弓形虫ROP1基因真核表达重组质粒免疫小鼠后的免疫应答

中国寄生虫学与寄生虫病杂志 ›› 1999, Vol. 17 ›› Issue (6): 4-337.

弓形虫ROP1基因真核表达重组质粒免疫小鼠后的免疫应答

郭虹;陈观今;郑焕钦;周永安;吕芳丽

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

IMMUNE RESPONSES IN MICE VACCINATED WITH RECOMBINANT PLASMID pcDNA3 CONTAINING ROP1 GENE FROM TOXOPLASMA GONDII*

GUO Hong;CHEN Guanjin;ZHENG Huanqin;ZHOU Yongan;LU Fangli

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/Abstract

摘要： 　　目的：用构建的弓形虫ｐｃＤＮＡ３－ＲＯＰ１真核表达重组质粒，直接免疫小鼠，观察其所诱导的细胞及体液免疫反应。方法：碱裂解法大量制备ｐｃＤＮＡ３－ＲＯＰ１质粒，经肌肉注射免疫ＢＡＬＢ／ｃ小鼠，每鼠注射１００ μｇ，２ｗｋ后同量加强免疫１次，以ｐｃＤＮＡ３空质粒及生理盐水组为对照。分别于免疫后３０ｄ、５０ｄ、７０ｄ共３次用ＭＴＴ法测定小鼠脾脏Ｔ淋巴细胞增殖活性及ＮＫ细胞活性；用间接免疫荧光抗体法测定Ｔ淋巴细胞亚群数目；ＥＬＩＳＡ法测定ＩｇＧ抗体滴度。结果：用ｐｃＤＮＡ３－ＲＯＰ１质粒免疫小鼠３０ｄ后，脾脏明显增大；免疫组脾淋巴细胞增殖活性明显高于生理盐水及空质粒对照组。ＮＫ细胞杀伤活性３次的测定结果免疫组均高于对照组。Ｔ细胞亚群，ＣＤ４＋细胞数与对照组相比较无明显变化，而ＣＤ８＋细胞数显著增高。血清抗体ＩｇＧ７０ｄ内检测结果，与对照组相比较，无明显增高；而免疫后９０ｄ检测明显增高，抗体滴度１∶１００。结论：用ｐｃＤＮＡ３－ＲＯＰ１重组质粒ＤＮＡ免疫小鼠，可诱导产生细胞及体液免疫应答。

关键词: 弓形虫, 棒状体蛋白1, pcDNA3-ROP1重组质粒, DNA免疫

Abstract: 　AIM: To observe the immune responses in BALB/c mice vaccinated with the constructed recombinant plasmid, pcDNA3, containing ROP1 gene from Toxoplasma gondii. METHODS: The plasmid DNA obtained by alkaline lysis were injected into the left leg of each mouse at a dosage of 100 μg. A booster vaccine was given using the same dosage two weeks later. Control groups were injected with the pcDNA3 blank plasmid and normal saline, respectively. 30, 50 and 70 days after the booster injection, the proliferation activity of T lymphocytes and the NK cell activity were determined using MTT assay, the number of CD4+/CD8+ T cells with indirect immunofluorescence assay and the titer of IgG antibody by ELISA. RESULTS: The spleens of mice enlarged obviously after immunization with pcDNA3-ROP1. The activity of proliferation of spleen T lymphocytes was higher in the immunization group than in the control groups; The NK cell activity in the immunization group was higher than those of control ones. The number of CD4+ T cells showed no obvious increase but CD8+ T cells obviously increased. The titer of IgG antibody in the immunization group showed no significant increase until 90 days later. CONCLUSION: Vaccination of mice with the recombinant plasmid, pcDNA3-ROP1, could elicit cellular and humoral immune response in mice. \;

Key words: Toxoplasma gondii, rhoptry protein1(ROP1), pcDNA3-ROP1 recombinant plasmid, DNA vaccination

郭虹;陈观今;郑焕钦;周永安;吕芳丽. 弓形虫ROP1基因真核表达重组质粒免疫小鼠后的免疫应答[J]. 中国寄生虫学与寄生虫病杂志, 1999, 17(6): 4-337.

GUOHong;CHENGuanjin;ZHENGHuanqin;ZHOUYongan;LUFangli . IMMUNE RESPONSES IN MICE VACCINATED WITH RECOMBINANT PLASMID pcDNA3 CONTAINING ROP1 GENE FROM TOXOPLASMA GONDII*[J]. Chinese Journal of Parasitology and Parasitic Diseases, 1999, 17(6): 4-337.

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