日本血吸虫重组Bb(pGEX-Sj26GST-Sj32)疫苗免疫小鼠脾细胞增殖、亚群及凋亡的动态变化  

Dynamic changes of proliferation, subsets and apoptosis of splenocytes from BALB /c mice immunized with recombinant Bb( pGEX-Sj26GST-Sj32 ) vaccine against Schistosoma japonicum

作　　者：向进平[1] 李文桂[1] 张丽[1] 

机构地区：[1]重庆医科大学附属第一医院传染病寄生虫病研究所,重庆400016

出　　处：《细胞与分子免疫学杂志》2013年第11期1129-1132,共4页Chinese Journal of Cellular and Molecular Immunology

基　　金：重庆市科委地方病重大专项基金项目(2008AB5055;2008AB5008;2008AB5054)

摘　　要：目的探讨日本血吸虫重组Bb(pGEX-Sj26GST-Sj32)疫苗免疫BALB/c小鼠后脾细胞增殖、亚群及凋亡的动态变化。方法 96只雌性BALB/c小鼠随机均分为2组,用rBb(pGEX-Sj26GST-Sj32)疫苗分别经口服灌胃(PO)及鼻腔黏膜(IN)接种两种途径免疫小鼠,于免疫后0、2、4、6、8、10、12、14、16、18、20和22周每组各剖杀4只小鼠,取脾,分离脾细胞,用日本血吸虫成虫抗原(SjAWA抗原)刺激培养,MTT法检测免疫小鼠脾细胞增殖反应,同时设原液、刀豆蛋白A(ConA)组对照;用流式细胞术(FCM)检测免疫小鼠脾CD4+和CD8+T细胞亚群的百分比;脾细胞经ConA刺激培养,流式细胞仪检测脾细胞的凋亡发生率,并设原液组作为对照。结果 PO组增殖水平于免疫后4周达高峰,而IN组于免疫后12周达高峰;PO组脾CD4+T细胞亚群于免疫后4周达最高水平,IN组CD4+T细胞亚群于免疫后12周达最高水平;PO组脾CD8+T细胞亚群于10周达峰值,IN组CD8+T细胞亚群于免疫后6周达最高水平;PO组小鼠脾细胞凋亡率于免疫后6周达最高,而IN组脾细胞凋亡率于12周达最高水平。结论日本血吸虫重组Bb(pGEX-Sj26GST-Sj32)疫苗在免疫早期可引起脾细胞增殖,诱导CD4+和CD8+T细胞参与宿主的免疫保护,抑制小鼠脾细胞的凋亡。Objective To study the dynamic change of proliferation, subsets and apoptosis of splenocytes of BALB/c mice immunized with recombinant Bb （pGEX-Sj26GST-Sj32） vaccine of Schistosoma japonicum. Methods A total of 96 BALB/c mice were randomly divided into 2 groups （n = 48 per group） and immunized with the recombinant vaccine orally （ PO group） and intranasally （IN group） respectively. Then autopsies were made in 4 mice of each group 0, 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22 weeks after immunization to separate splenocytes. After being cultured under the activation of SjAWA, the splenocytes were observed in the proliferative response using the MTT assay. The stock （non-stimulation） and concanavalin A （ConA） activation were used as controls; The proportions of CD4＋ and CD8＋ T cells were determined by flow cytometry （FCM） ; Splenocytes cultured under the activation of ConA were collected to detect the apoptotic rates of splenocytes by FCM, with the stock group as a control. Results The level of splenocyte proliferation in PO group peaked on week 4 after immunization, while the level in IN group did so on week 12. The level of CD4＋T cell subsets in PO group reached the highest on week 4 after immunization, while the level in IN group did so on week 12. The level of CD8＋ T cell subsets in PO group rose to the top on week l0 after immunization, while the level in IN group did so on week 6. The apoptotic rate of splenocytes in PO group was the highest on week 6 after immunization, while the rate in IN group did so on week 12. Conclusion The recombinant Bb（ pGEX- Sj26GST-Sj32） vaccine could cause proliferation of splenocytes, induce immune protection to the host by up-regulating CD4＋ and CD8 ＋ T cells and inhibit the apoptosis of splenocytes from mice.

关 键 词：脾细胞增殖 脾细胞亚群 凋亡 日本血吸虫 疫苗 

分 类 号：R392.12[医药卫生—免疫学] Q255[医药卫生—基础医学]