Temperature shift and host cell contact up-regulate sporozoite expression of Plasmodium falciparum genes involved in hepatocyte infection
| Autor: | Geert-Jaan van Gemert, Emmanuel Bischoff, Samir Yalaoui, Dominique Mazier, Patrick Froissard, Carine Marinach, Jean-François Franetich, Georges Snounou, Catherine Vaquero, Olivier Silvie, Anthony Siau, Peter H. David, Laurent Hannoun, Adrian J. F. Luty |
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| Přispěvatelé: | Immunobiologie Cellulaire et Moléculaire des Infections Parasitaires, Université Pierre et Marie Curie - Paris 6 (UPMC)-IFR113-Institut National de la Santé et de la Recherche Médicale (INSERM), Service de Chirurgie Digestive, Hépato-Bilio-pancréatique et Transplantation Hépatique [CHU Pitié-Salpétrière], CHU Pitié-Salpêtrière [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), Department of Medical Microbiology [Nijmegen], Radboud University Medical Center [Nijmegen], Génopole, Institut Pasteur [Paris] (IP), Parasitologie moléculaire et Signalisation, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Parasitologie Comparée et Modèles Expérimentaux, Muséum national d'Histoire naturelle (MNHN)-Centre National de la Recherche Scientifique (CNRS), Service de Parasitologie - Mycologie [CHU Pitié-Salpétrière], Plasmodium pan genomic microarrays were financed by grants from the Délégation Générale pour l’Armement (DGA verst.nu22120/DSP/SREAF and Contract no 0434025), Programme PAL+/Fond National pour la Science, the Institut Pasteur and the Programme Génopole. The work was in part supported by the European Union (MALINV project no 012199). A. Siau was supported by MENRT, Fondation de la Recherche Médicale (FRM), and by the Fondation des Treilles., The authors are grateful to Anne Charlotte Grüner for constructive comments and criticisms of the manuscript and to Emilie Duvaltier, Anna Engström, and Marga van de Vegte-Bolmer for their technical help. Microarray analysis and confocal microscopy were performed using the Pitié-Salpêtrière genomic core facility (P3S) and the Plate-forme d'Imagerie Cellulaire, respectively., Vaquero, Catherine, Institut Pasteur [Paris], Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Muséum national d'Histoire naturelle (MNHN), Service de parasitologie - mycologie [CHU Pitié-Salpétrière], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-CHU Pitié-Salpêtrière [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Sorbonne Université (SU), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU) |
| Jazyk: | angličtina |
| Rok vydání: | 2008 |
| Předmět: |
Plasmodium
Hot Temperature Host cells [SDV]Life Sciences [q-bio] MESH: Malaria Falciparum/metabolism Protozoan Proteins MESH: Plasmodium falciparum/metabolism MESH: Hepatocytes/parasitology Transcriptome MESH: Reverse Transcriptase Polymerase Chain Reaction MESH: Animals Microbiology/Parasitology Malaria Falciparum Biology (General) Cells Cultured Oligonucleotide Array Sequence Analysis Infectivity 0303 health sciences MESH: Plasmodium falciparum/genetics Reverse Transcriptase Polymerase Chain Reaction MESH: Protozoan Proteins/biosynthesis MESH: Gene Expression Profiling/methods MESH: Hepatocytes/metabolism 030302 biochemistry & molecular biology Anopheles MESH: Malaria Falciparum/genetics Genetics and Genomics/Gene Expression Parasitic diseases Up-Regulation 3. Good health Cell biology Pathogenesis and modulation of inflammation [N4i 1] [SDV] Life Sciences [q-bio] medicine.anatomical_structure [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology Sporozoites Hepatocyte MESH: Up-Regulation*/genetics Transcriptome analysis [SDV.MP.PAR] Life Sciences [q-bio]/Microbiology and Parasitology/Parasitology Research Article Infectious Diseases/Tropical and Travel-Associated Diseases MESH: Cells Cultured QH301-705.5 Plasmodium falciparum Immunology MESH: Protozoan Proteins/genetics Biology Salivary glands MESH: Hot Temperature Microbiology MESH: Oligonucleotide Array Sequence Analysis/methods 03 medical and health sciences Virology Genetics medicine Animals Humans [SDV.MP.PAR]Life Sciences [q-bio]/Microbiology and Parasitology/Parasitology Molecular Biology [SDV.MP] Life Sciences [q-bio]/Microbiology and Parasitology 030304 developmental biology MESH: Humans Microarray analysis techniques Gene Expression Profiling Infectious Diseases/Protozoal Infections RC581-607 biology.organism_classification Gene expression profiling Hepatocytes Parasitology Microbial pathogenesis and host defense [UMCN 4.1] Gene expression Immunologic diseases. Allergy Immunity infection and tissue repair [NCMLS 1] |
