Epstein-Barr virus (EBV) preferentially infects epithelial cells and B lymphocytes and sometimes T and NK lymphocytes. Persistence of EBV-infected cells results in severe lymphoproliferative disorders (LPDs). Diagnosis of EBV-driven T or NK cell LPD and chronic active EBV diseases (CAEBV) is difficult, often requiring biopsies. Herein, we report a flow-FISH cytometry assay that detects cells expressing EBV-encoded small RNAs (EBERs), allowing rapid identification of EBV-infected cells among PBMCs. EBV-infected B, T, and/or NK cells were detectable in various LPD conditions. Diagnosis of CAEBV in 22 patients of Caucasian and African origins was established. All exhibited circulating EBV-infected T and/or NK cells, highlighting that CAEBV is not restricted to native American and Asian populations. Proportions of EBV-infected cells correlated with blood EBV loads. We showed that EBV-infected T cells had an effector memory activated phenotype, whereas EBV-infected B cells expressed plasma cell differentiation markers. Thus, this method achieves accurate and unambiguous diagnoses of different forms of EBV-driven LPD and represents a powerful tool to study their pathophysiological mechanisms.
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2 November 2020
Technical Advances and Resources|
August 18 2020
Rapid identification and characterization of infected cells in blood during chronic active Epstein-Barr virus infection
In Special Collection:
2020 Clinical Collection on Viral Infections
Benjamin Fournier,
Benjamin Fournier
Conceptualization, Formal analysis, Investigation, Methodology, Visualization, Writing - original draft
1
Laboratory of Lymphocyte Activation and Susceptibility to EBV Infection, Institut National de la Santé et de la Recherche Médicale UMR 1163, Imagine Institute, Paris, France
2
Université de Paris, Paris, France
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David Boutboul,
David Boutboul
Investigation, Resources, Writing - review & editing
3
Department of Clinical Immunology, Saint-Louis Hospital, Assistance Publique–Hôpitaux de Paris, Paris, France
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Julie Bruneau,
Julie Bruneau
Conceptualization, Data curation, Investigation, Resources, Validation, Visualization, Writing - original draft, Writing - review & editing
2
Université de Paris, Paris, France
4
Department of Pathology, Necker-Enfants Malades Hospital, Assistance Publique–Hôpitaux de Paris, Paris, France
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Charline Miot,
Charline Miot
Investigation
5
Department of Pediatric Immunology Hematology and Oncology, University Hospital, Angers, France
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Cécile Boulanger,
Cécile Boulanger
Writing - review & editing
6
Institut Roi Albert II, Cancerology and Hematology Departments, University Clinics Saint-Luc Hospital, Brussels, Belgium
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Marion Malphettes,
Marion Malphettes
Resources
3
Department of Clinical Immunology, Saint-Louis Hospital, Assistance Publique–Hôpitaux de Paris, Paris, France
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Isabelle Pellier,
Isabelle Pellier
Conceptualization, Resources, Writing - review & editing
5
Department of Pediatric Immunology Hematology and Oncology, University Hospital, Angers, France
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Bertrand Dunogué,
Bertrand Dunogué
Investigation, Resources, Writing - review & editing
7
Department of Internal Medicine, Cochin Hospital, National Referral Centre for Systemic and Autoimmune Diseases, Cochin Hospital, Assistance Publique–Hôpitaux de Paris, Paris, France
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Benjamin Terrier,
Benjamin Terrier
Resources, Writing - review & editing
7
Department of Internal Medicine, Cochin Hospital, National Referral Centre for Systemic and Autoimmune Diseases, Cochin Hospital, Assistance Publique–Hôpitaux de Paris, Paris, France
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Felipe Suarez,
Felipe Suarez
Resources
8
Department of Adult Hematology, Necker-Enfants Malades Hospital, Assistance Publique–Hôpitaux de Paris, Paris, France
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Stéphane Blanche,
Stéphane Blanche
Resources
9
Department of Pediatric Immunology, Hematology and Rheumatology, Necker-Enfants Malades Hospital, Assistance Publique–Hôpitaux de Paris, Paris, France
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Martin Castelle,
Martin Castelle
Resources, Writing - review & editing
9
Department of Pediatric Immunology, Hematology and Rheumatology, Necker-Enfants Malades Hospital, Assistance Publique–Hôpitaux de Paris, Paris, France
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Sarah Winter,
Sarah Winter
Investigation, Resources, Writing - review & editing
9
Department of Pediatric Immunology, Hematology and Rheumatology, Necker-Enfants Malades Hospital, Assistance Publique–Hôpitaux de Paris, Paris, France
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Henri-Jacques Delecluse,
