中国寄生虫学与寄生虫病杂志 ›› 2004, Vol. 22 ›› Issue (3): 3-143.

• 论著 • 上一篇    下一篇

细胞因子表达质粒对恶性疟原虫顶端膜抗原1DNA免疫的调节作用

李珣1,缪军2,雷俊川2,薛采芳2,王宪锋2,刘忠湘2,李淑梅2
  

  1. 1 西藏军区总医院实验中心,拉萨 850003 2 第四军医大学病原生物学教研室,西安 710032
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2004-06-30 发布日期:2004-06-30

Modulation of Immune Response to Plasmodium falciparum Apical Membrane Antigen 1 DNA Vaccine by Cytokine Plasmids

LI Xun,MIAO Jun,LEI Jun-chuan,XUE Cai-fang,WANG Xian-feng,LIU Zhong-xiang,LI Shu-mei
  

  1. Research Center,General Hospital of Tibet Military Area,Lasa 850003,China
  • Received:1900-01-01 Revised:1900-01-01 Online:2004-06-30 Published:2004-06-30

摘要:   目的 探讨细胞因子表达质粒对小鼠DNA免疫的促进和调节作用。 方法 构建编码恶性疟原虫顶端膜抗原1(AMA1)完整胞外域的DNA免疫质粒VR1020/E,构建编码小鼠细胞因子(GMCSF)、白细胞介素(IL)如IL4和IL12的真核表达质粒pcDNA3/GMCSF、pcDNA3.1( ) /IL4和pIL12以及双顺反子质粒pGM CSF/pTPA E,分组免疫小鼠,ELISA检测血清中特异性IgG及其亚类的水平,取小鼠脾细胞进行体外增殖。 结果  3种细胞因子质粒均有效增强了小鼠针对VR10 20/E的免疫应答,抗体水平增加7至10倍,其中pcDNA3/GMCSF质粒和pIL12质粒分别显著促进了小鼠的IgG1和IgG2a应答,小鼠脾细胞的体外增殖水平亦有明显提高。 结论 利用编码GM CSF、IL4和IL12的表达质粒作为佐剂可有效增强小鼠针对AMA1DNA的免疫应答,并对免疫应答的类型产生调节作用。

关键词: 恶性疟原虫, 顶端膜抗原1, DNA疫苗, 细胞因子类, 佐剂, 免疫

Abstract:  Objective To explore the effect of cytokine encoding plasmids on DNA immunization in mice. Methods Prototype DNA vaccine VR1020/E which contain Plasmodium falciparum apical membrane antigen 1 (AMA1) ectodomain was constructed, and eukaryotic expression vectors pcDNA3/GM-CSF, pcDNA3.1(-)/IL-4, pIL-12 and pGM-CSF/pTPA-E were also built. BALB/c mice were immunized with VR1020/E alone or with VR1020/E plus different cytokine plasmids. Serum IgG and its subtype were determined by ELISA and in vitro splenocyte proliferation assay was done. Results GM-CSF, IL-4 and IL-12 encoding plasmids all promoted mice immune response to VR1020/E, the antibody level increased 7 to 10 times and splenocyte proliferation was enhanced too. Plasmid pcDNA3/GM-CSF induced much more IgG1 whereas plasmid pIL-12 induced much more IgG2a. Conclusion Cytokine encoding plasmids might be used as adjuvant in AMA1 DNA immunization.

Key words: Plasmodium falciparum, Apical membrane antigen 1, DNA vaccine, Cytokines, Adjuvant, Immunologic