| Zdroj: | PLoS Pathogens, Vol 4, Iss 8, p e1000121 (2008) PLoS Pathogens PLoS Pathogens, 2008, 4 (8), pp.e1000121. ⟨10.1371/journal.ppat.1000121⟩ PLoS Pathogens, Public Library of Science, 2008, 4 (8), pp.e1000121. ⟨10.1371/journal.ppat.1000121⟩ Plos Pathogens, 4, e1000121-e1000121 Plos Pathogens, 4, 8, pp. e1000121-e1000121 |
| ISSN: | 1553-7374 1553-7366 |
| DOI: | 10.1371/journal.ppat.1000121⟩ |
| Popis: | Plasmodium sporozoites are deposited in the skin by Anopheles mosquitoes. They then find their way to the liver, where they specifically invade hepatocytes in which they develop to yield merozoites infective to red blood cells. Relatively little is known of the molecular interactions during these initial obligatory phases of the infection. Recent data suggested that many of the inoculated sporozoites invade hepatocytes an hour or more after the infective bite. We hypothesised that this pre-invasive period in the mammalian host prepares sporozoites for successful hepatocyte infection. Therefore, the genes whose expression becomes modified prior to hepatocyte invasion would be those likely to code for proteins implicated in the subsequent events of invasion and development. We have used P. falciparum sporozoites and their natural host cells, primary human hepatocytes, in in vitro co-culture system as a model for the pre-invasive period. We first established that under co-culture conditions, sporozoites maintain infectivity for an hour or more, in contrast to a drastic loss in infectivity when hepatocytes were not included. Thus, a differential transcriptome of salivary gland sporozoites versus sporozoites co-cultured with hepatocytes was established using a pan-genomic P. falciparum microarray. The expression of 532 genes was found to have been up-regulated following co-culture. A fifth of these genes had no orthologues in the genomes of Plasmodium species used in rodent models of malaria. Quantitative RT-PCR analysis of a selection of 21 genes confirmed the reliability of the microarray data. Time-course analysis further indicated two patterns of up-regulation following sporozoite co-culture, one transient and the other sustained, suggesting roles in hepatocyte invasion and liver stage development, respectively. This was supported by functional studies of four hitherto uncharacterized proteins of which two were shown to be sporozoite surface proteins involved in hepatocyte invasion, while the other two were predominantly expressed during hepatic parasite development. The genome-wide up-regulation of expression observed supports the hypothesis that the shift from the mosquito to the mammalian host contributes to activate quiescent salivary gland sporozoites into a state of readiness for the hepatic stages. Functional studies on four of the up-regulated genes validated our approach as one means to determine the repertoire of proteins implicated during the early events of the Plasmodium infection, and in this case that of P. falciparum, the species responsible for the severest forms of malaria. Author Summary Sporozoites, the infective form of the malaria parasites Plasmodium, are deposited in the skin by Anopheles mosquitoes. They then find their way to the liver where they specifically invade hepatocytes, in which they develop to yield another form, the merozoite, infective to red blood cells. Relatively little is known of the molecular interactions during these initial obligatory phases of the infection. We studied the changes in gene expression in sporozoites, from the parasite species P. falciparum that infects humans, in an in vitro system where they were co-cultured with their natural host cells, primary human hepatocytes. The whole genome transcriptome profiling carried out led to the identification of 532 genes that were up-regulated following co-culture. This genome-wide up-regulation of expression supports the hypothesis that the shift from the mosquito to the mammalian host contributes to activate quiescent salivary gland sporozoites into a state of readiness for the hepatic stages. Functional studies on four of the up-regulated genes we identified validated our approach as one means to determine the repertoire of proteins implicated during the early events in the infection by P. falciparum, the species responsible for the severest forms of malaria. |
| Databáze: | OpenAIRE |
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