重组乙型肝炎病毒变异s基因疫苗诱导小鼠产生特异性体液免疫应答

Induction of specific humoral immune response in mice by recombinant mutant HBV s gene vaccine

目的 :构建乙型肝炎病毒 (HBV)变异s基因真核表达载体 ,检测其诱导小鼠产生特异性体液免疫应答。方法 :利用限制性内切酶定位克隆构建s基因nt5 87G→A的真核表达载体pCMV S2 .S +14 5R(PR)。用其转染人肝癌细胞系HepG2后 ,用EIA、ELISA及免疫细胞化学法 ,观察其抗原性。以重组变异型s基因真核表达载体 (PR)和载体pcDNA3.0分别免疫C5 7BL/6小鼠各 5只。每只小鼠各肌肉注射纯化质粒 10 0 μg。用ELISA法检测血清抗 HBs及抗 HBs2抗体的效价。结果 :体外实验证实 ,变异型HBsAg可与抗 HBs结合 ;PR免疫小鼠可诱导其产生抗 HBs抗体及抗 HBs2抗体 ,但抗 HBs2抗体的出现早于抗 HBs抗体约 1～ 2wk。结论 :HBV变异s基因(nt5 87G→A)的真核表达载体的表达产物具有良好的抗原性 ,能够诱导C5

AIM: To construct the recombinant eukaryotic expression vector pCMV S2.S+145R(PR) containing mutant HBV s gene and detect the specific humoral immune response to the PR in mice. METHODS: The PR was constructed by positional cloning of restriction endonuclease, and then it was transfected into human hepatocellular carcinoma cell line Hep G2 through electrotransformation. The antigenicity of PR was examined by EIA, ELISA and immunocytochemical staining. The PR and empty vector pcDNA3.0 were then used respectively to immunize intramuscularly 5 C57BL/6 mice, dosage being 100 μg purified plasmid each mouse. The titers of serum anti HBs and anti HBs2 antibodies were detected by ELISA. RESULTS: In vitro experiment showed that mutant HBsAg could bind to anti HBs antibody. The PR could induce anti HBs and anti HBs2 antibody production in immunized mice. But the appearance time of serum anti HBs2 antibody one or two weeks eariler than that of serum anti HBs antibody. CONCLUSION: The expression product of PR had a good antigencity, which can induce specific humoral immune response in C57BL/6 mice.