乙型肝炎病毒DNA疫苗电转加强含S-PreS1颗粒的蛋白疫苗在小鼠中免疫效果的研究

目的探索新型有效的HBV治疗性疫苗研制的策略。方法构建含S与PreS1融合基因的HBVDNA疫苗，采用两次蛋白疫苗（含不同佐剂）肌内注射初免，一次DNA疫苗皮内电转免疫加强的联合免疫方式，在小鼠中分析比较了各疫苗所引起的体液与细胞免疫应答。结果HBVDNA疫苗皮内注射加电转可明显增强表面抗原（s）特异的细胞免疫应答（IFN-γ ELISpot分析）及PreS1特异性抗体水平，并以蛋白疫苗加铝佐剂初免组细胞免疫增强效果最明显。结论HBSS1 DNA疫苗结合皮内注射＋电转方式可明显加强含S-PreS1颗粒的蛋白疫苗在小鼠中免疫效果，明显高于蛋白疫苗单独应用。这些研究结果为新型HBV治疗性疫苗的研制与应用提供了依据。

HBV DNA vaccination with electroporation enhances significantly the specific cell-mediated immune response in mice against HBV protein vaccine consisting of S-PreS1 fusion particles

Objective To rational design HBV therapeutic vaccine candidate and evaluate their specific immunity to HBV in mice.Methods Based on our previous data of HBV protein vaccine consisting of S-PreS1 fusion particle.We first construct a novel DNA vaccine candidate,pVRC-HBSS1,which consisting of S (an:1-223) and PreS1 (an:21-47) fuse gene,then confirm the expression of the DNA vaccine by Western blotting,and followed by vaccination using prime boost strategy,ie,Intradermal injection of DNA vaccine with gene electroporation (EP) in BALB/c mice after twice injection of different HBSS1 protein vaccines (combination with different adjuvants).The immune response was measured by ELISA and IFN-gamma ELISPOT.Result The novel DNA vaccine candidate could effectively express in vitro,boost with single intradermal injection of HBV DNA vaccine via EP can significantly enhance the surface antigen (S)-specific cellular immune responses (IFN-γ,ELISpot analysis) and PreS1-specific antibody levels,especially in the group primed with protein vaccine in combination with alum adjuvant.Conclusion Boost with the novel HBV DNA vaccine followed prime with HBV protein vaccine could induced a higher anti-HBV T cell response in mice than vaccination with the HBSS1 particle-like protein vaccine only.This prime-boost vaccination may serve as a promising way to develop and optimize the novel HBV therapeutic vaccine.