| SAG1-MIC8复合DNA基因疫苗免疫小鼠诱导抗弓形虫感染的保护性研究(英文) |
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| 引用本文： | 姚远,何深一,王花欣,周怀瑜,赵红,李婷,薛明福,朱兴全. SAG1-MIC8复合DNA基因疫苗免疫小鼠诱导抗弓形虫感染的保护性研究(英文)[J]. 中国寄生虫学与寄生虫病杂志, 2010, 28(2): 81-88 |
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| 作者姓名： | 姚远 何深一 王花欣 周怀瑜 赵红 李婷 薛明福 朱兴全 |
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| 作者单位： | 1 山东大学医学院寄生虫学教研室，济南 250012；2 华南农业大学兽医学院寄生虫学教研室，广州 510642 |
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| 基金项目： | Supported by Shandong Provincial Natural Science Fund (No. ZR2009CM079); the Science and Technology Development Program of Shandong Province (No. 2006GG3202045); the Program for Changjiang Scholars and Innovative Research Team in Universities (No. IRT0723)~~ |
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| 摘 要： | 目的观察弓形虫SAG1-MIC8复合DNA基因疫苗对C57BL/6J小鼠的免疫保护作用。方法构建重组质粒pcDNA3.1-SAG1、pcDNA3.1-MIC8和pcDNA3.1-SAG1-MIC8,并分别转染Hela细胞体外表达蛋白,蛋白质印迹(Westernblotting)分析鉴定。将70只小鼠随机均分为5组,分别为PBS组、空质粒组、pcDNA3.1-SAG1组、pcDNA3.1-MIC8组和pcDNA3.1-SAG1-MIC8组,每2周肌注免疫(100μg/只)1次,共3次,于免疫前和初次免疫后13、27、41和55d采血分离血清。初次免疫后56d,每组小鼠中7只剖杀后分离脾细胞,另7只经腹膜感染弓形虫RH株速殖子(1×104/只),观察各组生存时间。ELISA法分别检测各组小鼠血清IgG、IgG1、IgG2b和IgG2c水平,以及干扰素γ(IFN-γ)和白介素4(IL-4)水平。放射法检测T淋巴细胞增殖情况。结果Westernblotting分析结果显示,重组质粒pcDNA3.1-SAG1、pcDNA3.1-MIC8和pcDNA3.1-SAG1-MIC8均能在Hela细胞表达,蛋白相对分子质量(Mr)分别为34000、74000和109000。pcDNA3.1-SAG1-MIC8组初次免疫后41d和55d血清中IgG,末次血清中IgG2b、IgG2c和IFN-γ,以及T淋巴细胞增殖能力均显著高于其他各组(均P0.05)。各组的末次血清中IgG1和IL-4水平差异均无统计学意义(均P0.05)。5组小鼠感染弓形虫速殖子后生存时间中位数分别为3、4、7、7和10d,各组间差异有统计学意义(P0.01)。结论弓形虫SAG1-MIC8复合DNA抗原较SAG1和MIC8单基因抗原有更好的免疫保护性。
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| 关 键 词： | 弓形虫；SAG1-MIC8复合抗原；核酸疫苗 |
Protective Immunity Induced in Mice by Multiantigenic DNA Vaccine with Genes Encoding SAG1 and MIC8 of Toxoplasma gondii |
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| YAO Yuan,HE Shen-yi,WANG Hua-xin,ZHOU Huai-yu,ZHAO Hong,LI Ting,XUE Ming-fu,ZHU Xing-quan. Protective Immunity Induced in Mice by Multiantigenic DNA Vaccine with Genes Encoding SAG1 and MIC8 of Toxoplasma gondii[J]. Chinese Journal of Parasitology and Parasitic Diseases, 2010, 28(2): 81-88 |
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| Authors: | YAO Yuan HE Shen-yi WANG Hua-xin ZHOU Huai-yu ZHAO Hong LI Ting XUE Ming-fu ZHU Xing-quan |
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| Affiliation: | YAO Yuan1,HE Shen-yi1,WANG Hua-xin1,ZHOU Huai-yu1,ZHAO Hong1,LI Ting1,XUE Ming-fu1,ZHU Xing-quan2 |
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| Abstract: | Objective To observe the immunoprotection induced by multiantigenic SAG1-MIC8 DNA vaccine of Toxoplasma gondii in C57BL/6J mice. Methods The sequences of genes encoding SAG1 and MIC8 protein were inserted into the eukaryotic expression vector pcDNA3.1 and the multiantigenic recombinant plasmid pcDNA3.1-SAG1- MIC8 was constructed. Then the recombinant plasmid was transfected into Hela cells to test its expression and the recombinant protein characterized by Western blotting. 70 mice were divided into 5 group... |
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| Keywords: | Toxoplasma gondii Multiantigenic SAG1-MIC8 DNA vaccine |
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