Activity of clinically relevant antimalarial drugs on Plasmodium falciparum mature gametocytes in an ATP bioluminescence "transmission blocking" assay.
Activity of clinically relevant antimalarial drugs on Plasmodium falciparum mature gametocytes in an ATP bioluminescence "transmission blocking" assay.
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DOI：
10.1371/journal.pone.0035019
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Herreros E
中科院分区：
文献类型：
--
作者：
Lelièvre J;Almela MJ;Lozano S;Miguel C;Franco V;Leroy D;Herreros E
目前的抗疟疾药物是根据它们对引起症状的寄生虫无性血液阶段的活性来选择的。这些药物中的哪一种也针对配子体，在性阶段负责疾病传播，仍然未知。阻断传播是根除议程中的主要策略之一，需要鉴定对配子体有活性的新分子。然而，迄今为止，大规模测量分子对成熟配子体的影响的主要限制是缺乏标准化和可靠的方法。本研究提供了一种体外制备和纯化成熟配子体的有效方法。基于这种新方法，我们开发了一种强大、经济、灵敏的ATP生物发光检测方法。然后，我们评估了17种金标准抗疟疾药物对疟原虫晚期配子体的活性。生产大量配子体的困难限制了疟疾传播阻断试验的发展进展。我们改进了Ifediba和Vanderberg建立的方法，无论使用哪种菌株，都能获得大量的成熟配子体。我们设计了一种测定抗疟药物活性的方法，该方法基于纯化的IV-V期配子体在96/384孔微孔板上暴露48小时后细胞内ATP含量。我们还测量了药物在无性期的活性和对HepG2细胞的细胞毒性，以估计活性药物的特异性。这里所描述的工作代表了用适合中/高通量筛选的自动化试验确定任何菌株成熟配子体上新分子活性的又一重要步骤。考虑到有性和无性阶段所涉及的形式的生物学是非常不同的，我们的200万种化合物库的筛选可能使我们能够发现针对配子细胞特异性代谢途径的新型抗疟疾药物。
Current anti-malarial drugs have been selected on the basis of their activity against the symptom-causing asexual blood stage of the parasite. Which of these drugs also target gametocytes, in the sexual stage responsible for disease transmission, remains unknown. Blocking transmission is one of the main strategies in the eradication agenda and requires the identification of new molecules that are active against gametocytes. However, to date, the main limitation for measuring the effect of molecules against mature gametocytes on a large scale is the lack of a standardized and reliable method. Here we provide an efficient method to produce and purify mature gametocytes in vitro. Based on this new procedure, we developed a robust, affordable, and sensitive ATP bioluminescence-based assay. We then assessed the activity of 17 gold-standard anti-malarial drugs on Plasmodium late stage gametocytes. Difficulties in producing large amounts of gametocytes have limited progress in the development of malaria transmission blocking assays. We improved the method established by Ifediba and Vanderberg to obtain viable, mature gametocytes en masse, whatever the strain used. We designed an assay to determine the activity of antimalarial drugs based on the intracellular ATP content of purified stage IV–V gametocytes after 48 h of drug exposure in 96/384-well microplates. Measurements of drug activity on asexual stages and cytotoxicity on HepG2 cells were also obtained to estimate the specificity of the active drugs. The work described here represents another significant step towards determination of the activity of new molecules on mature gametocytes of any strain with an automated assay suitable for medium/high-throughput screening. Considering that the biology of the forms involved in the sexual and asexual stages is very different, a screen of our 2 million-compound library may allow us to discover novel anti-malarial drugs to target gametocyte-specific metabolic pathways.
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影响因子：
3.7
作者：
Coulibaly B;Zoungrana A;Mockenhaupt FP;Schirmer RH;Klose C;Mansmann U;Meissner PE;Müller O
通讯作者：
Müller O
影响因子：
2
作者：
LANNERS, HN
通讯作者：
LANNERS, HN
影响因子：
4.9
作者：
Benoit-Vical, Francoise;Lelievre, Joel;Meunier, Bernard
通讯作者：
Meunier, Bernard
DOI：
10.1111/j.1550-7408.1985.tb03020.x
发表时间：
1985-01-01
期刊：
JOURNAL OF PROTOZOOLOGY
影响因子：
--
作者：
GUPTA, SK;SCHULMAN, S;VANDERBERG, JP
通讯作者：
VANDERBERG, JP
影响因子：
2.2
作者：
CROUCH, SPM;KOZLOWSKI, R;FLETCHER, J
通讯作者：
FLETCHER, J