弓形虫复合黏膜疫苗滴鼻免疫小鼠的黏膜相关淋巴细胞及其亚群的动态变化

Dynamic of Mucosal Associated Lymphocyte Subsets of Mice Immunized with Toxoplasma Complex Mucosal Vaccine by Intranasal Drip

目的探讨弓形虫复合黏膜疫苗滴鼻免疫小鼠的黏膜相关淋巴细胞及其亚群的动态变化。方法取BALB/c小鼠96只,随机分为实验组和对照组,实验组以弓形虫复合黏膜疫苗20μl/只滴鼻免疫,对照组以PBS滴鼻。滴鼻2次(间隔2周)后,分别于第1、2、3、4、6、8、10、12周处死小鼠,分离鼻相关淋巴组织(NALT)、Peyer结(PP)和上皮内淋巴细胞(IEL),制备淋巴细胞悬液,计数并涂片。免疫细胞化学法检测CD4+、CD8+T细胞亚群水平。结果实验组NALT、PP和IEL的T淋巴细胞均显著增生,PP和IEL的T淋巴细胞第2周达高峰,之后逐渐降低。与对照组相比,NALT的T淋巴细胞于第1、2、6、8、12周显著增生,PP的T淋巴细胞数于第1、2、3周显著增生,IEL于第1～4周显著增生。NALT和PP中主要以CD4+T细胞增生为主,肠IEL以CD8+T细胞增生为主。结论弓形虫复合黏膜疫苗滴鼻免疫BALB/c小鼠,可诱导鼻相关淋巴组织和肠相关淋巴组织T淋巴细胞明显增生,同时诱导其向不同亚群分化。

Objective To explore the dynamic of mucosal associated lymphocyte subsets of mice immunized with toxoplasma complex mucosal vaccine by intranasal drip. Methods Divide 96 BALB/c mice into test and control groups randomly. Each mouse in test group was immunized twice with 20μl of toxoplasma complex mucosal vaccine by intranasal drip at an interval of 2 weeks ,and that in control group with 20μl of PBS. Kill the mice at 1,2,3,4,6,8 and 10 weeks after immunization respectively, isolate nasal-associated lymphoid tissue（NALT） ,Peyer patches（PP） and intraepithelial lymphocytes（IEL） and prepare into suspension for lymphocyte counting and smearing. Determine the CD4^＋ and CD8^＋ T lymphocyte subsets by immunocytochemical method. Results Significant proliferations of T lymphocytes were observed in NALT, PP and IEL of mice in test group. The proliferations of lymphocytes in PP and IEL of test group reached a peak value at the 2nd week then decreased gradually. Compared with those of control group,the lymphocytes of test group proliferated significantly in NALT 1,2,6,8 and 12 weeks,in PP 1-3 weeks,and in IEL 1-4 weeks after immunization. The lymphocytes in NALT and PP were mainly CD4^＋ , and those in IEL were mainly CD8^＋. Conclusion The immunization with toxoplasma complex mucosal vaccine by intranasal drip induced significant lymphocyte proliferation in NALT,PP and IEL as well as the differentiation of lymphocytes to different subsets.