HCV结构区DNA疫苗诱发小鼠特异性细胞免疫反应的研究(英文)

DNA immunization of mice with a plasmid encoding hepatitis C virus structural proteins elicits significant cell-mediated immune responses

目的通过 HCV结构区 DNA疫苗直接免疫小鼠 ,诱发其体内特异性细胞介导的免疫反应的研究 ,为 HCVDNA疫苗的研制打下基础。方法用重组的 p BK- CMV质粒体外转染小鼠骨髓瘤细胞 SP2 / 0 ,建立了能体外表达 HCV结构区蛋白的 SP2 / 0 D细胞系 ,分别用 SP2 / 0和 SP2 / 0 D细胞攻击用空质粒或 p BK- CMV质粒免疫过的 Balb/ c小鼠 ,通过肿瘤形成、肿瘤重量、肿瘤组织淋巴细胞的浸润以及不同免疫组小鼠血清中 IL - 2和 IFN -γ含量来观察小鼠体内 T淋巴细胞的活性。结果用 SP2 / 0和 SP2 / 0 D攻击空质粒或 p BK- CMV质粒免疫过的小鼠 15 d后 ,SP2 / 0 D攻击的 p BK- CMV质粒免疫鼠肿瘤重量 ( 0 .9± 0 .12 ) g明显低于空白质粒免疫组和 SP2 / 0接种组 ( P<0 .0 0 1) ,且该免疫组小鼠血清中 IL- 2和 IFN- γ的浓度明显高于其它免疫组 ;此外 ,用 SP2 / 0 D攻击的 p BK- CMV免疫鼠 ,肿瘤病理切片中可见明显的 T淋巴细胞浸润。结论重组的 HCV结构区 DNA疫苗 ( p BK- CMV)能诱导小鼠体内特异性 T淋巴细胞反应 ,HCV DNA疫苗可能为临床疾病的治疗和预防提供一种新的途径。

ObjectiveTo study the cell mediated immune responses in mice induced by direct plasmid pBK CMV immunization and to pave a way for further research of HCV DNA vaccine.MethodsWe used recombinant plasmid pBK CMV to transfect a syngeneic SP2/0 myeloma cells and established SP2/0D cell lines which can express HCV structural proteins. DNA immunized mice were inoculated with native SP2/0 or SP2/0D cells respectively. Activity of T lymphocytes was observed through tumor formation, tumor weight, T lymphocytes infiltration in tumor tissue and the level of IL 2,IFN γ in the serum of all immunized mice.ResultsInoculated with SP2/0D or native SP2/0 15 days later, tumor weight in the SP2/0D inoculated mice immnunized with pBK CMV plasmid was remarkably reduced compared with mice immunized with blank plasmid,or inoculated with SP2/0. The level of IL 2 and IFN γ in the serum of pBK CMV immunized groups were higher than those of blank plasmid immunized mice. In addition, the phenomena of T lymphocyte infiltration in tumor tissues were very apparent in the SP2/0D inoculated mice immnunized with pBK CMV plasmid.ConclusionsThe results indicate that recombinant plasmid pBK CMV can induce significant T lymphocytes activity in vivo by this technique, and HCV DNA immunization is a promising approach for vaccine development.