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目的 观察弓形虫主要表面抗原SAG1、SAG2 与霍乱毒素A2 /B亚基复合基因真核质粒经肌肉免疫小鼠所诱导的免疫反应。方法 将SAG1基因、SAG2 基因及CTXA2 /B基因定向连接插入真核表达质粒pcDNA3.1,经酶切及测序,获得pcDNA3.1 SAG1 SAG2 及pcDNA3.1 SAG1 SAG2 CTXA2 /B的重组子;碱裂解法大量制备经肌肉注射免疫BALB/c鼠,每只鼠经后腿肌肉注射质粒10 0 μg ,每2周免疫1次,共3次,以PcDNA3.1空质粒注射组及PBS组为对照,分别于每次免疫前断尾取血和免疫后4周取小鼠脾脏测定T淋巴细胞增殖活性及NK细胞活性,ELISA法测定IgG抗体。结果 免疫组小鼠的IgG抗体水平明显提高,NK细胞杀伤活性和T细胞增殖活性也明显增强。免疫鼠抗攻击感染的时间延长。结论 含有霍乱毒素的复合基因免疫小鼠后体液免疫和细胞免疫水平均有提高。
Objective To observe the immune responses induced by intramuscular immunization of eukaryotic plasmids containing the major surface antigens SAG1, SAG2 and cholera toxin A2 / B subunit of Toxoplasma gondii. Methods The SAG1 gene, SAG2 gene and CTXA2 / B gene were inserted into the eukaryotic expression plasmid pcDNA3.1. After digestion and sequencing, recombinant plasmids of pcDNA3.1 SAG1 SAG2 and pcDNA3.1 SAG1 SAG2 CTXA2 / B were obtained. Method A large number of BALB / c mice were immunized intramuscularly by intramuscular injection. Each mouse was intramuscularly injected with 100 μg plasmid into the hind leg and immunized once every 2 weeks for 3 times. PcDNA3.1 empty plasmid injection group and PBS group were used as controls. T lymphocyte proliferation activity and NK cell activity were measured at 4 weeks after the mice were pruned and immunized respectively before immunization. IgG antibodies were detected by ELISA. Results The level of IgG antibody in immunized mice was significantly increased, and the cytotoxic activity of NK cells and T cell proliferative activity were also significantly increased. Immune mouse anti-attack infection prolonged. Conclusion The humoral and cellular immunity of mice immunized with the cholera toxin-containing complex gene were both improved.