南方医科大学学报

南方医科大学学报 ›› 2015, Vol. 35 ›› Issue (02): 202-.

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日本血吸虫重组Bb（pGEX-Sj32）疫苗诱导BALB/c鼠脾细胞增殖、亚群及细胞因子的动态变化

谭建蓉，李文桂，覃婷   

  • 出版日期:2015-02-20 发布日期:2015-02-20

Changes in splenocyte proliferation, subsets and cytokine production in mice immunized
with recombinant vaccine Bifidobacterium bifidum (pGEX-Sj32) of Schistosoma japonicum

  • Online:2015-02-20 Published:2015-02-20

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摘要： 目的观察日本血吸虫重组Bb（pGEX-Sj32）疫苗诱导免疫鼠脾细胞免疫应答的动态变化，探讨该重组疫苗的免疫机制。
方法88只BALB/c鼠随机分为口服组和滴鼻组，口服组小鼠经口服单次接种106克隆形成单位（CFU）重组疫苗，滴鼻组小鼠经
滴鼻单次接种105 CFU重组疫苗；于免疫后0~20周内间隔2周每组随机剖杀4只小鼠，取脾并制备脾细胞悬液，分别经未刺激
（脾细胞悬液）、SjAWA刺激和ConA刺激培养。MTT法检测脾细胞增殖效率，流式细胞术检测脾CD4+T细胞和CD8+T细胞亚
群变化，双抗体夹心ELISA法检测培养上清液中TNF-α、IL-10 和IL-12 表达水平。结果与0 周相比，未刺激、SjAWA刺激和
ConA刺激时，口服组和滴鼻组脾细胞均分别在免疫后2~16周和2~18周发生极显著增殖（P<0.01）；两组CD4+T细胞亚群均于
免疫后2~12周显著升高（P<0.01），CD8+T细胞亚群在观察期间升高不明显；不同培养条件下，口服组脾细胞因子TNF-α、IL-10
和IL-12水平均分别在免疫后2~14周、2~18周和2~14周升高，于8周、10周和6周达峰值；滴鼻组3者分别在免疫后2~14周、2~
18周和2~18周升高，于8周、10周和8周达峰值。结论日本血吸虫重组Bb（pGEX-32）疫苗可诱导小鼠产生有效的免疫应答。

Abstract: Objective To observe the dynamic changes of immune responses of splenocytes in mice immunized with
recombinant vaccine Bifidobacterium bifidum (pGEX-Sj32) of Schistosoma japonicum and investigate the immunological
mechanism of the vaccine. Methods Eighty-eight BALB/c mice were randomized for immunization with 106 CFU recombinant
vaccine orally or with 105 CFU recombinant vaccine intranasally. Four mice were selected from each group every two weeks to
test the responses of the splenocytes to stimulations with SjAWA or ConA. MTT assay and flow cytometry were used to assess
splenocyte proliferation and the distribution of CD4+ and CD8+ T cells, respectively; the levels of interleukin-10 (IL-10), IL-12
and tumor necrosis factor-α (TNF-α) in the cell culture supernatant were detected by ELISA. Results Regardless of the
stimulations, the splencytes showed significantly enhanced proliferation in weeks 2-16 in oral administration group and in
weeks 2-18 in intranasal group (P<0.01). CD4+ subsets in both two groups increased obviously in weeks 2-12 (P<0.01) but CD8+
subsets remained stable. In oral administration group, the levels of TNF-α, IL-10 and IL-12 increased in weeks 2-14, 2-18 and
2-14, and peaked at week 8, 10 and 6, respectively; in intranasal group, the cytokines increased in weeks 2-14, 2-18 and 2-18,
and peaked at week 8, 10 and 8, respectively. Conclusion The recombinant vaccine rBb (pGEX-Sj32) can induce effective
immune responses in mice.