重组弓形虫SAG1蛋白免疫小鼠抗弓形虫攻击保护作用  

Protection in mice immunized with recombinant expressing TgSAG1 against Toxoplasma gondii challenge

作　　者：葛玉芳[1,2] 张建伟[1,2] 殷国荣[1] 刘晋平[2] 孟晓丽[1] 刘美娜[1] 曹蕾[1] 

机构地区：[1]山西医科大学医学寄生虫学研究所,寄生虫学教研室,太原030001 [2]山西农业大学动物科技学院

出　　处：《山西医科大学学报》2009年第9期804-807,共4页Journal of Shanxi Medical University

基　　金：国家自然科学基金资助项目(30640057);山西医科大学青年基金资助项目

摘　　要：目的评价重组弓形虫表面抗原1(rTgSAG1)蛋白皮下免疫小鼠诱导的免疫应答及抗弓形虫攻击的保护作用。方法6周龄BALB/c小鼠60只,随机分为5组,分别以100μlPBS、20μg、40μg、60μg或80μg rTgSAG1(溶于100μlPBS)/只皮下注射免疫小鼠3次,间隔14d。末次免疫后14d,以RH株弓形虫速殖子1×104个/只灌胃攻击所有小鼠,每日观察小鼠健康状况。攻虫后第28天,处死小鼠。分离并计数肝和脑组织内速殖子,分离并计数脾淋巴细胞和小肠上皮内淋巴细胞(IEL),ELISA法检测血清IgG和小肠冲洗液IgA抗体水平。结果攻虫后6-14d,小鼠出现不同程度的被毛粗糙、采食饮水减少,PBS组存活率最低(33.3%),40μg组最高(75%)。40μg组肝(79.60)和脑组织内荷虫数(16.88)低于PBS组(分别为94.43和19.48)(均P<0.05),脾淋巴细胞数(10.31)和IEL数(4.25)高于PBS组(6.24,1.64)(均P<0.01);40μg组(P<0.01)、20μg组和60μg组(P<0.05)血清IgG抗体水平显著高于PBS组;各组肠液IgA抗体水平无明显变化。结论rTgSAG1蛋白皮下免疫小鼠能诱导产生抗弓形虫感染保护性免疫,40μg为免疫的最适剂量。Objective To evaluate the protective effect against Toxoplasma gondii in mouse hypodermic immunization with recombination Toxoplasma gondii SAG1 （rTgSAG1） protein. Methods Sixty 6-week-old BALB/c mice were randomly divided into five groups, 12 mice per group. Mice were hypodermically immunized with 100 μl PBS,20 μg, 40 μg,60 μg and 80 μg rTgSAG1 （dissolved in 100 μl PBS） ,respectively. Two boosters were carried out at 2-week intervals. On day 14 after the last immunization,all mice were intragastrically challenged with 1 × 104 tachyzoites per mouse, and the health status of infected mice was recorded every day after challenged. On day 28 after challenge, survival mice were killed. The tachyzoites of their livers and brains and the lymphocytes of spleens and intraepithelial lymphocytes （IEL）were counted, respectively, and serum IgG and IgA in intestinal washes were detected by ELISA. Results On day 6 - 14 after challenge, mice showed clothing hair rough at different degrees, decrease of foraging and drinking water. The survival rate was the lowest in PBS group（ 33.3% ）, but the highest（75% ） in 40 μg dose group. The tachyzoite loads in 40 μg dose group in livers （79.60） and brains （ 16.88 ） were significantly lower than that in PBS group （ 94.43,19.48 ） （ P 〈 0.05 ）. The number of splenic lymphocyte （ 10.31 ） and IEL （4.25） in 40 μg group were significantly higher than that in PBS group （ 6.24,1.64 ） （ P 〈 0.01 ）. The levels of IgG in 40 p,g group （P 〈 0.01 ） ,20 p,g and 60 μg groups （P 〈 0.05 ）were significantly higher than that in PBS group respectively. The level of IgA had no marked change. Conclusion The protective immunity against Toxoplasma gondii in mouse hypodermic immunization with rTgSAG1 could be induced,and 40 μg is the best immunizing dose.

关 键 词：刚地弓形虫 表面抗原SAG1 重组蛋白 保护性免疫 

分 类 号：R382.5[医药卫生—医学寄生虫学]