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Transgenic Fluorescent Plasmodium cynomolgi Liver Stages Enable Live Imaging and Purification of Malaria Hypnozoite-Forms

A major challenge for strategies to combat the human malaria parasite Plasmodium vivax is the presence of hypnozoites in the liver. These dormant forms can cause renewed clinical disease after reactivation through unknown mechanisms. The closely related non-human primate malaria P. cynomolgi is a fr...

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Autores principales: Voorberg-van der Wel, Annemarie, Zeeman, Anne-Marie, van Amsterdam, Sandra M., van den Berg, Alexander, Klooster, Els J., Iwanaga, Shiroh, Janse, Chris J., van Gemert, Geert-Jan, Sauerwein, Robert, Beenhakker, Niels, Koopman, Gerrit, Thomas, Alan W., Kocken, Clemens H. M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3554669/
https://www.ncbi.nlm.nih.gov/pubmed/23359816
http://dx.doi.org/10.1371/journal.pone.0054888
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author Voorberg-van der Wel, Annemarie
Zeeman, Anne-Marie
van Amsterdam, Sandra M.
van den Berg, Alexander
Klooster, Els J.
Iwanaga, Shiroh
Janse, Chris J.
van Gemert, Geert-Jan
Sauerwein, Robert
Beenhakker, Niels
Koopman, Gerrit
Thomas, Alan W.
Kocken, Clemens H. M.
author_facet Voorberg-van der Wel, Annemarie
Zeeman, Anne-Marie
van Amsterdam, Sandra M.
van den Berg, Alexander
Klooster, Els J.
Iwanaga, Shiroh
Janse, Chris J.
van Gemert, Geert-Jan
Sauerwein, Robert
Beenhakker, Niels
Koopman, Gerrit
Thomas, Alan W.
Kocken, Clemens H. M.
author_sort Voorberg-van der Wel, Annemarie
collection PubMed
description A major challenge for strategies to combat the human malaria parasite Plasmodium vivax is the presence of hypnozoites in the liver. These dormant forms can cause renewed clinical disease after reactivation through unknown mechanisms. The closely related non-human primate malaria P. cynomolgi is a frequently used model for studying hypnozoite-induced relapses. Here we report the generation of the first transgenic P. cynomolgi parasites that stably express fluorescent markers in liver stages by transfection with novel DNA-constructs containing a P. cynomolgi centromere. Analysis of fluorescent liver stages in culture identified, in addition to developing liver-schizonts, uninucleate persisting parasites that were atovaquone resistant but primaquine sensitive, features associated with hypnozoites. We demonstrate that these hypnozoite-forms could be isolated by fluorescence-activated cell sorting. The fluorescently-tagged parasites in combination with FACS-purification open new avenues for a wide range of studies for analysing hypnozoite biology and reactivation.
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spelling pubmed-35546692013-01-28 Transgenic Fluorescent Plasmodium cynomolgi Liver Stages Enable Live Imaging and Purification of Malaria Hypnozoite-Forms Voorberg-van der Wel, Annemarie Zeeman, Anne-Marie van Amsterdam, Sandra M. van den Berg, Alexander Klooster, Els J. Iwanaga, Shiroh Janse, Chris J. van Gemert, Geert-Jan Sauerwein, Robert Beenhakker, Niels Koopman, Gerrit Thomas, Alan W. Kocken, Clemens H. M. PLoS One Research Article A major challenge for strategies to combat the human malaria parasite Plasmodium vivax is the presence of hypnozoites in the liver. These dormant forms can cause renewed clinical disease after reactivation through unknown mechanisms. The closely related non-human primate malaria P. cynomolgi is a frequently used model for studying hypnozoite-induced relapses. Here we report the generation of the first transgenic P. cynomolgi parasites that stably express fluorescent markers in liver stages by transfection with novel DNA-constructs containing a P. cynomolgi centromere. Analysis of fluorescent liver stages in culture identified, in addition to developing liver-schizonts, uninucleate persisting parasites that were atovaquone resistant but primaquine sensitive, features associated with hypnozoites. We demonstrate that these hypnozoite-forms could be isolated by fluorescence-activated cell sorting. The fluorescently-tagged parasites in combination with FACS-purification open new avenues for a wide range of studies for analysing hypnozoite biology and reactivation. Public Library of Science 2013-01-24 /pmc/articles/PMC3554669/ /pubmed/23359816 http://dx.doi.org/10.1371/journal.pone.0054888 Text en © 2013 Voorberg-van der Wel et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Voorberg-van der Wel, Annemarie
Zeeman, Anne-Marie
van Amsterdam, Sandra M.
van den Berg, Alexander
Klooster, Els J.
Iwanaga, Shiroh
Janse, Chris J.
van Gemert, Geert-Jan
Sauerwein, Robert
Beenhakker, Niels
Koopman, Gerrit
Thomas, Alan W.
Kocken, Clemens H. M.
Transgenic Fluorescent Plasmodium cynomolgi Liver Stages Enable Live Imaging and Purification of Malaria Hypnozoite-Forms
title Transgenic Fluorescent Plasmodium cynomolgi Liver Stages Enable Live Imaging and Purification of Malaria Hypnozoite-Forms
title_full Transgenic Fluorescent Plasmodium cynomolgi Liver Stages Enable Live Imaging and Purification of Malaria Hypnozoite-Forms
title_fullStr Transgenic Fluorescent Plasmodium cynomolgi Liver Stages Enable Live Imaging and Purification of Malaria Hypnozoite-Forms
title_full_unstemmed Transgenic Fluorescent Plasmodium cynomolgi Liver Stages Enable Live Imaging and Purification of Malaria Hypnozoite-Forms
title_short Transgenic Fluorescent Plasmodium cynomolgi Liver Stages Enable Live Imaging and Purification of Malaria Hypnozoite-Forms
title_sort transgenic fluorescent plasmodium cynomolgi liver stages enable live imaging and purification of malaria hypnozoite-forms
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3554669/
https://www.ncbi.nlm.nih.gov/pubmed/23359816
http://dx.doi.org/10.1371/journal.pone.0054888
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