HBV患者树突状细胞(DC)数量减少和功能缺陷，不能产生有效的针对HBV特异的CTL反应，导致病毒持续复制不能被清除，是HBV长期感染的重要原因。MIP-3α 是DC最重要的特异性趋化因子，能够有效提高DC 递呈抗原的能力。HBx是抗HBV治疗的重要靶点，针对 HBV X 区设计siRNA 可能是清除病毒的最佳选择。本项目利用重组PCR技术构建靶向HBx的siRNA；利用反向遗传（病毒拯救）技术构建高效表达MIP-3α的重组病毒AAV/MIP-3α，借助AAV-Helper-free表达系统构建AAV-MIP3α-HBx- siRNA重组病毒质粒。转染DC细胞诱导产生高效特异抗HBV的CTL细胞。分别通过人肝癌细胞HepG2细胞、转染HBV质粒的HepG2.2.15细胞和M-TgHBV转基因小鼠乙肝模型，探讨ACTL抗HBV的治疗效果及免疫学机制，为抗HBV的临床治疗提供新的策略和思路。

The reason of the Long-term infection with HBV is the decreased in the number of dendritic cells (DC) or functional defects, the cell can not produce the special cytotoxic lymphocyte response to against HBV. Thus, HBV can persistent replicate inside the liver cells in the long-term infection. MIP-3α is the most important chemotactic factor, which can enhance antigen uptake and antigen processing of DC. The HBV X (HBx) gene is consistently expressed in all four HBV viral mRNAs and thus an ideal target for developing viral inhibitors via a gene therapeutic approach. small interfering RNAs(siRNA) could decrease both viral RNA and protein levels, and completely block replication in cultured cells co-transfected with a siRNA expression plasmid and an HBV replication-competent vector. In the project, Generation and functional analyze HBV specific CTL elicited by DC transfected with recombinant adenovirus in vitro. Using recombination DNA technique construct HBx-targeted siRNA, the fusion expression plasmid of AAV/MIP-3α, and siRNA expression cassettes (SECs) containing AAV-MIP3α-HBx- siRNA. The human hepatoma cell HepG2 and M-TgHBV transgenic mice for evaluation of drugs treating hepatitis B virus is used to investigate the ACTL treatment anti-HBV effect and immunological mechanism, It also will lead to new strategies for therapies.