基础医学与临床 ›› 2011, Vol. 31 ›› Issue (7): 746-750.

• 研究论文 • 上一篇    下一篇

约氏疟原虫MIF分子调节巨噬细胞炎性因子基因表达

邵丁丁1,曾山1,钟翔2,王恒1   

  1. 1. 中国医学科学院基础医学研究所
    2. 中国医学科学院基础医学研究所&北京协和医学院基础学院
  • 收稿日期:2011-03-28 修回日期:2011-05-10 出版日期:2011-07-05 发布日期:2011-07-05
  • 通讯作者: 王恒 E-mail:wanghpumc@gmail.com
  • 基金资助:
    国家重点基础研究发展计划;国家973项目

Regulation of inflammatory cytokine gene transcription in macrophage induced by P.yoelii MIF

Ding-ding SHAO1,Shan ZENG2,Xiang ZHONG2,Heng WANG1   

  1. 1. School of Basic Medicine IBMS, CAMS & PUMC
    2.
  • Received:2011-03-28 Revised:2011-05-10 Online:2011-07-05 Published:2011-07-05
  • Contact: Heng WANG E-mail:wanghpumc@gmail.com

摘要: 目的 分析约氏疟原虫表达的巨噬细胞迁移抑制因子(MIF) 对小鼠巨噬细胞炎性因子基因表达的调节情况,并比较PyMIF和宿主小鼠MIF (MmMIF) 在调节炎性因子表达水平方面的差异。方法 亲和柱纯化原核表达的带His 标签的重组MIF 蛋白, 去除内毒素后刺激体外培养的同步化小鼠单核巨噬细胞系, 随后纯化高质量的RNA , 通过小鼠炎性因子和受体芯片进行分析。结果 PyMIF显著促进(ΔΔCt≥2)了17个炎性因子和炎性因子受体基因的转录,同时显著降低(ΔΔCt≤ -2)了3个基因(CCR6、CCR8和IL1F6)的转录水平。并且,PyMIF调节的基因数量明显多于MmMIF。结论 PyMIF和MmMIF对巨噬细胞的活性调节存在显著差异,此结果为进一步了解PyMIF对宿主炎性反应的作用提供了重要数据。

关键词: 巨噬细胞迁移抑制因子, 疟原虫, 基因芯片, 炎症因子

Abstract: Objective Plasmodium derived macrophage migration inhibitory factor (MIF) ortholog was considered to have potential to regulate host immune cell. Plasmodium yoelii derived MIF (PyMIF) has highly conserved crystal structure and similar bioactivity with host mouse MIF. However, the regulatory mechanism, including the function of PyMIF on host macrophage, is not clear so far. Here, we report the regulation of PyMIF on host inflammatory cytokines transcription in macrophage, and the compared analysis of regulation activity between PyMIF and host mouse MIF (MmMIF). Methods Both PyMIF and MmMIF were expressed and purified as recombinant proteins and were removed the endotoxin by C8 reversed-phase column. Then the endotoxin-free protein was added to cultured mouse monocyte / macrophage cell line RAW264.7. After incubation, the total RNA was extracted and applied to "RT2 Profiler? PCR Array Mouse Inflammatory Cytokines & Receptors" analysis. Results PyMIF significantly up-regulated 17 inflammatory cytokine genes transcription level and down-regulated 3 genes (CCR6、CCR8 and IL1F6) transcription. The regulated gene quantity by PyMIF was more than MmMIF. Most genes regulated by PyMIF were involved in a Th2 response bias or in expanding inflammatory response. Conclusions There is much difference between PyMIF and MmMIF in regulating host macrophage activity. These results provided more information about Plasmodium MIF function on host immune cell, and will be meaning for deep understanding the role of Plasmodium MIF during infection.

Key words: macrophage migration inhibitory factor, Plasmodium, bio-chip, inflammatory cytokine

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