中国寄生虫学与寄生虫病杂志 ›› 1999, Vol. 17 ›› Issue (6): 2-329.

• 论著 • 上一篇    下一篇

恶性疟原虫FCC1/HN株CSP抗原基因表达产物在小鼠体内的免疫应答研究

方政;余新炳;刘彦文;罗树红
  

  1. 中山医科大学寄生虫学研究所 广州 510089
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:1999-12-30 发布日期:1999-12-30

STUDIES ON IMMUNE RESPONSES IN MICE TO EXPRESSED PRODUCTS FROM CSP GENE OF PLASMODIUM FALCIPARUM FCC1/HN ISOLATE\+*

FANG Zheng **;YU Xinbing;LIU Yanwen;LUO Shuhong


  

  1. Institute of Parasitology;Sun Yat-Sen University of Medical Sciences;Guangzhou 510089
  • Received:1900-01-01 Revised:1900-01-01 Online:1999-12-30 Published:1999-12-30

摘要:   目的: 利用pcDNA3 质粒作为载体, 在人宫颈癌细胞(HeLa 细胞) 中高效表达恶性疟原虫环子孢子蛋白 (CSP), 观察表达产物诱导BALB/c小鼠免疫的应答水平。方法: 目的基因PfCSP在HeLa 细胞中表达, 将纯化的表达产物免疫接种小鼠, 通过ELISA、Western blotting 分析、T 淋巴细胞增殖实验、NK 细胞活性检测和T淋巴细胞亚群测定, 观察其诱导BALB/c小鼠产生体液免疫和细胞免疫的应答水平。结果: ELISA 检测抗体滴度达1∶6 400;Western blotting 结果在38.3 kDa 相应位置出现较清晰的显色条带;表达产物能特异刺激小鼠脾淋巴细胞增殖、CD4+ 与CD8+ 细胞有所增加, 并能提高小鼠NK细胞活性。结论: 真核表达系统pcDNA3-Pf/HeLa表达产物能特异刺激小鼠产生体液免疫和细胞免疫的应答, 并提高小鼠NK细胞活性的作用

关键词: 恶性疟原虫, 环子孢子蛋白, HeLa细胞, 表达产物, 免疫应答

Abstract:  AIM: To construct the eukaryotic expression system with pcDNA3-PfCSP/HeLa for CSP gene of \{Plasmodium\} falciparum, and to observe the immune responses in BALB/c mice induced by the expressed proteins. \{METHODS\}: The recombinant plasmid pcDNA3-PfCSP was transformed into mammalian cell line of human HeLa cells. The expressed protein was isolated and analyzed by SDS-PAGE and used for immunization of BALB/c mice by subcutaneous, intravenous or intraperitoneal administration, respectively. ELISA, Western blotting, T lymphocyte proliferation test, natural killer cell activity and CD4\++ and CD8\++ T cells detection were used for observation of the level of humoral and cellular immune responses. RESULTS: Immune sera strongly reacted with the expressed protein. The titer of the antibodies was up to 1∶6 400 by ELISA. Western blotting analysis revealed a specific band at 38.3 kDa. The specific proliferative response with immunized BALB/c mice spleen cells was remarkablely higher than that with control ones. The ratio of CD4\++ and CD8\++ T cells and the NK cell activity were significantly higher in the immunization group than that in the control groups. CONCLUSION: The humoral and cell-mediated immune responses and elevated NKC activity to expressed products with the eukaryotic expression system can specifically be detected in BALB/c mice to generate, indicating that the expressed protein could enhance immune activity in mice. \;

Key words: Plasmodium falciparum, circumsporozoite protein, HeLa cell, expressed product, immune response