弓形虫排泄-分泌抗原鼻内免疫小鼠诱导的脾与肠相关淋巴组织细胞免疫效应及动态变化

Dynamic of Cellular Immune Response of Spleen and Gut-associated Lymphoid Tissue Induced by Intranasal Immunization with Toxoplasma gondii Excreted-secreted Antigens in Mice

目的观察弓形虫与Vero细胞共培养提取的排泄-分泌抗原(ESA)鼻内免疫BALB/c小鼠诱导的脾及肠相关淋巴组织(GALT)细胞免疫效应及动态变化。方法84只BALB/c小鼠随机分为实验组和对照组,实验组以ESA(20μg/只)为抗原,对照组用未接种弓形虫的细胞培养上清液(20μl/只),鼻内免疫2次,间隔2周。末次滴鼻后第1、2、3、4、5、6、7周处死小鼠,分离脾、派伊尔结(PP)及小肠上皮内淋巴细胞(IEL)并计数。结果实验组小鼠脾淋巴细胞第3周达高峰,第1、2、3周明显高于对照组;PP淋巴细胞明显增生,第3周达高峰,第1、2、3、4、5周显著高于对照组;IEL第2周增生达高峰,第1～3周显著高于对照组。结论弓形虫ESA鼻内免疫BALB/c小鼠可有效诱导不同黏膜部位及系统特异性的细胞免疫应答,且可持续较长时间。

Objective To observe the dynamic of cellular immune response in spleen and gut-associated lymphoid tissue （GALT） of mice induced by intranasal immunization with Toxoplasma gondii RH strain excreted-secreted antigen（ESA） extracted from the co-culture of tachyzoites and Vero cells. Methods Divide 84 BALB/c mice into test and control groups randomly and immunize twice by intranasal drip, at an interval of 2 weeks, with 20 μg of ESA and 20 μg of Veto cell culture superatant containing no tachyzoites respectively. Kill a part of the mice 1,2,3,4,5,6 and 7 weeks after the 2nd immunization respectively for counting of lymphocytes in spleens and Peyer patches（PP） as well as intestinal intraepithehal lymphocytes（IEL）. Results The spleen lymphocyte counts of mice in test group reached a peak value 3 weeks, and were significantly higher than those in control group 1-3 weeks after the 2nd immunization. The counts of lymphocytes in PP of mice in test group reached a peak value 3 weeks, and were significantly higher than those in control group 1-5 weeks after the 2nd immunization. However,the IEL counts of mice in test group reached a peak value 2 weeks,and were significantly higher than those in control groups 1-3 weeks after the 2nd immunization. Conclusion Mucosal and systemic cellular immune responses were induced effectively and persistently by intranssal immunization with ESA of Toxoplasma gondii RH strain in mice.