中国流行株HIV-1Gag与IFN-α2b重组质粒的构建与实验免疫研究

目的探讨中国流行株 ＨＩＶ－ １Ｇａｇ与 ＩＦＮ－ α２ｂ共表达重组核酸疫苗质粒的免疫效果。方法以 核酸疫苗质粒ｐＩＲＥＳ１ｎｅｏ为表达载体，构建重组核酸疫苗质粒ｐＩＲＥＳ１－Ｇａｇ、ｐＩＲＥＳ１－Ｇａｇ－ＩＦＮ，通过间接免疫荧光 试验、ＤｏｔＥＬＩＳＡ检测Ｃａｇ／ｈＩＬ－２／ｈＩＬ－６基因的表达产物。另将此重组核酸疫苗质粒免疫ＢＡＬＢ／ｃ小鼠，进行淋巴细 胞转化试验、ＣＤ４＋、ＣＤ８＋Ｔ淋巴细胞数量测定、细胞毒性Ｔ淋巴细胞（ＣＴＬ）特异性杀伤作用检测及血清抗体检测。 结果构建的重组质粒转染ＢＨＫ细胞后可表达目的基因，免疫小鼠后可有效地刺激淋巴细胞增殖，诱导特异性 ＣＴＬ反应，当和 ＩＦＮ－α２ｂ共表达时免疫效果更加显著。结论与 Ｇａｇ蛋白共表达的 ＩＦＮ－ａ２ｂ能够进一步提高细胞 免疫与体液免疫水平，构建的重组质粒为ＨＩＶ－１ＤＮＡ疫苗的研究奠定了一定实验基础。

Construction and Immunological Study of Recombinant Plasmid with HIV-1Gag and IFN-

Objective To research the immunity of recombinant HIV-1 DNA vaccine plasmids in mice.Methods Two DNA vaccine recombinant plasmids,pIRES1-Gag and pIRES1-Gag-hIL-IFN,were constructed by inserting the HIV-1 Gag and IFN-a2b genes into a DNA vaccine expression vector pIRES1 neo.The expressed product o gag/IFN-a2b gene was examined by Dot-ELISA and indirect immunofluorescence.Immune responses of BALB/c mice immunized with the 2 DNA vaccine recombinant plasmids were examined by lymphocyte transformation test(LTT),cytotoxic T lymphocyte specific killer test,quantitative analysis of Tlymphocyte (CD4 ,CD8 )and dynamics of specific serum IgG antibody.Results High expression was observed in BHK cells transfected with pIRES1-Gag and pIRES1-Gag -IFN.The co-expressed product of Gag and IFN-a2b genes showed more significant effects.Conclusion Cytokine IFN-a2b can enhance both the humoral and cellular immunity in mice.The recombinant plasmid is suitable for developing HIV-1 DNA vaccine.