基础医学与临床 ›› 2014, Vol. 34 ›› Issue (1): 93-97.

• 研究论文 • 上一篇    下一篇

重组结核病疫苗rBCG-IL-12p70-TB10.4的免疫效应

张丰经,何红云,邓仪昊,杨开明   

  1. 大理学院
  • 收稿日期:2013-03-01 修回日期:2013-04-28 出版日期:2014-01-05 发布日期:2013-12-26
  • 通讯作者: 邓仪昊 E-mail:deng13032871868@163.com
  • 基金资助:
    大理学院博士科研启动基金

Immunization efficacy of a recombinant tuberculosis vaccine of rBCG-IL-12p70-TB10.4

  • Received:2013-03-01 Revised:2013-04-28 Online:2014-01-05 Published:2013-12-26
  • Contact: Yihao Deng E-mail:deng13032871868@163.com

摘要: 目的 将成功构建的重组卡介苗rBCG-IL-12p70-TB10.4免疫BALB/c小鼠,研究其免疫效应,为结核病新疫苗的研发奠定基础。方法 将融合基因IL-12p70-TB10.4克隆入pMV361构建重组质粒,以电转化方式将重组质粒导入卡介苗基因组,构建重组结核病疫苗rBCG-12p70-TB10.4 (rBCG-IT)。将此重组卡介苗免疫BALB/c小鼠,分别于免疫后3、6、9和12周,用间接ELISA检测特异性抗体滴度水平,XTT检测脾细胞增值反应, 流式细胞仪检测脾CD4+T、CD8+T细胞亚型的比例,ELISA试剂盒检测细胞因子的诱生。结果 在免疫后3~12周,rBCG-IT组脾细胞增殖反应均明显强于其它各组(P<0.01),该组IFN-γ诱生量及CD4+T细胞比例均显著高于rBCG-I组、rBCG-T组及BCG组(P<0.01)。IgG2a/IgG1比值在免疫后各时间点的变化趋势提示rBCG-IT可诱导Th1型细胞免疫应答。结论 重组疫苗rBCG-IT可诱导产生较BCG更强、持续时间更长的特异性细胞免疫应答,可对其免疫保护作用进行深入研究。

关键词: 重组卡介苗, IL-12p70, TB10.4, 免疫效应

Abstract: Objective The recombinant Bacilli Calmette-Guérin (BCG) of rBCG-IL-12p70-TB10.4 successfully constructed was used to vaccinate in BALB/c mice to investigate its immunogenicity, and to prepare groundwork in exploring new vaccines against Mycobacterium tuberculosis (MTB). Methods Fusion gene of IL-12p70-TB10.4 was cloned into multiple cloning sites of plasmid pMV361 to construct recombinant plasmid. The recombinant plasmid was transformed into BCG using Electroportation Generator to construct MTB vaccine of rBCG-12p70-TB10.4 (rBCG-IT) and then was immunized in Balb/c mice. 3, 6, 9, 12weeks after immunization, antibody titers were evaluated by indirect ELSIA, and cellular responses were evaluated by XTT, ratios of CD4+ T and CD8+ T cells were detected by flow cytometry, and IFN-γ production was evaluated by ELISA kit. Results The recombinant vaccine of rBCG-IT was able to induce high levels of specific antibody titers, stronger proliferative responses and greater IFN-γ production, compared with BCG vaccine. Conclusion The recombinant vaccine of rBCG-IT could elicit stronger and more long-lasting humoral and cell-mediated responses compared with BCG vaccine, and was worth exploring its protective efficacy.

Key words: Recombinant BCG, IL-12p70, TB10.4, immunization efficacy