乙肝病毒前S/S抗原的重组表达质粒对HBV转基因小鼠的免疫治疗效应  

Immune therapeutical effect induced by inoculation with recombinant plasmid encoding preS/S antigen in HBV DNA transgenic mice

作　　者：周陶友[1] 陈敏[1] 赵连三[1] 王松[2] 陈守春 何芳[1] 刘丽[1] 唐红[1] 

机构地区：[1]四川大学华西医院感染性疾病中心人类疾病生物治疗国家重点实验室,四川省成都市610041 [2]深圳东湖医院内科,广东省深圳市518000 [3]成都地奥集团药物研究所,四川省成都市610041

出　　处：《世界华人消化杂志》2005年第7期844-847,共4页World Chinese Journal of Digestology

基　　金：国家自然科学基金项目;No.39070670~~

摘　　要：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.AIM: To evaluate the effects of recombinant plasmids pcDNA3.1-S1S2S and pcDNA3.1-S2S encoding hepatitis B virus (HBV) envelope S/preS1 or S/preS2 antigens on the inhibition of HBV DNA replication and antigen expression in HBV transgenic mice. METHODS: Plasmids pcDNA3.1-S1S2S and pcDNA3.1-S2S were constructed. 100 μg of each plasmid was injected muscularly to inoculate C57BL/6 HBV DNA transgenic mice. The inoculation was boosted 2 and 4 weeks later. Serum samples were collected to measure anti-HBs, anti-HBc and HBV DNA. The liver tissue biopsies were performed to detect HBsAg and preS2 antigen. RESULTS: Four, 8 and 12 weeks after inoculation, the positive rates of anti-HBs in pcDNA3.1-S1S2S group were 50%, 67% and 100%, respectively, which were higher than those in pcDNA3.1-S2S group (16.7%, 33%, 67%) (P<0.05). The positive rates of anti-preS2 were lower than 16.7% in both groups. The numbers of HbsAg or preS2 antigen positive hepatocytes were also decreased significantly in both groups. The serum HBV DNA titers at 8,12 weeks were lower than those before and 4 weeks after the first inoculation. No change was observed in the control mice, which were inoculated with the empty plasmid vector. CONCLUSION: The data showed that pcDNA3.1-S1S2S and pcDNA3.1-S2S inoculation might inhibil replicalion of HBVDNA and expression of HbsAg and preS2 antigen in mice. The preS1 insert may enhance the effect of pcDNA3.1 -S2S. It is suggested that pcDNA3.1-S1S2S and pcDNA3.1-S2S can be DNA vaccine candidates for the treatment of chronic HBV infection.

关 键 词：重组表达质粒 HBV转基因小鼠 治疗效应 乙肝病毒 pcDNA3 S抗原 HBV前S1抗原 抗-HBS阳转率 前S2抗原 表面抗原主蛋白 C57BL/6 DNA水平 血清HBV HBsAg 真核表达质粒 DNA复制 前S2抗体 抗体阳转率 HBV复制 接种后 抑制效果 

分 类 号：R392[医药卫生—免疫学]