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目的观察重组弓形虫过氧化物还原蛋白(rTgPrx)滴鼻免疫小鼠诱导的免疫应答及抗弓形虫感染作用。方法6周龄BALB/c小鼠75只,随机分为5组,实验组分别用10,20,30,40μg rTgPrx〔溶于20μL磷酸盐缓冲液(PBS)中〕滴鼻免疫小鼠3次,各间隔2周;对照组用等量磷酸盐缓冲液;末次免疫后14 d,用1×104个速殖子/只灌胃攻击全部小鼠;攻击后30 d,眼静脉丛采血,颈椎脱臼处死小鼠,ELISA检测血清IgG和肠液IgA,分离并计数肠上皮内淋巴细胞(IEL)和脾淋巴细胞,分离并计数肝、脑组织内弓形虫速殖子。结果40μg rTgPrx组血清IgG、IEL及脾淋巴细胞数量均高于对照组(P<0.05),40μg组肝、脑组织内虫荷(速殖子数)明显低于对照组及10,20和30μg组(P<0.05)。结论rTgPrx滴鼻免疫小鼠可诱导黏膜部位和系统免疫应答和抗弓形虫感染作用,40μg组的免疫保护作用优于其他剂量组。
Objective To observe the immune response induced by intranasal immunization with recombinant Toxoplasma gondii (rTgPrx) and its anti-Toxoplasma infection effect. Methods 75 BALB / c mice of 6 weeks old were randomly divided into 5 groups. In the experimental group, mice were immunized intranasally with 10, 20, 30 and 40 μg rTgPrx [dissolved in 20 μL phosphate buffered saline (PBS) , The rats in control group were treated with equal volume of phosphate buffered saline (PBS); all the mice were challenged with 1 × 104 tachyzoites / gavage 14 days after the last immunization; 30 days after challenge, ophthalmic Mice were sacrificed by dislocation, serum IgG and intestinal IgA were detected by ELISA, intestinal epithelial lymphocytes (IELs) and splenic lymphocytes were isolated and counted, Toxoplasma gondii tachyzoites were isolated and counted in liver and brain. Results The serum levels of IgG, IEL and splenic lymphocytes in 40 μg rTgPrx group were significantly higher than those in control group (P <0.05). The numbers of tachyzoites (tachyzoites) in liver and brain of 40 μg group were significantly lower than those in control group and 10, 20 and 30 μg Group (P <0.05). Conclusion The mice immunized with rTgPrx intranasally can induce mucosal site, systemic immune response and anti-Toxoplasma infection. The immunoprotection of 40μg group is superior to other dose groups.