Visualisation and quantitative analysis of the rodent malaria liver stage by real time imaging

Ploemen, I.H.J. et al. (2010) Visualisation and quantitative analysis of the rodent malaria liver stage by real time imaging. PLoS ONE, 4(11), e7881. (doi:10.1371/journal.pone.0007881)

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Abstract

The quantitative analysis of Plasmodium development in the liver in laboratory animals in cultured cells is hampered by low parasite infection rates and the complicated methods required to monitor intracellular development. As a consequence, this important phase of the parasite's life cycle has been poorly studied compared to blood stages, for example in screening anti-malarial drugs. Here we report the use of a transgenic P. berghei parasite, Pb GFP-Luc con, expressing the bioluminescent reporter protein luciferase to visualize and quantify parasite development in liver cells both in culture and in live mice using real-time luminescence imaging. The reporter-parasite based quantification in cultured hepatocytes by real-time imaging or using a microplate reader correlates very well with established quantitative RT-PCR methods. For the first time the liver stage of <italic>Plasmodium</italic> is visualized in whole bodies of live mice and we were able to discriminate as few as 1GÇô5 infected hepatocytes per liver in mice using 2D-imaging and to identify individual infected hepatocytes by 3D-imaging. The analysis of liver infections by whole body imaging shows a good correlation with quantitative RT-PCR analysis of extracted livers. The luminescence-based analysis of the effects of various drugs on in vitro hepatocyte infection shows that this method can effectively be used for <italic>in vitro</italic> screening of compounds targeting Plasmodium liver stages. Furthermore, by analysing the effect of primaquine and tafenoquine in vivo we demonstrate the applicability of real time imaging to assess parasite drug sensitivity in the liver. The simplicity and speed of quantitative analysis of liver-stage development by real-time imaging compared to the PCR methodologies, as well as the possibility to analyse liver development in live mice without surgery, opens up new possibilities for research on Plasmodium liver infections and for validating the effect of drugs and vaccines on the liver stage of Plasmodium.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Waters, Professor Andy
Authors: Ploemen, I.H.J., Prudêncio, M., Douradinha, B.G., Ramesar, J., Fonager, J., van Gemert, G.J., Luty, A.J.F., Hermsen, C.C., Sauerwein, R.W., Baptista, F.G., Mota, M.M., Waters, A.P., Que, I., Lowik, C.W.G.M., Khan, S.M., Janse, C.J., and Franke-Fayard, B.M.D.
College/School:College of Medical Veterinary and Life Sciences > Institute of Infection Immunity and Inflammation
Journal Name:PLoS ONE
ISSN:1932-6203
ISSN (Online):1932-6203
Copyright Holders:© 2009 Ploemen et al.
First Published:First published in PLoS ONE 4(11): e7881.
Publisher Policy:This is an open-access article distributed under the terms of the Creative Commons Attribution License

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