Henri-Jacques Delecluse
Resources
10
Unit F100, Institut National de la Santé et de la Recherche Médicale U1074, Deutsches Krebsforschungszentrum, German Cancer Research Center, Heidelberg, Germany
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Thierry Molina,
Thierry Molina
Data curation, Resources, Validation
2
Université de Paris, Paris, France
4
Department of Pathology, Necker-Enfants Malades Hospital, Assistance Publique–Hôpitaux de Paris, Paris, France
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Capucine Picard,
Capucine Picard
Supervision, Validation, Writing - original draft
1
Laboratory of Lymphocyte Activation and Susceptibility to EBV Infection, Institut National de la Santé et de la Recherche Médicale UMR 1163, Imagine Institute, Paris, France
2
Université de Paris, Paris, France
11
Study Center for Primary Immunodeficiencies, Necker-Enfants Malades Hospital, Assistance Publique–Hôpitaux de Paris, Paris, France
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Stephan Ehl,
Stephan Ehl
Conceptualization, Funding acquisition, Project administration, Supervision, Validation, Visualization, Writing - review & editing
12
Institute for Immunodeficiency-Center for Chronic Immunodeficiency, Department of Pediatrics and Adolescent Medicine, Medical Center - Faculty of Medicine, University of Freiburg, Germany
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Despina Moshous,
Despina Moshous
Resources, Supervision, Writing - review & editing
2
Université de Paris, Paris, France
9
Department of Pediatric Immunology, Hematology and Rheumatology, Necker-Enfants Malades Hospital, Assistance Publique–Hôpitaux de Paris, Paris, France
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Lionel Galicier,
Lionel Galicier
Resources, Writing - review & editing
3
Department of Clinical Immunology, Saint-Louis Hospital, Assistance Publique–Hôpitaux de Paris, Paris, France
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Vincent Barlogis,
Vincent Barlogis
Resources, Writing - review & editing
13
Department of Pediatric Hematology-Oncology, La Timone Hospital, Marseille, France
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Alain Fischer,
Alain Fischer
Supervision, Writing - review & editing
2
Université de Paris, Paris, France
9
Department of Pediatric Immunology, Hematology and Rheumatology, Necker-Enfants Malades Hospital, Assistance Publique–Hôpitaux de Paris, Paris, France
14
Collège de France, Paris, France
15
Institut National de la Santé et de la Recherche Médicale UMR 1163, Paris, France
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Bénédicte Neven,
Bénédicte Neven
Conceptualization, Resources, Writing - review & editing
2
Université de Paris, Paris, France
9
Department of Pediatric Immunology, Hematology and Rheumatology, Necker-Enfants Malades Hospital, Assistance Publique–Hôpitaux de Paris, Paris, France
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Sylvain Latour
Conceptualization, Funding acquisition, Supervision, Validation, Visualization, Writing - original draft
1
Laboratory of Lymphocyte Activation and Susceptibility to EBV Infection, Institut National de la Santé et de la Recherche Médicale UMR 1163, Imagine Institute, Paris, France
2
Université de Paris, Paris, France
Correspondence to Sylvain Latour: sylvain.latour@inserm.fr
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Benjamin Fournier
Conceptualization, Formal analysis, Investigation, Methodology, Visualization, Writing - original draft
1
Laboratory of Lymphocyte Activation and Susceptibility to EBV Infection, Institut National de la Santé et de la Recherche Médicale UMR 1163, Imagine Institute, Paris, France
2
Université de Paris, Paris, France
David Boutboul
Investigation, Resources, Writing - review & editing
3
Department of Clinical Immunology, Saint-Louis Hospital, Assistance Publique–Hôpitaux de Paris, Paris, France
Julie Bruneau
Conceptualization, Data curation, Investigation, Resources, Validation, Visualization, Writing - original draft, Writing - review & editing
2
Université de Paris, Paris, France
4
Department of Pathology, Necker-Enfants Malades Hospital, Assistance Publique–Hôpitaux de Paris, Paris, France
Charline Miot
Investigation
5
Department of Pediatric Immunology Hematology and Oncology, University Hospital, Angers, France
Cécile Boulanger
Writing - review & editing
6
Institut Roi Albert II, Cancerology and Hematology Departments, University Clinics Saint-Luc Hospital, Brussels, Belgium
Marion Malphettes
Resources
3
Department of Clinical Immunology, Saint-Louis Hospital, Assistance Publique–Hôpitaux de Paris, Paris, France
Isabelle Pellier
Conceptualization, Resources, Writing - review & editing
5
Department of Pediatric Immunology Hematology and Oncology, University Hospital, Angers, France
Bertrand Dunogué
Investigation, Resources, Writing - review & editing
7
Department of Internal Medicine, Cochin Hospital, National Referral Centre for Systemic and Autoimmune Diseases, Cochin Hospital, Assistance Publique–Hôpitaux de Paris, Paris, France
