疟原虫源巨噬细胞迁移抑制因子的分泌途径及其对宿主细胞基因表达影响的初步研究

发布时间：2018-01-12 05:16

本文关键词：疟原虫源巨噬细胞迁移抑制因子的分泌途径及其对宿主细胞基因表达影响的初步研究　出处：《中国协和医科大学》2008年硕士论文　论文类型：学位论文

【摘要】： 疟疾是世界上最严重的传染性疾病之一,由于抗药虫株的出现以及尚无有效的疫苗,目前疟疾的防治仍然面临巨大挑战。迄今为止,疟疾病理机制、疟原虫逃逸宿主免疫攻击机制等方面仍有大量疑问尚未解决,这主要是由于目前人们对疟原虫生物学的认识还十分有限所致。本实验室前期分别鉴定了两种来源于疟原虫的巨噬细胞迁移抑制因子(macrophage migration inhibitory factor,MIF):恶性疟原虫PfMIF(Plasmodium falciparum MIF)和约氏疟原虫PyMIF(Plasmodiumyoelii MIF),并且证明疟原虫MIF分子具有MIF家族的典型结构特征和生物学活性,并建立了特异检测技术。本论文工作在此基础上,对疟原虫MIF分子的分泌途径和其参与的信号通路以及诱导的免疫效应等方面进行了初步探讨。实验结果显示,在转染的哺乳动物细胞系中PfMIF分子的分泌仅被抑制剂Glyburide显著抑制,提示PfMIF的分泌通过非经典分泌途径ABC转运家族成员ABCA1实现,与高等动物MIF分子的分泌途径一致。在体外培养的恶性疟原虫感染红细胞中,Methylamine显著抑制PfMIF分泌,而Glyburide没有明显效应,提示在感染红细胞中PfMIF的分泌可能是通过内吞小泡系统实现。另外,应用三种以real-time PCR技术为基础的第二代芯片,证实PyMIF和宿主小鼠MuMIF能够调节多种免疫效应分子和信号通路分子的转录,其中PyMIF对多种趋化因子及其受体基因、癌基因、凋亡相关基因等均有显著的调节效应。同时为了验证芯片实验的结果,本论文工作同时将部分基因进行半定量RT-PCR验证,结果与芯片实验一致。本论文工作初步揭示了疟原虫来源MIF分子的分泌途径,并证实其能够调节宿主细胞多种炎症相关基因和信号通路基因的转录,有助于进一步理解疟原虫生物学本质,并为下一步继续揭示疟原虫MIF分子在感染过程中的作用提供重要提示。
[Abstract]:Malaria is one of the most serious infectious diseases in the world. Due to the emergence of drug-resistant strains and the absence of effective vaccines, the control of malaria is still facing great challenges. There are still a lot of questions about the immune attack mechanism of malaria parasite escape host. This is mainly due to the limited understanding of the biology of Plasmodium. Two kinds of macrophage migration inhibitory factors from Plasmodium have been identified in our laboratory. Macrophage migration inhibitory factor. PfMIF(Plasmodium falciparum MIF and Plasmodium yoelii PyMIF. Plasmodiumyoelii MIF. It is proved that the MIF molecule of Plasmodium falciparum has the typical structural characteristics and biological activity of MIF family, and a specific detection technique has been established. The secretory pathway and signal pathway of Plasmodium MIF molecule and its induced immune effect were discussed in this paper. The results showed that the secretion of PfMIF molecules in transfected mammalian cell lines was significantly inhibited only by the inhibitor Glyburide. The results suggest that the secretion of PfMIF is realized by ABCA1 of ABC transporter family through non-classical secretory pathway. In accordance with the secretory pathway of MIF molecules in higher animals, Methylamine significantly inhibited the secretion of PfMIF in erythrocytes infected with Plasmodium falciparum in vitro. However, Glyburide has no obvious effect, suggesting that the secretion of PfMIF in infected red blood cells may be realized by endocytosis vesicle system. Using three kinds of second generation chips based on real-time PCR technology, it was proved that PyMIF and host mouse MuMIF could regulate the transcription of various immune effector molecules and signal pathway molecules. PyMIF has a significant regulatory effect on many chemokines and their receptor genes, oncogenes, apoptosis-related genes and so on. At the same time, some genes were verified by semi-quantitative RT-PCR, and the results were consistent with the chip experiments. In this paper, the secretory pathway of MIF derived from Plasmodium has been preliminarily revealed, and it has been proved that it can regulate the transcription of many inflammatory genes and signaling pathway genes in host cells. It is helpful to further understand the biological nature of Plasmodium and provide important clues for the next step to reveal the role of Plasmodium MIF molecules in the process of infection.
【学位授予单位】：中国协和医科大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R382

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