PTD-HBcAg融合蛋白诱导特异性CTL在HepG2.2.15细胞抑制HBV复制的研究

doi:10.3969/j.issn.1672-5069.2010.03.002

实用肝脏病杂志 ›› 2010, Vol. 13 ›› Issue (3): 166-169.doi: 10.3969/j.issn.1672-5069.2010.03.002

PTD-HBcAg融合蛋白诱导特异性CTL在HepG2.2.15细胞抑制HBV复制的研究

赖静兰, 陈小华, 潘庆春, 臧国庆

200233 上海市上海交通大学附属第六人民医院感染病科

收稿日期:2010-01-25 出版日期:2010-06-10 发布日期:2016-04-18
通讯作者: 臧国庆,E-mail：zangguoqin@126.com
作者简介:赖静兰女,27岁,硕士研究生。主要从事慢性乙型肝炎的防治工作。E-mail：laijinglan.2931@yahoo.com.cn
基金资助:
国家自然科学基金资助项目（30571669）

Study on specific cytotoxic T lymphocytes induced by PTD-HBcAg fusion protein to inhibit HBV replication in HepG2215 cells

LAI Jinglan, CHEN Xiaohua, PAN Qingchun, et al.

Department of Infectious Disease,Sixth People,s Hospital,Jiaotong University,Shanghai 200233,China

Received:2010-01-25 Online:2010-06-10 Published:2016-04-18

摘要/Abstract

摘要： 目的探讨PTD-HBcAg融合蛋白诱导小鼠体内特异性CTL并抑制HepG2.2.15细胞HBV复制的作用。方法 PTD-HBcAg、HBcAg和阴性对照分别与等体积的弗氏佐剂乳化后皮下免疫小鼠;第14d,分离脾淋巴细胞并分别用PTD-HBcAg、HBcAg、PTD和PBS加强刺激后收集上清,检测细胞因子IFN-γ、IL-12、IL-4和IL-10; 刺激后的淋巴细胞作为效应细胞与HepG2.2.15细胞共培养,检测,效应细胞对HBsAg、HBV DNA的抑制作用及对HepG2.2.15细胞、HepG2细胞的杀伤效果。结果 PTD-HBcAg组分泌的IFN-γ、IL-12、IL-4和IL-10与HBcAg 组和阴性对照组比较差异有统计学意义（P<0.05）;PTD-HBcAg融合蛋白组较HBcAg组和阴性对照组有更明显的病毒抑制作用（P<0.05）;PTD-HBcAg组对HepG2.2.15细胞的杀伤率明显高于HBcAg组和阴性对照组（P<0.05）。结论 PTD-HBcAg可诱导HBV特异性CTL,能有效抑制HepG2.2.15细胞HBV的复制。

关键词: 乙型肝炎病毒, HepG2.2.15 细胞, PTD-HBcAg, 融合蛋白, 细胞毒性T淋巴细胞

Abstract: Objective To study the effects of PTD-HBcAg inducing HBV specific cytotoxic T lymphocytes in mice on inhibiting HBV replication in HepG2.2.15 cells. Methods BALB/c mice were subcutaneously immunized at the tail base with 100μl of freshly prepared emulsion containing with PTD-HBcAg,HBcAg,PTD,PBS and an equal volume of CFA. 14days later,T lymphocytes in spleen were isolated and stimulated with corresponding proteins to detect the level of IFN-γ,IL-12,IL-4 and IL-10 in the supernatants. The stimulated T lymphocytes as effective cells were cocultured with HepG2.2.15 cells to determine the levels of HBsAg and HBV DNA and to observe the cytotoxicity of effective cells to HepG2.2.15 cells. Results The cytokines in PTD-HBcAg groups were markedly higher than HBcAg groups and normal controlled groups;In comparison with normal controlled groups and HBcAg groups,HBsAg and HBV DNA decreased more obviously in PTD-HBcAg groups;The T lymphocytes stimulated by PTD-HBcAg could kill HepG2.2.15 cells more effectively than others. Conclusions PTD-HBcAg can highly promote HBV specific CTL to inhibit HBV replication in HepG2.2.15 cells.

Key words: Hepatitis B virus, PTD-HBcAg, Fusion protein, Cytotoxic T lymphocyte, HepG2.2.15 cell line

赖静兰, 陈小华, 潘庆春, 臧国庆. PTD-HBcAg融合蛋白诱导特异性CTL在HepG2.2.15细胞抑制HBV复制的研究[J]. 实用肝脏病杂志, 2010, 13(3): 166-169.

LAI Jinglan, CHEN Xiaohua, PAN Qingchun, et al.. Study on specific cytotoxic T lymphocytes induced by PTD-HBcAg fusion protein to inhibit HBV replication in HepG2215 cells[J]. JOURNAL OF PRACTICAL HEPATOLOGY, 2010, 13(3): 166-169.

参考文献

[1] REHERMANN B. Intrahepatic T cells in hepatitis B：viral control versus liver cell injury[J]. Exp Med,2000,191：1263-1268.
[2] STEWART KM,HORTON KL,KELLEY SO.Cell-penetrating peptides as delivery vehicles for biology and medicine[J]. Org Biomol Chem,2008,6（13）：2242-2255.
[3] 潘庆春,臧国庆,余永胜,等. 转录反式激活蛋白转导域介导HBcAg穿透细胞膜的作用[J]. 中华传染病杂志,2008,26：336-340.
[4] DELOM F,FESSART D,CARUSO ME,et a1. Tat-mediated protein delivery in living Caenorhabditis elegans[J]. Biochem Biophys Res Commun,2007,352（3）：587-591.
[5] CHEN XH,YU YS,PAN QC,et al.Enhancement of cytotoxic T lymphocyte activity by dendritic cells loaded with Tat-PTD fused HbcAg[J]. Acta Biochimica Biophysica Sinica,2008,40（12）：996-1004.
[6] ALBARRAN B,TO R,STAYTON PS.A TAT-streptavidin fusion protein directs uptake of biotinylated cargo into mammalian cells[J]. Protein Eng Des Sel,2005,18（3）：147-152.
[7] 陈小华,潘庆春,余永胜,等. PTD-HBcAg体外诱导小鼠髓源性树突状细胞成熟及在T淋巴细胞增值中的作用[J]. 中华传染病杂志,2009,27（4）：198-202.
[8] YE ZM,TANG C,XU CL. Type 1 CD8+T cells are superior to type2 CD8+T cells in tumor immunotherapy due to their efficient cytotoxicity,prolonged survival and type1 immune modulation[J]. Cell Mol Immunol,2007,4（4）：277-285.
[9] JUSTESEN S,BUUS S,CLAESSON MH,et a1. Addition of TAT protein transduction domain and GrpE to haman p53 provides soluble fusion proteins that can be transduced into dendritic cells and elicit p53-specific T-cell responses in HLA-A*0201 transgenic mice[J]. Inmmnology,2007,122（3）：326-334.
[10] VIEHL CT,TANAKA Y,CHEN TT,et al. Tat mammaglobin fusion protein transduced dendritic cells stimulate mammaglobin-specific CD4 and CD8 T cells[J]. Breast Cancer Res Treat,2005,91：271-278.

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PTD-HBcAg融合蛋白诱导特异性CTL在HepG2.2.15细胞抑制HBV复制的研究

Study on specific cytotoxic T lymphocytes induced by PTD-HBcAg fusion protein to inhibit HBV replication in HepG2215 cells

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