摘要目的 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HBV DNA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.

abstractsObjective 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.