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疟原虫肝脏期的定量分离与体内成像

Quantitative isolation and in vivo imaging of malaria parasite liver stages.

作者信息

Tarun Alice S, Baer Kerstin, Dumpit Ronald F, Gray Sean, Lejarcegui Nicholas, Frevert Ute, Kappe Stefan H I

机构信息

Seattle Biomedical Research Institute, Seattle, WA 98109, USA.

出版信息

Int J Parasitol. 2006 Oct;36(12):1283-93. doi: 10.1016/j.ijpara.2006.06.009. Epub 2006 Jul 12.

Abstract

The liver stages of Plasmodium, the causative agent of malaria, are the least explored forms in the parasite's life cycle despite their recognition as key vaccine and drug targets. In vivo experimental access to liver stages of human malaria parasites is practically prohibited and therefore rodent model malaria parasites have been used for in vivo studies. However, even in rodent models progress in the analysis of liver stages has been limited, mainly due to their low abundance and associated difficulties in visualisation and isolation. Here, we present green fluorescent protein (GFP)-tagged Plasmodium yoelii rodent malaria parasite liver infections in BALB/c mice as an excellent quantitative model for the live visualisation and isolation of the so far elusive liver stages. We believe P. yoelii GFP-tagged liver stages allow, for the first time, the efficient quantitative isolation of intact early and late liver stage-infected hepatocyte units by fluorescence activated cell sorting. GFP-tagged liver stages are also well suited for intravital imaging, allowing us for the first time to visualise them in real time. We identify previously unrecognised features of liver stages including vigorous parasite movement and expulsion of 'extrusomes'. Intravital imaging thus reveals new, important information on the malaria parasite's transition from tissue to blood stage.

摘要

疟原虫是疟疾的病原体,尽管其肝脏阶段被认为是关键的疫苗和药物靶点,但却是该寄生虫生命周期中研究最少的形式。实际上,禁止在人体上对疟原虫肝脏阶段进行体内实验,因此啮齿动物模型疟原虫已被用于体内研究。然而,即使在啮齿动物模型中,肝脏阶段分析的进展也很有限,主要是因为其丰度低以及在可视化和分离方面存在相关困难。在此,我们展示了在BALB/c小鼠中感染绿色荧光蛋白(GFP)标记的约氏疟原虫啮齿动物疟疾寄生虫肝脏,这是一种用于实时可视化和分离迄今为止难以捉摸的肝脏阶段的优秀定量模型。我们相信,带有GFP标记的约氏疟原虫肝脏阶段首次实现了通过荧光激活细胞分选对完整的早期和晚期肝脏阶段感染的肝细胞单元进行高效定量分离。带有GFP标记的肝脏阶段也非常适合活体成像,使我们首次能够实时观察它们。我们发现了肝脏阶段以前未被认识到的特征,包括寄生虫的剧烈运动和“挤出体”的排出。活体成像因此揭示了有关疟原虫从组织阶段向血液阶段转变的新的重要信息。

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