Quantitative isolation and in vivo imaging of malaria parasite liver stages
Quantitative isolation and in vivo imaging of malaria parasite liver stages
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DOI：
10.1016/j.ijpara.2006.06.009
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发表时间：
2006-10-01
影响因子：
4
通讯作者：
Kappe, Stefan H. I.
中科院分区：
文献类型：
--
作者：
Tarun, Alice S.;Baer, Kerstin;Kappe, Stefan H. I.
疟疾的病原体疟原虫的肝脏阶段是寄生虫生命周期中探索最少的形式，尽管它们被认为是关键的疫苗和药物靶点。人体疟疾寄生虫肝脏阶段的体内实验实际上是被禁止的，因此啮齿动物模型疟疾寄生虫已被用于体内研究。然而，即使在啮齿动物模型中，肝脏分期分析的进展也受到限制，主要是由于它们的丰度低以及相关的可视化和分离困难。在这里，我们提出了绿色荧光蛋白（GFP）标记的BALB/c小鼠的约利疟原虫啮齿动物疟疾寄生虫肝脏感染作为一种极好的定量模型，用于活体可视化和分离迄今为止难以捉摸的肝脏阶段。我们相信P. yoelii gfp标记的肝分期首次允许通过荧光激活细胞分选有效地定量分离完整的早期和晚期肝感染的肝细胞单位。gfp标记的肝脏分期也非常适合活体成像，使我们第一次能够实时可视化它们。我们确定了以前未被认识到的肝脏阶段的特征，包括剧烈的寄生虫运动和“挤出体”的排出。因此，活体成像揭示了疟疾寄生虫从组织阶段过渡到血液阶段的新的重要信息。(c) 2006年澳大利亚寄生虫学学会Elsevier Ltd.出版。版权所有。
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. (c) 2006 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved.