Dynasore, a dynamin inhibitor, inhibits Trypanosoma cruzi entry into peritoneal macrophages.
Dynasore, a dynamin inhibitor, inhibits Trypanosoma cruzi entry into peritoneal macrophages.
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Dynasore是一种动力蛋白抑制剂，可抑制克鲁齐锥虫进入腹膜巨噬细胞。
DOI：
10.1371/journal.pone.0007764
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发表时间：
2010-01-20
期刊：
影响因子：
3.7
通讯作者：
Carvalho TM
中科院分区：
文献类型：
--
作者：
Barrias ES;Reignault LC;De Souza W;Carvalho TM
克氏锥虫是一种细胞内寄生虫，像其他一些细胞内病原体一样，靶向宿主细胞囊泡运输机制的特定蛋白质，导致宿主细胞过程的调节，从而产生独特的吞噬体。在哺乳动物细胞中，已经确定了几种分子选择性地调节内吞运输囊泡的形成以及这些囊泡与适当的受体膜的融合。其中，GTPase动力蛋白在网格蛋白介导的胞吞作用中起重要作用，最近发现动力蛋白可参与吞噬过程。我们使用了一种叫做dynamore的化合物，它能够阻断dynamin的GTPase活性。Dynasore作为一种有效的内吞途径抑制剂，通过阻断其添加后几秒钟内的包被囊泡形成。本研究利用动态蛋白（dynamin）对克氏锥虫进入吞噬细胞和非吞噬细胞的过程进行了研究。在此目的中，腹腔巨噬细胞和LLC-MK2细胞在与色乳突、无乳突或粘乳突相互作用之前，用增加浓度的dynasore处理。我们观察到，在这两种细胞系中，当我们使用100µM dynasore时，寄生虫内化急剧减少（lc - mk2细胞减少90%以上，腹膜巨噬细胞减少70%）。然而，两种细胞系的克氏体黏附指数均未受影响。通过扫描电镜分析这些相互作用，并将腹腔巨噬细胞与LLC-MK2细胞进行比较，发现细胞进入被阻断的阶段存在差异。在lc - mk2细胞中，这种阻滞比在腹膜巨噬细胞中更早被观察到。在lc - mk2细胞中，寄生虫仅与细胞微绒毛相关，而在腹膜巨噬细胞中，锥乳线虫并未完全被宿主细胞膜吞噬。综上所述，我们的研究结果表明，动力蛋白是宿主细胞在与克氏锥虫相互作用过程中侵袭细胞，特别是寄生液泡形成所必需的分子。
Trypanosoma cruzi is an intracellular parasite that, like some other intracellular pathogens, targets specific proteins of the host cell vesicular transport machinery, leading to a modulation of host cell processes that results in the generation of unique phagosomes. In mammalian cells, several molecules have been identified that selectively regulate the formation of endocytic transport vesicles and the fusion of such vesicles with appropriate acceptor membranes. Among these, the GTPase dynamin plays an important role in clathrin-mediated endocytosis, and it was recently found that dynamin can participate in a phagocytic process. We used a compound called dynasore that has the ability to block the GTPase activity of dynamin. Dynasore acts as a potent inhibitor of endocytic pathways by blocking coated vesicle formation within seconds of its addition. Here, we investigated whether dynamin is involved in the entry process of T. cruzi in phagocytic and non-phagocytic cells by using dynasore. In this aim, peritoneal macrophages and LLC-MK2 cells were treated with increasing concentrations of dynasore before interaction with trypomastigotes, amastigotes or epimastigotes. We observed that, in both cell lines, the parasite internalization was drastically diminished (by greater than 90% in LLC-MK2 cells and 70% in peritoneal macrophages) when we used 100 µM dynasore. The T. cruzi adhesion index, however, was unaffected in either cell line. Analyzing these interactions by scanning electron microscopy and comparing peritoneal macrophages to LLC-MK2 cells revealed differences in the stage at which cell entry was blocked. In LLC-MK2 cells, this blockade is observed earlier than it is in peritoneal macrophages. In LLC-MK2 cells, the parasites were only associated with cellular microvilli, whereas in peritoneal macrophages, trypomastigotes were not completely engulfed by a host cell plasma membrane. Taken together our results demonstrate that dynamin is an essential molecule necessary for cell invasion and specifically parasitophorous vacuole formation by host cells during interaction with Trypanosoma cruzi.
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影响因子：
15.3
作者：
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