目的:动态观察多房棘球绦虫重组BCG-Em14-3-3疫苗免疫小鼠后脾细胞因子的变化。方法:将疫苗采用鼻腔内接种免疫BALB/C鼠,在免疫后0、2、4、6、8、10、12、14、16和18周各剖杀4只小鼠取脾,分离脾细胞,用EmAg或ConA刺激培养,收集脾细胞培养上清液,用试剂盒检测脾细胞培养上清液的IL-2、IFN-γ、TNF-α和IL-4水平,同时设有PBS对照。结果:疫苗接种组的IL-2、IFN-γ、TNF-α和IL-4分别在免疫后4~8周、2~8周、2~8周和6~10周升高,分别在免疫后6、6、4~6和8周达最高水平。结论:多房棘球绦虫重组BCG-Em14-3-3疫苗在免疫早期(2~8周)即可诱导小鼠产生一个TH1型保护性免疫反应。
Objective:To dynamically observe the changes in cytokines of splenocytes in mice immunized with recombinant BCG-Em14-3-3 vaccine of Echinococcus multilocularis (Em). Methods: BALB/c mice receiving intranasal immunization with recombinant BCG-Em14-3-3 vaccine were killed and the spleens were sampled on weeks 0,2,4,6,8,10,12,14,16,and 18 of the immunization. The splenocytes were isolated and cultured under the stimulation of EmAg or ConA. The supernatants were collected ,and the levels of interleukin-2 (IL-2),IL-4,interferon-γ (IFN-γ),and tumor necrosis factor-α (TNF-α) were determined by enzyme-linked immunosorbent assay Kit,with PBS as the control. Results: In the immunization groups,the levels of IL-2,IFN-γ,TNF-α,and IL-4 increased from week 4 to 8,week 2 to 8,week 2 to 8,and week 6 to 10 and reached the peak on week 6,week 6,week 4 to 6,and week 8,respectively. Conclusion: TH1 response is induced in mice by rBCG-Em14-3-3 vaccine at the early stage of immunization (2 to 8 weeks).