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

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

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，

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. \;