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目的评价融合蛋白基因质粒(pFP)联合在体电脉冲(EP)介导的HBVDNA疫苗(pS2.S)免疫HBV转基因(Tg)小鼠的治疗效果。方法HBVTg小鼠随机分成3组,每组5只,分别为pS2.S+pFP、pS2.S+pcDNA3.1免疫治疗和pcDNA3.1对照组,联合免疫质粒总剂量40μg/只,按1∶1比例混合接种。初次免疫后第4、8周分别进行第一、二次增强免疫,免疫前后分别检测血清学、组织学及HBV特异性免疫应答。结果HBVDNA血清定量检测结果,pS2.S+pFP组免疫第8周(13317±2539)拷贝/ml、第12周(6462±3359)拷贝/ml时分别较免疫前(36159±7769)拷贝/ml明显减低,差异具有统计学意义(P<0.01);pS2.S+pcDNA3.1组第4周(20618±9523)拷贝/ml及第8周(23818±5319)拷贝/ml时均较其免疫前水平(36090±4421)拷贝/ml明显降低(P<0.01),但不能持续到12周(27691±13071)拷贝/ml,并明显高于此时pS2.S+pFP组水平,差异有统计学意义(P<0.01);观察终点(12周)pS2.S+pFP组Tg小鼠个体血清HBVDNA及肝组织HBsAg表达水平降低的同时,伴随其血清ALT水平及HBsAg特异性IFN-γ分泌细胞数目的升高。结论Th1类细胞因子IL-2/IFN-γ融合蛋白基因表达质粒能够增强HBVDNA疫苗抑制Tg鼠HBVDNA复制和表达的治疗作用。
Objective To evaluate the therapeutic effect of the fusion protein gene plasmid (pFP) in combination with HBV vaccine (pS2.S) immunization of HBV transgenic mice induced by EP (EP). Methods HBVTg mice were randomly divided into 3 groups with 5 mice in each group, which were pS2.S + pFP, pS2.S + pcDNA3.1 immunotherapy and pcDNA3.1 control group. The total combined immunophenotype dose was 40μg / 1 ratio mixed inoculation. After the first immunization, the first and second booster immunizations were performed on the 4th and 8th week respectively. The serological, histological and HBV specific immune responses were detected before and after immunization. Results Compared with the pre-immunization (36159 ± 7769) copies / ml at the 12th week (6462 ± 3359) copies / ml in the pS2.S + pFP group at the eighth week (13317 ± 2539) copies / (P <0.01). Compared with that of pS2.S + pcDNA3.1 group at the fourth week (20618 ± 9523) copies / ml and the eighth week (23818 ± 5319) copies / ml The former level (36090 ± 4421) copies / ml was significantly lower (P <0.01), but could not last for 12 weeks (27691 ± 13071) copies / ml and was significantly higher than that of pS2.S + pFP group at this time (P <0.01). At the end of the study (12 weeks), the levels of serum HBVDNA and HBsAg in Tg mice with pS2.S + pFP decreased as well as the levels of serum ALT and HBsAg-specific IFN-γ secreting cells The increase in the number. Conclusion Th1 cytokine IL-2 / IFN-γ fusion protein gene expression plasmid can enhance the therapeutic effect of HBVDNA vaccine on the replication and expression of HBVDNA in Tg rats.