title: Sensing of latent EBV infection through exosomal transfer of 5'pppRNA.
abstract: Complex interactions between DNA herpesviruses and host factors determine the establishment of a life-long asymptomatic latent infection. The lymphotropic Epstein-Barr virus (EBV) seems to avoid recognition by innate sensors despite massive transcription of immunostimulatory small RNAs (EBV-EBERs). Here we demonstrate that in latently infected B cells, EBER1 transcripts interact with the lupus antigen (La) ribonucleoprotein, avoiding cytoplasmic RNA sensors. However, in coculture experiments we observed that latent-infected cells trigger antiviral immunity in dendritic cells (DCs) through selective release and transfer of RNA via exosomes. In ex vivo tonsillar cultures, we observed that EBER1-loaded exosomes are preferentially captured and internalized by human plasmacytoid DCs (pDCs) that express the TIM1 phosphatidylserine receptor, a known viral- and exosomal target. Using an EBER-deficient EBV strain, enzymatic removal of 5'ppp, in vitro transcripts, and coculture experiments, we established that 5'pppEBER1 transfer via exosomes drives antiviral immunity in nonpermissive DCs. Lupus erythematosus patients suffer from elevated EBV load and activated antiviral immunity, in particular in skin lesions that are infiltrated with pDCs. We detected high levels of EBER1 RNA in such skin lesions, as well as EBV-microRNAs, but no intact EBV-DNA, linking non-cell-autonomous EBER1 presence with skin inflammation in predisposed individuals. Collectively, our studies indicate that virus-modified exosomes have a physiological role in the host-pathogen stand-off and may promote inflammatory disease.
authors.affiliation: Department of Pathology, Cancer Center Amsterdam, Vrije Universiteit (VU) University Medical Center, 1081 HV Amsterdam, The Netherlands;
authors.forename: S Rubina
authors.initials: SR
authors.lastname: Baglio
authors.affiliation: Department of Pathology, Cancer Center Amsterdam, Vrije Universiteit (VU) University Medical Center, 1081 HV Amsterdam, The Netherlands;
authors.forename: Monique A J
authors.initials: MA
authors.lastname: van Eijndhoven
authors.affiliation: Department of Pathology, Cancer Center Amsterdam, Vrije Universiteit (VU) University Medical Center, 1081 HV Amsterdam, The Netherlands; Department of Neurosurgery, Cancer Center Amsterdam, VU University Medical Center, 1081 HV Amsterdam, The Netherlands;
authors.forename: Danijela
authors.initials: D
authors.lastname: Koppers-Lalic
authors.affiliation: Department of Neurosurgery, Cancer Center Amsterdam, VU University Medical Center, 1081 HV Amsterdam, The Netherlands;
authors.forename: Jordi
authors.initials: J
authors.lastname: Berenguer
authors.affiliation: Department of Medical Oncology, VU University Medical Center, 1081 HV Amsterdam, The Netherlands;
authors.forename: Sinéad M
authors.initials: SM
authors.lastname: Lougheed
authors.affiliation: Department of Dermatology, Cancer Center Amsterdam, VU University Medical Center, 1081 HV Amsterdam, The Netherlands; Department of Oral Cell Biology, Academic Center for Dentistry Amsterdam, University of Amsterdam and VU University, 1081 HZ Amsterdam, The Netherlands;
authors.forename: Susan
authors.initials: S
authors.lastname: Gibbs
authors.affiliation: Laboratory for Experimental Oncology and Radiobiology, Center for Experimental Molecular Medicine (CEMM), Academic Medical Center (AMC), 1105 AZ Amsterdam, The Netherlands;
authors.forename: Nicolas
authors.initials: N
authors.lastname: Léveillé
authors.affiliation: Department of Otolaryngology, Cancer Center Amsterdam, VU University Medical Center, 1081 HV Amsterdam, The Netherlands;
authors.forename: Rico N P M
authors.initials: RN
authors.lastname: Rinkel
authors.affiliation: Department of Pathology, Cancer Center Amsterdam, Vrije Universiteit (VU) University Medical Center, 1081 HV Amsterdam, The Netherlands;
