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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。
Objective To investigate the immune response of BALB / c mice induced by DNA vaccine containing Hantavirus L99 G2 glycoprotein gene. Methods The recombinant plasmid pVAX / G2 was transfected into cos-7 cells, and the transient expression products were detected by indirect immunofluorescence assay (IFA). The plasmid pVAX / G2 was extracted by alkaline lysis and purified by column chromatography on Sepharose CL-4B. BALB / c mice (SPF) were immunized with recombinant plasmid pVAX / IL-2 containing human IL-2 gene and recombinant human IL -2 protein as a DNA vaccine adjuvant to detect humoral and cellular immune responses in mice. Results The recombinant plasmid pVAX / G2 could be expressed in eukaryotic cells. Anti-Hantaan virus neutralizing antibody, enzyme-labeled antibody and fluorescent antibody can be detected in the serum of the immunized mice. The lymphocyte stimulation index and IL-2 activity in splenic lymphocyte supernatant were significantly different from those in empty vector pVAX1 negative control group. Plasmid adjuvant pVAX / IL-2 significantly increased the cellular immunity of DNA vaccines. Conclusion The DNA vaccine containing the G2 glycoprotein gene of Hantavirus L99 strain can induce some humoral and cellular immune responses in BALB / c mice.