汉坦病毒DNA疫苗pVAX/G2诱导小鼠的免疫应答

目的探讨含汉坦病毒L99株G2糖蛋白基因的DNA疫苗诱导BALB/c小鼠的免疫应答。方法重组质粒pVAX/G2转染cos-7细胞,用间接免疫荧光法(IFA)检测瞬时表达产物。经碱裂解法提取质粒pVAX/G2,Sepharose CL-4B柱层析纯化后,免疫BALB/c小鼠(SPF),并采用含有人IL-2基因的重组质粒pVAX/IL-2及重组人IL-2蛋白作为DNA疫苗的佐剂,检测小鼠的体液免疫和细胞免疫应答。结果重组质粒pVAX/G2能够在真核细胞中表达。免疫小鼠血清中能够检测到抗汉坦病毒中和抗体、酶标抗体和荧光抗体。淋转刺激指数和脾淋巴细胞上清液中IL-2活性,与空载体pVAX1阴性对照比较差异有显著意义。质粒佐剂pVAX/IL-2能显著提高DNA疫苗的细胞免疫水平。结论含汉坦病毒L99株G2糖蛋白基因的DNA疫苗能诱导BALB/c小鼠产生一定强度的体液免疫和细胞免疫应答。

Immune Response Induced by Hantavirus DNA Vaccine in Mice

Objective To explore the immune response induced by DNA vaccine containing the gene encoding G2 protein of strain L99 of Hantavirus in BALB/c mice.Methods Transfect cos-7 cells with recombinant plasmid pVAX/G2 and detect the transiently expressed product by IFA.Extract plasmid pVAX/G2 by alkaline lysis and purify by Sepharose CL-4B column chromatography.Immunize BALB/c mice with the purified plasmid pVAX/G2 using recombinant plasmid pVAX/IL-2 and recombinant human IL-2 protein as adjuvant.Determine the humoral and cellular immune responses in the mice.Results Recombinant plasmid pVAX/G2 was expressed in eukaryotic cells.The neutralizing antibody,ELISA antibody and IFA antibody to Hantavirus were detected in the sera of immunized mice.The splenic lymphocyte transformation stimulating index(SI) and IL-2 activity in the supernatant of splenic lymphocyte of immunized mice were significantly different from that of negative control(empty vector pVAX1).As an adjuvant,plasmid pVAX/IL-2 enhanced the cellular immune response level significantly but failed to enhance the humoral immune response level.However,recombinant human IL-2 protein significantly enhanced both the humoral and cellular immune response levels.Conclusion The DNA vaccine containing the gene encoding G2 protein of strain L99 of Hantavirus induced both humoral and cellular immune responses in BALB/c mice.