不同剂量弓形虫ESA鼻内免疫小鼠诱导的粘膜及系统免疫应答  

Intranasal immunization with different-dosage Toxoplasma gondii ESA induces mucosal and systemic immune responses in mice

作　　者：申金雁[1] 管志玉[1] 刘红丽[1] 孟晓丽[1] 王海龙[1] 殷国荣[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,山西太原030001

出　　处：《中国病原生物学杂志》2008年第3期186-189,共4页Journal of Pathogen Biology

基　　金：国家自然科学基金项目(No30640057);山西省自然科学基金项目(No20041105);山西省高校科技研究开发重点项目(No20041238);山西省青年科技基金项目(No20051045)

摘　　要：目的观察不同剂量弓形虫排泄-分泌抗原(excreted/secreted antigens,ESA)鼻内免疫小鼠诱导的粘膜及系统免疫应答,确定适宜免疫剂量。方法40只BALB/c小鼠,雌雄各半,随机分为5组,每组8只。分别用5、10、20和30μgESA滴鼻免疫小鼠2次,间隔14d,对照组用20μl/只PBS滴鼻。于末次免疫后第14d,颈椎脱臼处死小鼠,ELISA法检测鼻咽冲洗液和小肠冲洗液sIgA、血清IgG抗体水平,分离并计数肠上皮内淋巴细胞(intestinal intraepitheliallym-phocytes,iIEL)、肠系膜淋巴结淋巴细胞(mesenteric lymph node lymphocytes,MLNL)及脾淋巴细胞。结果实验期间,小鼠健康状况良好。随鼻内免疫ESA剂量的增加,特异性抗体水平不同程度的升高,MLNL、iIEL及脾淋巴细胞也显示不同程度的增生性应答。其中,30μg组小鼠血清IgG、脾淋巴细胞数量与10μg组、5μg组和PBS组比较,差异有统计学意义(P<0.05或P<0.01);30μg组小肠冲洗液sIgA、鼻咽冲洗液sIgA、iIEL和MLNL数量,20μg组小肠冲洗液sIgA和MLNL数量与5μg组及PBS组比较,差异有统计学意义(P<0.05或P<0.01);20μg组血清IgG与PBS组比较差异有统计学意义(P<0.05)。结论20μg和30μgESA鼻内免疫能有效诱导粘膜及系统免疫应答。Objective With the aim of determining the optimal dosage of excreted-secreted antigens （ESA）, we observed that different-dosage ESA administrated intranasally induced mueosal and systemic immune responses. Methods For immunization of the mice, 40 mice were allocated into 5 groups, the groups were immunized intranasally with 5 μg ESA, 10 μg ESA, 20 μg ESA and 30 μg ESA per mouse, respectively, the control group with 20 μl PBS-immunized. All mice were immunized intranasally twice at interval of 14 days. All mice were killed on day 14 after the last immunization. sIgA in nasopharyngeal and intestinal washes and IgG in serum were detected by ELISA. Intestinal intraepithelial lymphoeytes （iIEL）, mesenteric lymph node lymphocytes （MLNL） and spleen lymphoeytes were isolated and counted. Resuits At the whole experimental session, all mice were healthy. With the increase of dosage of ESA, the level of antibodies and number of iIEL, MLNL and spleen cell in experimental groups were rising, compared with PBS group. In 30μg group, the level of IgG in serum and number of spleen cell was higher significantly than in 10 μg, 5 μg and PBS group （P 〈0.05 or P〈0.01）. The level of sIgA in intestinal washes, that in nasopharyngeal washes （P〈0.05）, the number of iIEL and MLNL in 30 ttg group, the level of sIgA in intestinal washes and the number of MLNL in 20μg group were higher than that in PBS group and 5 μg group （P〈0.01 or P〈0.05）. The level of IgG in serum in 20 μg group was only higher than in PBS group （P〈0.05）. Conclusion 30 μg or 20 μg ESA had induced mueosal and systemic immune response.

关 键 词：弓形虫 排泄分泌抗原 鼻内免疫 粘膜疫苗 

分 类 号：R382.5[医药卫生—医学寄生虫学]