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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 investigate the dynamic changes of splenocyte proliferation, subpopulation and apoptosis after immunization of BALB / c mice with Schistosoma japonicum recombinant Bb (pGEX-Sj26GST-Sj32) vaccine. Methods 96 female BALB / c mice were randomly divided into two groups. The mice were immunized with rBb (pGEX-Sj26GST-Sj32) vaccine and oral inoculation (PO) and nasal mucosa (IN) Four mice in each group were sacrificed at 0, 2, 4, 6, 8, 10, 12, 14, 16, 18, 20 and 22 weeks. Spleens were harvested and spleen cells were isolated. Schistosoma japonicum adult antigen (SjAWA antigen ), And the proliferation of splenocytes in immunized mice was detected by MTT assay. At the same time, the control group consisted of ConA and control group. The splenic CD4 + and CD8 + T cell subsets of immunized mice were detected by flow cytometry (FCM) Splenocytes were stimulated with ConA, the incidence of apoptosis of splenocytes was detected by flow cytometry, and the original solution group was used as a control. Results The proliferation of PO group reached the peak at 4 weeks after immunization, while reached the peak at 12 weeks after immunization in IN group. The splenic CD4 + T lymphocyte subsets in PO group reached the highest level at 4 weeks after immunization, and the CD4 + T cell subsets The highest level was found at 12 weeks after immunization; the splenic CD8 + T lymphocyte subsets peaked at 10 weeks in PO group, and the highest level of CD8 + T cell subsets in IN group at 6 weeks after immunization; 6 weeks after immunization up to the highest, while the apoptotic rate of IN group spleen reached the highest level in 12 weeks. Conclusion The vaccine of Schistosoma japonicum recombinant Bb (pGEX-Sj26GST-Sj32) can induce the proliferation of splenocytes in early stage of immunity and induce the immune protection of CD4 + and CD8 + T cells in host and inhibit the apoptosis of splenocytes in mice.