages of IFN-y＋ CD8＋ T cells. Conclusion The CTP-HBcAg18-27-Tapasin fusion protein can effectively stimulate the secretion of Thl-type cytokines, induce T lymphocyte proliferation and increase the expression of HBV-specific CTLs in C57BL/6 mice.