Yin Xiao, Wang Wen, Tan Wen-jie, Deng Yao, Guan Jie, Wen Bo, Chen Hong, Ruan Li
Biotech Center for Viral Diseases Emergency, National Institute for Viral Disease Control and Prevention, China CDC, Beijing 100052, China.
Bing Du Xue Bao. 2011 Jan;27(1):44-9.
To develop a new Hepatitis C virus (HCV) DNA vaccine ,We explored strategies for optimizing the immunogenicity of HCV DNA vaccine in fusion with dendritic cell-targeting molecules (scDEC, a single-chain antibody against the murine DC cell surface molecule DEC205). We constructed the DNA vaccine plasmids expressing the HCV non-structural protein NS3 alone or in combination with DEC205 as a fusion protein and identified the expression of the molecules of interest by transient transfection of 293 cells with the resultant DNA vaccine plasmids. Then BALB/C mice were immunized with these plasmids by the injection in combination with electroporation. The NS3-specific IgG antibody(ELISA) and cellular immunity (IFN-gamma ELISPOT) were analyzed post twice immunization. Our results showed that: the single-chain antibody against DEC205 fused with vaccine antigen NS3 significantly enhanced the immunogenicity of new HCV DNA vaccine, the intradermal injection in combination with electroporation using caliper electrodes resulted in most robust NS3-specific antibody and T cell immune response. In conclusion,immune response for the HCV NS3 protein-encoding DNA vaccine was enhanced significantly when targeting antigen NS3 to DCs by scDEC. The present strategy could merit further study in the context of other prophylactic and therapeutic DNA based vaccines.
为研发一种新型丙型肝炎病毒(HCV)DNA疫苗,我们探索了优化HCV DNA疫苗与树突状细胞靶向分子(scDEC,一种针对小鼠DC细胞表面分子DEC205的单链抗体)融合后的免疫原性的策略。我们构建了单独表达HCV非结构蛋白NS3或与DEC205作为融合蛋白共同表达的DNA疫苗质粒,并通过用所得DNA疫苗质粒瞬时转染293细胞来鉴定目的分子的表达。然后,通过联合电穿孔注射用这些质粒免疫BALB/C小鼠。在两次免疫后分析NS3特异性IgG抗体(ELISA)和细胞免疫(IFN-γ ELISPOT)。我们的结果表明:与疫苗抗原NS3融合的针对DEC205的单链抗体显著增强了新型HCV DNA疫苗的免疫原性,使用卡尺电极联合电穿孔进行皮内注射产生了最强的NS3特异性抗体和T细胞免疫反应。总之,当通过scDEC将抗原NS3靶向DC时,编码HCV NS3蛋白的DNA疫苗的免疫反应显著增强。本策略在其他基于DNA的预防性和治疗性疫苗的背景下值得进一步研究。