摘要
目的分析重组弓形虫热休克蛋白70(rTgHSP70)鼻内或皮下免疫小鼠诱导的小肠黏膜IgA抗体分泌细胞(IgA antibody-secreting cells,IgA-ASCs)与小肠冲洗液sIgA抗体的动态变化及二者的相关性,探讨其上调小肠黏膜免疫应答的作用机制。方法 BALB/c小鼠90只,随机均分3组:鼻内组(20μg rTgHSP70/只,滴鼻,免疫2次,间隔2周)、皮下组(80μg rTgHSP70/只,背部皮下注射,免疫2次,间隔2周)、对照组(不免疫)。分别于末次免疫后第1、2、3、4、5、6周,每组取小鼠5只,脱颈椎处死,免疫组化法检测十二指肠、空肠及回肠黏膜IgA-ASCs数量,ELISA法检测小肠冲洗液sIgA水平。结果 IgA-ASCs分布于小肠黏膜的固有层中,且鼻内组小肠黏膜IgA-ASCs数量明显高于皮下组和对照组(P<0.001),鼻内组小肠冲洗液sIgA水平在免后1～4周均处于上升阶段,第4周达到高峰,显著高于皮下组和对照组,差异有统计学意义(P<0.001);鼻内组和皮下组小肠冲洗液sIgA水平与十二指肠、空肠、回肠黏膜固有层IgA-ASCs的数量呈正相关(P<0.05)。结论 rTgHSP70经鼻内或皮下免疫小鼠均可诱导小肠黏膜固有层IgA-ASCs和小肠冲洗液sIgA水平的持续高表达,且二者呈正相关,鼻内免疫上调小肠黏膜免疫的作用优于皮下免疫。
Objective To investigate the kinetics of IgA antibody-secreting cells（IgA-ASCs） in small intestinal mucosa of and specific sIgA level in small intestine washes of mice induced with recombinant Toxoplasma gondii heat shock protein 70 （rTgHSP70） by intranasal and subcutaneous immunizations, and analyze the correlation between IgA-ASCs and sIgA level as well as their roles in up-regulation of mucosal immune response in small intestine. Methods Ninety BALB / c mice were randomly divided into 3 groups. The mice in intranasal immunization group were immunized twice with 20 μg rTgHSP70 by intranasal drip at an interval of 2 weeks, and those in subcutaneous immunization group were injected s.c. with two times of 80 μg rTgHSP70 at an internal of 2 weeks, while those in control group were unimmunized. Five mice in each group were killed at weeks 1, 2, 3, 4, 5 and 6 after the last immunization respectively, of which the quantity of IgA- ASCs in mucosa of duodenum, jejunum and ileum were determined by immunohistochemical assay, and the sIgA level in intestinal washes by ELISA. Results The IgA-ASCs were found in lamina propria of small intestine mucosa. The quantity of IgA-ASCs in small intestine mucosa of mice immunized by intranasal drip was significantly higher than those in subcutaneous immunization and control groups （P 〈 0. 001 ）. The sIgA level in intestinal washes of mice increased 1 - 4 weeks and reached the maximum at week 4 after immunization by intranasal drip, which were significantly higher than those in subcutaneous immunization and control groups （P 〈 0. 001）. The sIgA level in intestinal washes of mice in intranasal and subcutaneous immunization groups were positively related to the quantity of IgA-ASCS in duodenum, jejunum and ileum mucosa lamina propria. Conclusion Immunization with rTgHSP70 by both intranasal and subcutaneous routes induced sustained high levels of IgA-ASCs in intestinal mucosal lamina propria and sIgA in intestinal washes. Positive relation was observed between the two levels. However, intranasal immunization approach is superior to subcutaneous injection in up-regulation of mucosal immunization in small intestine.
出处
《中国生物制品学杂志》
CAS
CSCD
2013年第2期191-195,共5页
Chinese Journal of Biologicals
基金
国家自然科学基金(81071374)
关键词
刚地弓形虫
热休克蛋白70
鼻内免疫
抗体分泌细胞
分泌性免疫球蛋白A
Toxoplasma gondii
Heat shock protein 70 （ HSP70 ）
Intranasal immunization
Antibody-secreting cells （ASCs）
Secretory immunoglobulin A（sIgA）
