细粒棘球绦虫重组BCG-Eg95疫苗免疫小鼠后脾细胞亚群的动态观察

摘要

Objective To dynamically observe the changes of subsets of splenocytes in mice by immunization with recombinant BCG-Eg95 vaccine of Echinococcus granulosus(Eg).Methods Balb/c mice were divided randomly into 3 groups according to their weishts:intranasal group.per os group and PBS control.The mice were vaccinated intranasally or orally by the vaccine respectively in experimental groups.and the control mice were given phosphate buffer saline intranasally.These mice were killed to get spleen on 0,2,4,6,8,10,12, 14,16 and 18 week of immunization,respectively.Splenocytes were separated to measure subsets of CD4+ and CD8+T ceUs by FACsort.Results There were marked differences in ratio of CD4+ and CD8+ subsets among the different groups(F value were 21.56 and 22.08 respectively,P＜0.05).There were very marked differences in ratio of CD4+ and CD8+ subsets in different weeks(F value were 5.75 and 6.29 respectively.P＜0.01).In the intranasal group,CD4+ and CD8+ T subsets increased obviously in 6～18 weeks and 12 weeks,and reached the highest level on 10 and 12 week,espectively.Their values were 0.348±0.013 and 0.090±0.003.respectively.There were marked or very marked differences(q value were 7.32 and 5.32 respectively,P＜0.01 or＜0.05)in comparison with 0 week(0.230±0.022 and 0.069±0.015).In the oral group,CD4+and CD8+ subsets rose reinarkablv on 6-16 weeks and 8-18 weeks,achieved the hishest level on 10 and 16 weeks,respectively.Their vahes were 0.405± 0.006 and 0.096±0.004,respectively.There were marked or very marked difference(q value were 7.53 and 5.35 respectively,P＜0.01 or＜0.05)in comparison with week 0(0.230±0.022 and 0.069±0.015).Conclusion CD4+and CD8+T subsets may play an important role in immune response induced in mice by rBCG-Eg95 vaccine.%目的 动态观察细粒棘球绦虫重组BCG-Eg95疫苗免疫小鼠后脾细胞亚群的变化.方法 将120只Balb/c小鼠按体质量随机分为3组:鼻腔内接种组、口服灌胃组、对照组.将疫苗分别采用鼻腔内接种和口服灌胃免疫实验组小鼠,对照组单次接种10μl磷酸盐缓冲液(PBS)于小鼠鼻腔黏膜.在免疫后0、2、4、6、8、10、12、14、16、18周各剖杀4只小鼠,取脾脏,分离脾细胞,流式细胞仪检测脾CD4+、CD8+T淋巴细胞亚群的百分比.结果 不同组的小鼠脾细胞CD4+、CD8+亚群百分比比较差异有统计学意义(F值分别为21.56、22.08,P＜0.05),不同周数的小鼠脾细胞CD4+、CD8+亚群百分比比较差异有统计学意义(F值分别为5.75、6.29.P＜0.01).鼻腔内接种组的脾CD4+、CD8+T细胞亚群分别在免疫后6-18、12周升高,在免疫后10、12周达最高水平,其值分别为0.348±0.013、0.090±0.003.与0周(0.230±0.022、0.069±0.015)比较差异有统计学意义(q值分别为7.32、5.32,P＜0.01或＜0.05);口服灌胃组脾细胞CD4+、CD8+T细胞亚群分别在免疫后6～16、8-18周增加,分别在免疫后10、16周达最高水平,其值分别为0.405±0.006、0.096±0.004,与0周(0.230±0.022、0.069±0.015)比较差异有统计学意义(q值分别为7.53、5.35,P＜0.01或＜0.05).结论 CD4+、CD8+T细胞亚群在细粒棘球绦虫重组BCG-Eg95疫苗诱导的小鼠免疫应答中起重要作用.