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目的探讨日本血吸虫重组双歧杆菌属两歧双歧杆菌(Bifidobacterium bifidum,Bb)(pGEX-Sj14-3-3)疫苗免疫Balb/c小鼠后脾细胞因子的动态变化。方法重组疫苗分别经口服灌胃及鼻腔粘膜接种小鼠;在免疫后0、2、4、6、8、10、12、14、16、18、20和22周各剖杀4只小鼠,取脾及分离脾细胞;脾细胞分别经未刺激、SjAWA和ConA刺激培养;收集培养的上清液,双抗体夹心ELISA法测定培养上清液的细胞因子水平。结果口服组小鼠脾细胞IFN-γ、IL-12、TNF-α和IL-10水平分别在免疫后2~12、6~8、2~12和2~8周显著升高,分别在免疫后8、8、6和4周达最高水平;鼻腔黏膜接种组小鼠脾细胞IFN-γ、IL-12、TNF-α和IL-10水平分别在免疫后2~20、2~10、2~16和2~16周显著升高,分别在免疫后2、2、4和4周达最高水平。结论重组Bb(pGEX-Sj14-3-3)疫苗能够诱导免疫小鼠产生有效的免疫应答。
Objective To investigate the dynamic changes of splenic cytokines in Balb / c mice immunized with recombinant Bifidobacterium spp. Bifidobacterium bifidum (Bb) (pGEX-Sj14-3-3). Methods The recombinant vaccines were inoculated intragastrically and nasal mucosa by oral inoculation. Four mice were killed at 0,2,4,6,8,10,12,14,16,18,20 and 22 weeks after immunization. Spleen cells were isolated and spleen cells were isolated. Splenocytes were stimulated with unstimulated SjAWA and ConA respectively. The supernatants were harvested and the cytokine levels in the culture supernatants were determined by sandwich ELISA. Results The levels of IFN-γ, IL-12, TNF-α and IL-10 in spleen cells of mice in oral group were significantly increased at 2 ~ 12, 6 ~ 8, 2 ~ 12 and 2 ~ 8 weeks after immunization, After 8, 8, 6, and 4 weeks, the levels of IFN-γ, IL-12, TNF-α and IL-10 in spleen cells of nasal mucosal inoculated mice were respectively increased after 2 ~ ~ 16 and 2 ~ 16 weeks significantly increased, respectively, at 2, 4, and 4 weeks after immunization reached the highest level. Conclusion Recombinant Bb (pGEX-Sj14-3-3) vaccine can induce effective immune response in immunized mice.