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A dual fluorescent Plasmodium cynomolgi reporter line reveals in vitro malaria hypnozoite reactivation.
Communications Biology ( IF 5.2 ) Pub Date : 2020-01-03 , DOI: 10.1038/s42003-019-0737-3
Annemarie M Voorberg-van der Wel 1 , Anne-Marie Zeeman 1 , Ivonne G Nieuwenhuis 1 , Nicole M van der Werff 1 , Els J Klooster 1 , Onny Klop 1 , Lars C Vermaat 1 , Devendra Kumar Gupta 2 , Laurent Dembele 2, 3 , Thierry T Diagana 2 , Clemens H M Kocken 1
Affiliation  

Plasmodium vivax malaria is characterized by repeated episodes of blood stage infection (relapses) resulting from activation of dormant stages in the liver, so-called hypnozoites. Transition of hypnozoites into developing schizonts has never been observed. A barrier for studying this has been the lack of a system in which to monitor growth of liver stages. Here, exploiting the unique strengths of the simian hypnozoite model P. cynomolgi, we have developed green-fluorescent (GFP) hypnozoites that turn on red-fluorescent (mCherry) upon activation. The transgenic parasites show full liver stage development, including merozoite release and red blood cell infection. We demonstrate that individual hypnozoites actually can activate and resume development after prolonged culture, providing the last missing evidence of the hypnozoite theory of relapse. The few events identified indicate that hypnozoite activation in vitro is infrequent. This system will further our understanding of the mechanisms of hypnozoite activation and may facilitate drug discovery approaches.

中文翻译:

双重荧光的食蟹疟原虫报告基因系显示了体外疟疾次生子再活化。

间日疟原虫疟疾的特征是反复发作的血液阶段感染(复发)是由于肝脏休眠阶段的激活引起的,即所谓的次生动物。从未观察到次生孢子向发育中的裂殖体的过渡。研究这一问题的障碍是缺乏监测肝阶段生长的系统。在这里,我们利用猿猴次生动物模型食蟹猴P.cynomolgi的独特优势,开发了在激活时会打开红色荧光(mCherry)的绿色荧光(GFP)次生动物。转基因寄生虫显示出完整的肝发育阶段,包括裂殖子释放和红细胞感染。我们证明,经过长时间的培养,各个次生动物实际上可以激活并恢复发育,为次生动物的复发理论提供了最后的证据。鉴定出的少数事件表明,次生子体外活化很少。该系统将进一步加深我们对次裂殖子活化机制的理解,并可能有助于药物发现方法。
更新日期:2020-01-04
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