西方马脑炎病毒E2蛋白的免疫原性研究

摘要

为研究西方马脑炎病毒(WEEV)囊膜E2蛋白的免疫原性,本研究通过诱导重组菌pET30-E2/E.coli BL21表达E2蛋白,将纯化的重组蛋白与弗氏佐剂乳化制备免疫原免疫小鼠,免疫剂量为50μg/只.二免后实验组小鼠体内CD4+/CD8+T细胞比例为3.38±0.19,细胞因子IL-2、IL-4及IFN-γ浓度分别为195.7±14.5 pg/mL、248.2±16.2 pg/mL与315.8±13.3 pg/mL;三免后实验组小鼠淋巴细胞增殖指数(PI)检测结果显示:ConA刺激组为2.18±0.17,E2蛋白刺激组为1.56±0.15;其特异性抗体IgG效价为1:640.以上实验组各项免疫指标均与对照组呈显著差异(p＜0.05),表明E2蛋白能够刺激小鼠产生较强免疫反应,具有较强免疫原性.本研究为WEEV基因工程亚单位疫苗研制奠定了良好基础.%In order to evaluate the immunogenicity of western equine encephalomyelitis virus (WEEV) E2 protein,the recombinant E2 protein was expressed in E.coli and purified according to His-bind protein purification kit.The BALB/c mouse was immunized with 50 μg E2 protein formulated with Freund's adjuvant.The detection results showed that the ratio of CD4+/CD8+T cell in immunized groups was 3.38±0.19,and the concentration of IL-2,IL-4 and IFN-γ was 195.7±14.5 pg/mL,248.2±16.2 pg/mL,315.8±13.3 pg/mL,respectively,post the secondary immunization.In addition,the proliferation index (PI) in immunized groups were 2.18±0.17 and 1.56±0.15 stimulated by ConA and E2 protein,respectively,and IgG antibody titers was 1:640 post the third immunization.Moreover,all the results above showed that the immune indices in immunized groups were significant difference compared with the control groups (p＜0.05),which demonstrated that E2 protein was able to induce efficiently immune response in immunized mice.