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目的探讨不同免疫程序对表达乙型肝炎病毒(HBV)PreS2-S基因的DNA疫苗免疫效果的影响。方法分别采用重组质粒DNA疫苗单独免疫、重组MVA(r MVA)载体疫苗单独免疫、DNAprime/r MVAboost、DNAprime/r MVAboost并以表达鼠IL-18(mIL-18)的真核表达质粒为佐剂的程序免疫C57BL/6小鼠,通过ELISA检测抗-HB-sAb及ELISPOT检测IFN-γ评价体液免疫和细胞免疫的水平。结果表达PreS2-S抗原的各组疫苗在抗体产生的时间和滴度方面差异均无统计学意义,用ELISPOT检测IFN-γ的产生可知,所有免疫组均可引起针对特异性抗原的细胞免疫,r MVA单独免疫组强于DNA疫苗单独免疫组,弱于DNAprime/r MVAboost和DNAprime/r MVAboost+mIL-18组,DNAprime/r MVAboost和DNAprime/r MVAboost+mIL-18之间差异无统计学意义。结论携带HBV保护性抗原基因的重组MVA病毒可以诱导针对抗原的特异性细胞免疫和体液免疫,将其与表达相同抗原的DNA疫苗联合,采用DNAprime/rMVA boost的免疫方案,可以诱导比两者单独应用更强的细胞免疫反应。IL-18对免疫效果无明显影响。
Objective To investigate the effect of different immunization programs on the immune effect of DNA vaccine expressing hepatitis B virus PreS2-S gene. Methods Recombinant plasmid DNA vaccine was used to immunize mice with recombinant MVA (r MVA) vector alone. DNAprime / r MVAboost and DNAprime / r MVAboost were used as adjuvants in the eukaryotic expression plasmids expressing murine IL-18 (mIL-18) C57BL / 6 mice were immunized with anti-HB-sAb and ELISPOT by ELISA to assess the level of humoral and cellular immunity. Results There was no significant difference in antibody production time and titer among the groups of vaccine expressing PreS2-S antigen. Using ELISPOT to detect the production of IFN-γ, we could see that all immunized groups could induce cellular immunity against specific antigen, r MVA alone group was stronger than that of DNA vaccine alone and weaker than DNAprime / r MVAboost and DNAprime / r MVAboost + mIL-18 group. There was no significant difference between DNAprime / r MVAboost and DNAprime / r MVAboost + mIL-18 . Conclusion Recombinant MVA virus carrying HBV protective antigen gene can induce specific cellular immunity and humoral immunity against the antigen. Combined with the DNA vaccine expressing the same antigen, DNAprime / rMVA boost immunization program can induce the immune response than both Apply a stronger cellular immune response. IL-18 had no significant effect on the immune effect.