authors.forename: Erik S
authors.initials: ES
authors.lastname: Hopmans
authors.affiliation: Department of Medicine, University of Utah and George E. Wahlen VA Medical Center, Salt Lake City, UT 84132;
authors.forename: Sankar
authors.initials: S
authors.lastname: Swaminathan
authors.affiliation: Department of Pathology, Cancer Center Amsterdam, Vrije Universiteit (VU) University Medical Center, 1081 HV Amsterdam, The Netherlands;
authors.forename: Sandra A W M
authors.initials: SA
authors.lastname: Verkuijlen
authors.affiliation: Department of Pathology, Cancer Center Amsterdam, Vrije Universiteit (VU) University Medical Center, 1081 HV Amsterdam, The Netherlands;
authors.forename: George L
authors.initials: GL
authors.lastname: Scheffer
authors.affiliation: Department of Infectious Diseases and Immunology, Faculty of Veterinary Medicine, Utrecht University Hospital, 3584 CL Utrecht, The Netherlands;
authors.forename: Frank J M
authors.initials: FJ
authors.lastname: van Kuppeveld
authors.affiliation: Department of Medical Oncology, VU University Medical Center, 1081 HV Amsterdam, The Netherlands;
authors.forename: Tanja D
authors.initials: TD
authors.lastname: de Gruijl
authors.affiliation: Department of Rheumatology, Amsterdam Rheumatology and Immunology Center, VU University Medical Center, 1081 HV Amsterdam, The Netherlands;
authors.forename: Irene E M
authors.initials: IE
authors.lastname: Bultink
authors.affiliation: Department of Obstetrics and Gynecology, Center for Gynaecological Oncology Amsterdam, VU University Medical Center, 1081 HV Amsterdam, The Netherlands;
authors.forename: Ekaterina S
authors.initials: ES
authors.lastname: Jordanova
authors.affiliation: Department of Genetics, University of Granada, 18071 Granada, Spain;
authors.forename: Michael
authors.initials: M
authors.lastname: Hackenberg
authors.affiliation: Department of Medical Oncology, Cancer Center Amsterdam, VU University Medical Center, 1081 HV Amsterdam, The Netherlands.
authors.forename: Sander R
authors.initials: SR
authors.lastname: Piersma
authors.affiliation: Department of Medical Oncology, Cancer Center Amsterdam, VU University Medical Center, 1081 HV Amsterdam, The Netherlands.
authors.forename: Jaco C
authors.initials: JC
authors.lastname: Knol
authors.affiliation: Department of Rheumatology, Amsterdam Rheumatology and Immunology Center, VU University Medical Center, 1081 HV Amsterdam, The Netherlands;
authors.forename: Alexandre E
authors.initials: AE
authors.lastname: Voskuyl
authors.affiliation: Department of Neurosurgery, Cancer Center Amsterdam, VU University Medical Center, 1081 HV Amsterdam, The Netherlands;
authors.forename: Thomas
authors.initials: T
authors.lastname: Wurdinger
authors.affiliation: Department of Medical Oncology, Cancer Center Amsterdam, VU University Medical Center, 1081 HV Amsterdam, The Netherlands.
authors.forename: Connie R
authors.initials: CR
authors.lastname: Jiménez
authors.affiliation: Department of Pathology, Cancer Center Amsterdam, Vrije Universiteit (VU) University Medical Center, 1081 HV Amsterdam, The Netherlands;
authors.forename: Jaap M
authors.initials: JM
authors.lastname: Middeldorp
authors.affiliation: Department of Pathology, Cancer Center Amsterdam, Vrije Universiteit (VU) University Medical Center, 1081 HV Amsterdam, The Netherlands; d.pegtel@vumc.nl.
authors.forename: D Michiel
authors.initials: DM
authors.lastname: Pegtel
mesh_terms: D001692:Biological Transport; D003713:Dendritic Cells / Q000821:virology*; D020031:Epstein-Barr Virus Infections / Q000235:genetics* / Q000276:immunology; D055354:Exosomes / Q000378:metabolism*; D004854:Herpesvirus 4, Human / Q000235:genetics; D006801:Humans; D020543:Proteome; D012367:RNA, Viral / Q000378:metabolism*
chemical_list: C551867:5'PPP-RNA; D020543:Proteome; D012367:RNA, Viral
keywords: EBV-EBER1; dendritic cells; exosomes; innate sensing; skin inflammation