丙型肝炎病毒多CTL表位DNA疫苗免疫学特性的初步研究

目的:探讨丙型肝炎病毒(HCV)疫苗研制并为抗HCV感染提供实验依据。方法:从HCV J株基因组中,选取小鼠(H-2d)CTL表位基因序列,以pcDNA3.1(+)质粒为载体,构建相应的真核表达质粒,经肌肉免疫后,取受免疫小鼠血清和脾细胞,用ELISA及MTT法检测小鼠血清中IL-2和IFN水平及小鼠脾细胞的增殖活性。结果:实验组小鼠血清中IL-2及IFN水平明显高于对照组IL-2:(208±8.88)比(106±6.06);IFN:(179.5±2.52)比(90.5±2.52),P<0.05];实验组淋巴细胞增殖指数明显高于对照组(1.56比1.13,P<0.05)。结论:本实验构建的pcDNA3.1(+)/HCV CTL真核表达质粒,经肌肉免疫后可以使受免疫小鼠细胞免疫应答水平提高,淋巴细胞增殖活性增高。

Investigating Potential Immunogenic Efficacy of HCV DNA Vaccine with Multiple CTL Epitopes

Objective: To explore the experiment evidence for further research on hepatitis C virus(HCV) vaccine and anti-HCV infection.Methods: The candidate HCV DNA vaccine was constructed by inserting the H-2d mice CTL epitopes sequences from HCV J strain genome into eukaryotic expression plasmid pcDNA3.1(+),and BalB/c mice were immunitied intra-muscle with the pcDNA3.1(+)plasmid.The mice sera were collected for the test of IL-2 and INF levels by ELISA,and spleen cells were isolated for lymphocyte proliferation by MTT method.(Results:) Serum IL-2 and IFN levels in experiment group were higher than those in control group(IL-2:208±8.88 versus 106±6.06;IFN:179.5±2.52 versus 90.5±2.52;both P<0.05);Lymphocyte proliferation index was higher in experiment group than that in control group((P<0.05).) (Conclusion:) The pcDNA3.1(+)/HCV CTL eukaryotic expression plasmid was constructed successfully in this experiment.By inoculation,the recombinant plasmid pcDNA3.1(+)/HCV CTL can enhance cell-mediated immune response in mice,and induce lymphocyte proliferation response to specific antigen.