论文部分内容阅读
目的:观察重组表达质粒pcDNA3.1-S2S和pcDNA3.1-S1S2S对HBVDNA复制和抗原表达的抑制效果.方法:以能表达乙肝表面抗原主蛋白S(HBsAg)的重组真核表达质粒为骨架,构建了分别含有HBV前S1抗原和/或前S2抗原编码基因的pcDNA3.1-S1S2S,pcDNA3.1-S2S,并各以100μg肌肉接种C57BL/6HBVDNA转基因小鼠,2,4wk后各加强免疫1次.然后动态检测小鼠血清HBVDNA水平的变化、抗-HBs和前S2抗体的诱生,以及肝组织内HBsAg、前S2抗原的消长情况.结果:小鼠肝组织内HBsAg,前S2抗原呈逐渐减弱的趋势,8wk后各有1/3的小鼠已不能检出.pcDNA3.1-S1S2S组接种后4,8,12wk抗-HBs阳转率分别为50%、67%、100%,明显高于pcDNA3.1-S2S组(17%、33%、50%)(P<0.05);而前S2抗体阳转率均低于17%.接种后8wk,12wk,pcDNA3.1-S1S2S组和pcDNA3.1-S2S组小鼠血清HBVDNA水平明显下降,均低于自身接种前、接种后4wk以及同期对照组(P<0.05).结论:重组表达质粒pcDNA3.1-S2S和pcDNA3.1-S1S2S接种,能够抑制转基因小鼠体内的HBV复制,而重组质粒中S1基因的共表达使抑制作用得到增强.
OBJECTIVE: To observe the inhibitory effect of recombinant plasmid pcDNA3.1-S2S and pcDNA3.1-S1S2S on HBVDNA replication and antigen expression.Methods: The recombinant eukaryotic expression plasmid expressing HBsAg was used as the backbone, PcDNA3.1-S1S2S and pcDNA3.1-S2S containing the coding genes of pre-S1 antigen and / or pre-S2 antigen of HBV were constructed, and C57BL / 6HBVDNA transgenic mice were inoculated with 100μg of muscle respectively, Then the changes of serum HBV DNA levels, the induction of anti-HBs and pre-S2 antibodies, and the growth and decline of HBsAg and pre-S2 antigen in the liver tissue were detected dynamically.Results: The levels of HBsAg and pre-S2 antigen Gradually weakened trend, 1/3 of mice after 8wk can not be detected.PcDNA3.1-S1S2S group after vaccination 4, 8, 12wk positive rate of anti-HBs were 50%, 67%, 100% (17%, 33%, 50%) (P <0.05), while the positive rate of positive S2 antibody was less than 17% .After 8wk, 12wk, pcDNA3.1-S1S2S (P <0.05) .Conclusion: The expression of recombinant plasmid pcDNA3.1-S2S and pcDNA3.1-S2S were significantly lower than those of the control group (P <0.05) S1S2S Inoculation, can inhibit the HBV replication in transgenic mice, while the co-expression of the S1 gene in the recombinant plasmid makes the inhibition enhanced.