基础医学与临床 ›› 2018, Vol. 38 ›› Issue (5): 616-621.

• 研究论文 • 上一篇    下一篇

分泌型CTLA-4融合恶性疟原虫核酸疫苗联合GM-CSF增强小鼠免疫反应

高宇辉1,邓唯唯1,魏春燕2   

  1. 1. 中国医学科学院基础医学研究所 北京协和医学院基础学院
    2. 中国医学科学院基础医学研究所
  • 收稿日期:2018-01-26 修回日期:2018-03-20 出版日期:2018-05-05 发布日期:2018-04-28
  • 通讯作者: 高宇辉 E-mail:gaoyuhui@ibms.pumc.edu.cn
  • 基金资助:
    中国医学科学院医学与健康科技创新工程经费资助;国家自然科学基金

Secreted CTLA-4 fusion Plasmodium falciparum DNA vaccine combined with GM-CSF enhances immune response in mice

  • Received:2018-01-26 Revised:2018-03-20 Online:2018-05-05 Published:2018-04-28
  • Contact: Yh-Hui GAO E-mail:gaoyuhui@ibms.pumc.edu.cn

摘要: 目的 研究分泌型CTLA-4融合恶性疟原虫核酸疫苗联合GM-CSF免疫对小鼠体液免疫和细胞免疫反应的影响。方法 将疟原虫抗原编码序列与小鼠CTLA-4胞外区序列拼接构建真核表达载体VR1012-sES312-CTLA,用Western blot检测转染HEK293细胞上清液蛋白表达。VR1012-sES312-CTLA和小鼠GM-CSF的真核表达载体共同电穿孔法免疫Balb/c小鼠,ELISA检测疟疾疫苗特异性抗体IgG效价以及ELISPOT方法分析细胞因子IFN-γ和IL-4的表达水平。 结果 疟疾DNA疫苗体系中引入CTLA-4分子以及GM-CSF佐剂能显著增强疫苗的特异性免疫反应。VR1012-sES312-CTLA+GM-CSF联合免疫小鼠,抗体滴度比单纯的疟疾DNA疫苗VR1012-ES312相比提高190倍(P <0.001)。 结论 将疟疾DNA疫苗改造为树突细胞靶向型,并在免疫体系中引入GM-CSF分子佐剂,显著的增强了小鼠体液和细胞免疫效力。此结果为有效提高疟疾DNA疫苗免疫应答水平提供了新的思路。

关键词: 疟疾, DNA疫苗, 细胞毒性T淋巴细胞相关抗原-4, 粒细胞-巨噬细胞集落刺激因子

Abstract: Objective To study effects of secreted CTLA-4 fusion Plasmodium falciparum DNA vaccine combined with GM-CSF on the humoral and cellular immune responses in mice. Methods The malaria antigen coding sequence fused with CTLA-4 extracellular region of mouse to be constructed as eukaryotic secretory expression vector VR1012-sES312-CTLA, recombinant protein in culture of transfected HEK293 cells was detected by Western blot. Balb/c mice were co-administrated with VR1012-sES312-CTLA and GM-CSF expression vector. After immunization specific antibody IgG titers and cytokines IFN-γ and IL-4 expression levels were evaluated by ELISA and ELISPOT respectively. Results The introduction of CTLA-4 into malaria DNA vaccine system and application of GM-CSF adjuvant significantly enhanced the specific immune response of the vaccine. Antibody titers in VR1012-sES312-CTLA and GM-CSF co-immunized mice have a 190-fold increase compared with the simple designed VR1012-ES312 immunization (P <0.001). Conclusion humoral and cellular immunity of malaria DNA vaccine were both significantly enhanced by dendritic cell-targeting modification and the introduction of GM-CSF molecular adjuvant into the immune system. This result provides a new idea for effectively raising the immune response level of malaria DNA vaccine.

Key words: malaria, DNA vaccine, CTLA-4, GMCSF

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