小鼠腹水中弓形虫排泄分泌抗原鼻内免疫小鼠诱导的免疫应答  

Immune responses induced by intranasal immunization with Toxoplasma gondii excreted/secreted antigens extracted from peritoneal fluid of infected mice

作　　者：刘娟娟[1] 殷国荣[1] 管志玉[1] 石蓉[1] 张宇斌[1] 孟晓丽[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》2007年第2期110-114,共5页Journal of Pathogen Biology

基　　金：国家自然科学基金资助项目(No.30640057);山西省自然科学基金项目(No.20041105);山西省高校科技研究开发重点项目(No.20041238)

摘　　要：目的观察弓形虫感染小鼠腹水中分离的排泄-分泌抗原（excreted/secreted antigens,ESA）鼻内免疫小鼠对不同粘膜部位和系统的免疫效应。方法将5～6周龄BALB/c小鼠随机分为4组,每组10只,实验组分别用从感染弓形虫后24、48、72 h小鼠腹水中提取的ESA 20μg/只鼻内免疫小鼠2次,间隔2周;对照组用20μl/只PBS滴鼻。观察小鼠健康和死亡情况,记录体重。末次免疫后第3周处死小鼠,计数PP（Peyer’s patches）个数;分离PP淋巴细胞、脾淋巴细胞、小肠上皮内淋巴细胞（intraepithelial lymphocyte,IEL）及肠系膜淋巴结细胞（mesenteric lymph node lympho-cyte,MLNL）并计数。眼眶采血和收集直肠内粪便,ELISA检测血清特异性IgG水平及粪便sIgA。结果72 h ESA组小鼠免疫后健康状况较差,体重逐渐降低,死亡4只;其他组小鼠体重仍呈增高趋势。各实验组小鼠IEL、MLNL、脾淋巴细胞以及PP淋巴细胞的增殖活性均高于对照组（P〈0.05）,72 h和48 h ESA组高于24 h ESA组（P〈0.01）。免疫后第3周,各实验组小鼠血清IgG、粪便sIgA水平均高于对照组（P〈0.05）,72 h和48 h ESA组高于24 h ESA组（P〈0.01）。结论感染后不同时间提取的腹水弓形虫ESA鼻内免疫小鼠均可诱导不同粘膜部位及系统特异性的免疫应答,48 h ESA的免疫效果最好。Objective To investigate the mucosal and systemic immune responses after intranasal immunization with Toxoplasrna gondii RH strain excreted/secreted antigens （ESA） extracted from the peritoneal fluids of infected mice and determine the optimal antigens group of ESA. Methods Forty 5- to 6-weeks-old BALB/c mice were randomly divided into four groups, each group were intranasally immunized two times at 14-day intervals. Three groups received with 24 h, 48 h, 72 h ESA and 20 μg per mouse, while mice intranasally administrated with PBS were control. The condition of mice about health and death was observed and the weight of mice was recorded every day. Mice were killed on the third week after the last immunization. Serum IgG and sIgA in feces were detected by ELISA. The number of the Peyer＇s patches was counted. Lymphocytes in PP, spleen, IEL, MLN were isolated and counted. Results The weight of 72 h group mice gradually decreased after the first immunization, while that of other groups mice still remained increasing （P〈0.05）. 48 h group and 72 h group, the lymphocytes in spleens, MLN, PP and IEL was significantly higher than that of control group （P〈0.01）. The level of IgG and sIgA in all immunized mice was higher than that of control （P〈0.05）, Conclusion Intranasal immunization with ESA from the infected mice peritoneal fluids can effectively induce the mucosal and systemic immune responses, the ESA of 48 h is optimal antigens.

关 键 词：弓形虫 排泄-分泌抗原(ESA) 粘膜免疫 鼻内免疫 粘膜疫苗 

分 类 号：R382.5[医药卫生—医学寄生虫学]