title: Persistence and activation of malaria hypnozoites in long-term primary hepatocyte cultures.
abstract: Malaria relapses, resulting from the activation of quiescent hepatic hypnozoites of Plasmodium vivax and Plasmodium ovale, hinder global efforts to control and eliminate malaria. As primaquine, the only drug capable of eliminating hypnozoites, is unsuitable for mass administration, an alternative drug is needed urgently. Currently, analyses of hypnozoites, including screening of compounds that would eliminate them, can only be made using common macaque models, principally Macaca rhesus and Macaca fascicularis, experimentally infected with the relapsing Plasmodium cynomolgi. Here, we present a protocol for long-term in vitro cultivation of P. cynomolgi-infected M. fascicularis primary hepatocytes during which hypnozoites persist and activate to resume normal development. In a proof-of-concept experiment, we obtained evidence that exposure to an inhibitor of histone modification enzymes implicated in epigenetic control of gene expression induces an accelerated rate of hypnozoite activation. The protocol presented may further enable investigations of hypnozoite biology and the search for compounds that kill hypnozoites or disrupt their quiescence.
authors: 1] Sorbonne Universités, Université Pierre et Marie Curie Paris 6, UMR S945, Paris, France. [2] Institut National de la Santé et de la Recherche Médicale, U945, Paris, France. [3].
authors: Laurent
authors: L
authors: Dembélé
authors: 1] Sorbonne Universités, Université Pierre et Marie Curie Paris 6, UMR S945, Paris, France. [2] Institut National de la Santé et de la Recherche Médicale, U945, Paris, France. [3].
authors: Jean-François
authors: JF
authors: Franetich
authors: 1] Sorbonne Universités, Université Pierre et Marie Curie Paris 6, UMR S945, Paris, France. [2] Institut National de la Santé et de la Recherche Médicale, U945, Paris, France.
authors: Audrey
authors: A
authors: Lorthiois
authors: 1] Sorbonne Universités, Université Pierre et Marie Curie Paris 6, UMR S945, Paris, France. [2] Institut National de la Santé et de la Recherche Médicale, U945, Paris, France.
authors: Audrey
authors: A
authors: Gego
authors: Department of Parasitology, Biomedical Primate Research Centre, Rijswijk, The Netherlands.
authors: Anne-Marie
authors: AM
authors: Zeeman
authors: Department of Parasitology, Biomedical Primate Research Centre, Rijswijk, The Netherlands.
authors: Clemens H M
authors: CH
authors: Kocken
authors: 1] Division of Immuno-Virology, Institute of Emerging Diseases and Innovative Therapies, Commissariat à l'Energie Atomique et aux Energies Alternatives, Fontenay-aux-Roses, France. [2] Université Paris-Sud XI, UMRE01, Orsay, France.
authors: Roger
authors: R
authors: Le Grand
authors: 1] Division of Immuno-Virology, Institute of Emerging Diseases and Innovative Therapies, Commissariat à l'Energie Atomique et aux Energies Alternatives, Fontenay-aux-Roses, France. [2] Université Paris-Sud XI, UMRE01, Orsay, France.
authors: Nathalie
authors: N
authors: Dereuddre-Bosquet
authors: Department of Medical Microbiology, Radboud University Nijmegen Medical Centre, Nijmegen, The Netherlands.
authors: Geert-Jan
authors: GJ
authors: van Gemert
authors: Department of Medical Microbiology, Radboud University Nijmegen Medical Centre, Nijmegen, The Netherlands.
authors: Robert
authors: R
authors: Sauerwein
authors: Service de Chirurgie Digestive, Hépato-Bilio-Pancréatique et Transplantation Hépatique, Hôpital Pitié-Salpêtrière, Paris, France.
authors: Jean-Christophe
authors: JC
authors: Vaillant
authors: Service de Chirurgie Digestive, Hépato-Bilio-Pancréatique et Transplantation Hépatique, Hôpital Pitié-Salpêtrière, Paris, France.
authors: Laurent
authors: L
authors: Hannoun
authors: Department of Chemistry, Imperial College London, South Kensington Campus, London, UK.
authors: Matthew J
authors: MJ
authors: Fuchter
authors: Novartis Institute for Tropical Diseases, Singapore.
authors: Thierry T
authors: TT
authors: Diagana
authors: 1] Unité de Biologie des Interactions Hôte-Parasite, Institut Pasteur, Paris, France. [2] Centre National de la Recherche Scientifique, Unité de Recherche Associée 2581, Paris, France.
authors: Nicholas A
authors: NA
authors: Malmquist
authors: 1] Unité de Biologie des Interactions Hôte-Parasite, Institut Pasteur, Paris, France. [2] Centre National de la Recherche Scientifique, Unité de Recherche Associée 2581, Paris, France.
authors: Artur
authors: A
authors: Scherf
authors: 1] Sorbonne Universités, Université Pierre et Marie Curie Paris 6, UMR S945, Paris, France. [2] Institut National de la Santé et de la Recherche Médicale, U945, Paris, France.
authors: Georges
authors: G
authors: Snounou
authors: 1] Sorbonne Universités, Université Pierre et Marie Curie Paris 6, UMR S945, Paris, France. [2] Institut National de la Santé et de la Recherche Médicale, U945, Paris, France. [3] Assistance Publique-Hôpitaux de Paris, Groupe Hospitalier Pitié-Salpêtrière, Service Parasitologie-Mycologie, Paris, France.
authors: Dominique
authors: D
authors: Mazier
mesh_terms: D000818:Animals; D002460:Cell Line; D002478:Cells, Cultured; D044127:Epigenesis, Genetic; D005260:Female; D049452:Green Fluorescent Proteins / Q000737:chemistry; D022781:Hepatocytes / Q000469:parasitology*; D000076983:Histone Methyltransferases; D011495:Histone-Lysine N-Methyltransferase / Q000037:antagonists & inhibitors; D006657:Histones / Q000737:chemistry; D006801:Humans; D008251:Macaca / Q000469:parasitology; D008288:Malaria / Q000469:parasitology*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008856:Microscopy, Fluorescence; D016789:Plasmodium cynomolgi / Q000254:growth & development*; D012008:Recurrence; D034101:Sporozoites / Q000254:growth & development*; D013997:Time Factors
chemical_list: D006657:Histones; D049452:Green Fluorescent Proteins; D000076983:Histone Methyltransferases; D011495:Histone-Lysine N-Methyltransferase