丙型肝炎病毒DNA核酸免疫的初步探讨

窦　骏　刘克洲　陈　智　沃建尔　刘　勇　徐陈槐　何南祥　章明太　王信子

　　中国图书分类号　R512.63
　　摘　要　目的：探索HCV-DNA核酸免疫效应。方法：将构建的重组体pCD-HCVs免疫Balb/c小鼠，用ELISA和3H-TdR掺入法检测了免疫鼠血清抗体、脾细胞和PBMC对HCV抗原的增殖反应。结果：重组体pCD-HCV1、pCD-HCV2和pCD-HCV3均可诱导小鼠(n=6)产生抗体(0.665～0.707，OD值)和脾细胞对HCV抗原增殖反应(n=6,SI为3.89～4.47)，但各重组体间的免疫效应无显著性差异(P＞0.05);免疫鼠(n=12)血清经40倍和320倍稀释后，抗体阳性率分别为100.0%和16.6%；重组体还可诱导鼠PBMC对HCV抗原的增殖反应(3.69～4.07，SI)。结论：pCD-HCVs重组体可诱导小鼠产生免疫反应，该结果对进一步开展HCV-DNA疫苗研制将具有重要意义。
　　关键词　HCV-DNA重组体　核酸免疫　抗体反应　细胞免疫

Preliminary inquire into approach of HCV-DNA nucleicacid immunity
DOU Jun,LIU Ke-Zhou,CHEN Zhi et al.Institute of Infectious Disease,Zhejiang Medical University,Hangzhou 310003

　　Abstract Objective:To explore the effect of HCV-DNA nucleicacid immunity.Methods:Recombinant pCD-HCVs constructs were injected into muscles of Balb/c mice and serum anti-HCV were tested by ELISA(OD value).Spleen cells and PBMC proliferation responses to HCV antigens were detected by 3　H-TdR incorporation method(cpm).Results:Recombinant pCD-HCV1,pCD-HCV2and pCD-HCV3 can all elicite mice to generate antibody(0.665～0.707，OD value) and spleen cells in mice (n=6) were induced to be proliferative responses to HCV antigens(3.89～4.47)，stimulation index(SI).There were no significant difference(P＞0.05)among each recombinant,the serum antibody in mice(n=12) immunized by recombinant pCDHCVs were 100.0 percent positive and 16.6 percent positive when serum were diludted 40 flods and 320 flods respectively.PBMC proliferative responses to HCV antigens in mice immunized with pCD-HCVs was detected also(3.69～4.07,SI).Conclusion:The investigations suggested that recombinant pCD-HCVs can elicite Balb/c mice to generate immune response and the experiment results have an important significant for development of HCV-DNA vaccine in future.
　　Key words HCV-DNA recombinant Nucleicacid immunity Antibody responses Cellular immunity

　　　自1993年Ulmer等首次报道DNA免疫技术用于流感病毒疫苗研制以来［1］，肝炎病毒等DNA疫苗的研究已逐步展开［2］。为探索我国丙型肝炎病毒（H

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