日本血吸虫重组Bb(pGEX-Sj14-3-3)疫苗免疫小鼠脾细胞因子的动态观察

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。

Dynamic observation on splenocyte cytokines in mice immunized with recombinant Bb(pGEX-Sj14-3-3) vaccine against Schistosoma japonicum

To explore the availability of the recombinant Bb(pGEX-Sj14-3-3) vaccine,we investigated the dynamic changes of cytokines in splenocyte supernatants from Balb/c mice immunized with the vaccine.Mice were immunized with the recombinant vaccine orally and intranasally respectively,and then sacrificed at different time points of week 0,2,4,6,8,10,12,14,16,18,20 and 22 after immunization.Spleens were separated,and splenocytes were cultured with or without SjAWA or ConA stimulation respectively.The splenocyte supernatants were collected to determine the levels of IFN-γ,IL-12,TNF-α and IL-10 by sandwich ABC-ELISA.We found that the levels of IFN-γ,IL-12,TNF-α and IL-10 in the oral group significantly was increased on 2-12th,6-8th,2-12th and 2-8th weeks respectively,reached to peaks on week 8,8,6 and 4 respectively.The levels of IFN-γ,IL-12,TNF-α and IL-10 in the intranasal group predominantly increased on 2-20 th,2-10 th,2-16th and 2-16th weeks respectively,reached to the highest levels on week 2,2,4 and 4 respectively.In conclusion,the rBb(pGEX-Sj14-3-3) vaccine could induce effective immune response in inoculated mice,which provide valuable information on the exploitation and utilization of this recombinant vaccine.