霍乱毒素联合弓形虫排泄-分泌抗原鼻内免疫小鼠诱导的GALT和NALT黏膜免疫应答  

Intranasal immunization with cholera toxin and excreted-secreted antigens of Toxoplasma gondii induces an immune response at both NALT and GALT mucosal sites in mice

作　　者：李润花[1] 殷丽天[2] 孟晓丽[3] 刘红丽[3] 王海龙[3] 殷国荣[3] 

机构地区：[1]太原师范学院生物系,山西太原030031 [2]山西医科大学生理学系细胞生理学省部共建教育部重点实验室 [3]山西医科大学寄生虫学教研室

出　　处：《中国病原生物学杂志》2011年第4期265-268,259,共5页Journal of Pathogen Biology

基　　金：国家自然科学基金项目(No.81071374)

摘　　要：目的观察霍乱毒素（CT）联合弓形虫排泄-分泌抗原（ESA）鼻内免疫小鼠诱导的肠相关淋巴组织（GALT）和鼻相关淋巴组织（NALT）黏膜部位的免疫应答。方法将48只5～6周龄BABL／c小鼠随机均分为3组，分别用PBS、ESA和CT＋ESA滴鼻免疫小鼠2次，间隔2周。末次免疫后14d，处死小鼠，ELISA测定小鼠粪便和鼻咽冲洗液sIgA抗体水平；计数派伊尔淋巴集结（PP）、肠上皮淋巴细胞（IEL）、NALT和鼻通道（NC）淋巴细胞数，免疫细胞化学法检测CD4^＋、CD8^＋T细胞亚群水平。结果免疫后14d，CT＋ESA组小鼠鼻咽冲洗液和粪便sIgA水平显著高于ESA组和PBS组（P〈0．01）；CT＋ESA和ESA组小鼠GALT部位的PP淋巴细胞显著增生，主要以CD4^＋T细胞为主；而IEL以CD8^＋T细胞增生为主，CD4^＋／CD8^＋比值降低（P〈0．05）。CT＋ESA NALT内淋巴细胞明显增生。CT＋ESA组NC淋巴细胞显著升高（P〈0．01），以CD4^＋T细胞增生为主。结论CT佐剂联合ESA滴鼻免疫BALB／c小鼠可诱导GALT和NALT黏膜部位免疫应答。Objective To study the immune response at mucosal sites, i.e. gut-associated lymphoid tissue （GALT） and nasal associated lymphoid tissue （NALT）, after intranasal immunization with cholera toxin （CT） and excreted-secreted antigens （ESA） of Toxoplasma gondii. Methods BALB/c mice were randomly divided equally into three groups. Mice were intranasally immunized with PBS 20 μl, ESA 20 μg, or 1.0 μg CT ＋ 20μg ESA per mouse twice at an interval of two weeks. Mice were sacrificed on day 14 after final immunization. Levels of sIgA antibodies in feces and nasopharynx washes were detected by ELISA. Lymphocytes in Peyer＇s patches （PP）, intraepithelial lymphocytes （IEL）, NALT, and nasal passages （NC） were isolated and counted. The percentage of CD4^＋ and CD8^＋ T cells was determined by immunocytochemistry. Results The levels of sIgA antibodies in feces and nasopharynx washes of immunized mice in the CT＋ ESA group were higher than levels in PBS and ESA groups on day 14 after immunization （P〈0.05）. Lymphocytes in NALT increased significantly （P〈0.05） in the CT group after immunization. CD4^＋ and CD8^＋ T cell counts were higher than those of the PBS group （P〈0.01）, while the ratio of CD4^＋ and CD8^＋ T cells decreased （P〈0.05）. Lymphocytes in NC and PP increased （P〈0.01） with mainly CD4^＋ T cells increasing （P〈0.01）. With IEL, primarily the CD8＋ T count increased significantly （P〈0.01）, and the ratio of CD4^＋ and CD8^＋ T cells was significantly lower （P〈0. 05）. Conclusion This study indicated that intranasal immunization with cholera toxin and ESA of T. gondii can effectively induce immune response in both NALT and GALT.

关 键 词：刚地弓形虫 霍乱毒素 排泄-分泌抗原 黏膜免疫 肠相关淋巴组织 鼻相关淋巴组织 

分 类 号：R382.5[医药卫生—医学寄生虫学]