丙型肝炎病毒DNA核酸免疫的初步探讨

Preliminary inquire into approach of HCV DNA nucleicacid immunity

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。

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 HCV 1,pCD HCV 2and pCD HCV 3 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.