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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。
Objective To directly immunize mice with DNA vaccine of HCV region to induce specific cell-mediated immune responses in vivo, and to lay the foundation for the development of HCVDNA vaccine. Methods SP2 / 0 mouse myeloma cells were transfected with recombinant plasmid pBK-CMV in vitro. SP2 / 0 D cell line expressing HCV structural protein in vitro was established and challenged with SP2 / 0 and SP2 / 0 D cells respectively Balb / c mice immunized with empty plasmid or pBK-CMV plasmid were observed by tumor formation, tumor weight, infiltration of tumor tissue lymphocytes, and IL-2 and IFN-γ levels in sera from different immunized groups Rat T lymphocyte activity. Results The mice immunized with SP2 / 0 and SP2 / 0 D challenged with either pBK-CMV plasmid or pBK-CMV plasmid for 15 days, and the SP2 / 0D-challenged pBK-CMV plasmid immunized mice with tumor weight (0.9 ± 0. 12) g was significantly lower than that of the blank and SP2 / 0 groups (P <0.01). The concentrations of IL-2 and IFN-γ in serum of the immunized mice were significantly higher than those of the other immunized groups. In addition, pBK-CMV mice challenged with SP2 / 0 D showed significant T lymphocyte infiltration in the tumor pathological sections. Conclusion Recombinant HCV DNA vaccine (pBK-CMV) can induce specific T lymphocyte response in mice. HCV DNA vaccine may provide a new approach for the treatment and prevention of clinical diseases.