Benjamin Terrier
Resources, Writing - review & editing
7
Department of Internal Medicine, Cochin Hospital, National Referral Centre for Systemic and Autoimmune Diseases, Cochin Hospital, Assistance Publique–Hôpitaux de Paris, Paris, France
Felipe Suarez
Resources
8
Department of Adult Hematology, Necker-Enfants Malades Hospital, Assistance Publique–Hôpitaux de Paris, Paris, France
Stéphane Blanche
Resources
9
Department of Pediatric Immunology, Hematology and Rheumatology, Necker-Enfants Malades Hospital, Assistance Publique–Hôpitaux de Paris, Paris, France
Martin Castelle
Resources, Writing - review & editing
9
Department of Pediatric Immunology, Hematology and Rheumatology, Necker-Enfants Malades Hospital, Assistance Publique–Hôpitaux de Paris, Paris, France
Sarah Winter
Investigation, Resources, Writing - review & editing
9
Department of Pediatric Immunology, Hematology and Rheumatology, Necker-Enfants Malades Hospital, Assistance Publique–Hôpitaux de Paris, Paris, France
Henri-Jacques Delecluse
Resources
10
Unit F100, Institut National de la Santé et de la Recherche Médicale U1074, Deutsches Krebsforschungszentrum, German Cancer Research Center, Heidelberg, Germany
Thierry Molina
Data curation, Resources, Validation
2
Université de Paris, Paris, France
4
Department of Pathology, Necker-Enfants Malades Hospital, Assistance Publique–Hôpitaux de Paris, Paris, France
Capucine Picard
Supervision, Validation, Writing - original draft
1
Laboratory of Lymphocyte Activation and Susceptibility to EBV Infection, Institut National de la Santé et de la Recherche Médicale UMR 1163, Imagine Institute, Paris, France
2
Université de Paris, Paris, France
11
Study Center for Primary Immunodeficiencies, Necker-Enfants Malades Hospital, Assistance Publique–Hôpitaux de Paris, Paris, France
Stephan Ehl
Conceptualization, Funding acquisition, Project administration, Supervision, Validation, Visualization, Writing - review & editing
12
Institute for Immunodeficiency-Center for Chronic Immunodeficiency, Department of Pediatrics and Adolescent Medicine, Medical Center - Faculty of Medicine, University of Freiburg, Germany
Despina Moshous
Resources, Supervision, Writing - review & editing
2
Université de Paris, Paris, France
9
Department of Pediatric Immunology, Hematology and Rheumatology, Necker-Enfants Malades Hospital, Assistance Publique–Hôpitaux de Paris, Paris, France
Lionel Galicier
Resources, Writing - review & editing
3
Department of Clinical Immunology, Saint-Louis Hospital, Assistance Publique–Hôpitaux de Paris, Paris, France
Vincent Barlogis
Resources, Writing - review & editing
13
Department of Pediatric Hematology-Oncology, La Timone Hospital, Marseille, France
Alain Fischer
Supervision, Writing - review & editing
2
Université de Paris, Paris, France
9
Department of Pediatric Immunology, Hematology and Rheumatology, Necker-Enfants Malades Hospital, Assistance Publique–Hôpitaux de Paris, Paris, France
14
Collège de France, Paris, France
15
Institut National de la Santé et de la Recherche Médicale UMR 1163, Paris, France
Bénédicte Neven
Conceptualization, Resources, Writing - review & editing
2
Université de Paris, Paris, France
9
Department of Pediatric Immunology, Hematology and Rheumatology, Necker-Enfants Malades Hospital, Assistance Publique–Hôpitaux de Paris, Paris, France
Sylvain Latour
Conceptualization, Funding acquisition, Supervision, Validation, Visualization, Writing - original draft
1
Laboratory of Lymphocyte Activation and Susceptibility to EBV Infection, Institut National de la Santé et de la Recherche Médicale UMR 1163, Imagine Institute, Paris, France
2
Université de Paris, Paris, France
Correspondence to Sylvain Latour: sylvain.latour@inserm.fr
Received:
November 30 2019
Revision Received:
May 29 2020
Accepted:
July 09 2020
Online Issn: 1540-9538
Print Issn: 0022-1007
Funding:
Fondation pour la Recherche Médicale
(FDM20170638301)
Institut National de la Santé et de la Recherche Médicale
(NO AWARD)
Ligue Contre le Cancer
(NO AWARD)
© 2020 Fournier et al.
2020
This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms/). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 International license, as described at https://creativecommons.org/licenses/by-nc-sa/4.0/).
J Exp Med (2020) 217 (11): e20192262.
Article history
Received:
November 30 2019
Revision Received:
May 29 2020
Accepted:
July 09 2020
Citation
Benjamin Fournier, David Boutboul, Julie Bruneau, Charline Miot, Cécile Boulanger, Marion Malphettes, Isabelle Pellier, Bertrand Dunogué, Benjamin Terrier, Felipe Suarez, Stéphane Blanche, Martin Castelle, Sarah Winter, Henri-Jacques Delecluse, Thierry Molina, Capucine Picard, Stephan Ehl, Despina Moshous, Lionel Galicier, Vincent Barlogis, Alain Fischer, Bénédicte Neven, Sylvain Latour; Rapid identification and characterization of infected cells in blood during chronic active Epstein-Barr virus infection. J Exp Med 2 November 2020; 217 (11): e20192262. doi: https://doi.org/10.1084/jem.20192262
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