Sensing of latent EBV infection through exosomal transfer of 5'pppRNA

S. Rubina Baglio, Monique A. J. van Eijndhoven, Danijela Koppers-Lalic, Jordi Berenguer, Sinead M. Lougheed, Susan Gibbs, Nicolas Leveille, Rico N. P. M. Rinkel, Erik S. Hopmans, Sankar Swaminathan, Sandra A. W. M. Verkuijlen, George L. Scheffer, Frank J. M. van Kuppeveld, Tanja D. de Gruijl, Irene E. M. Bultink, Ekaterina S. Jordanova, Michael Hackenberg, Sander R. Piersma, Jaco C. Knol, Alexandre E. Voskuyl & 4 others Thomas Wurdinger, Connie R. Jimenez, Jaap M. Middeldorp, D. Michiel Pegtel

Research output: Contribution to journalArticleAcademicpeer-review

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.

Original languageEnglish
Pages (from-to)E587-E596
JournalProceedings of the National Academy of Sciences of the United States of America
Volume113
Issue number5
DOIs
Publication statusPublished - 2 Feb 2016

Cite this

@article{ec8f8235c0434f37abfec581460072f7,
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.",
keywords = "Biological Transport, Dendritic Cells, Epstein-Barr Virus Infections, Exosomes, Herpesvirus 4, Human, Humans, Proteome, RNA, Viral, Journal Article, Research Support, Non-U.S. Gov't",
author = "Baglio, {S. Rubina} and {van Eijndhoven}, {Monique A. J.} and Danijela Koppers-Lalic and Jordi Berenguer and Lougheed, {Sinead M.} and Susan Gibbs and Nicolas Leveille and Rinkel, {Rico N. P. M.} and Hopmans, {Erik S.} and Sankar Swaminathan and Verkuijlen, {Sandra A. W. M.} and Scheffer, {George L.} and {van Kuppeveld}, {Frank J. M.} and {de Gruijl}, {Tanja D.} and Bultink, {Irene E. M.} and Jordanova, {Ekaterina S.} and Michael Hackenberg and Piersma, {Sander R.} and Knol, {Jaco C.} and Voskuyl, {Alexandre E.} and Thomas Wurdinger and Jimenez, {Connie R.} and Middeldorp, {Jaap M.} and Pegtel, {D. Michiel}",
year = "2016",
month = "2",
day = "2",
doi = "10.1073/pnas.1518130113",
language = "English",
volume = "113",
pages = "E587--E596",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
number = "5",

}

Sensing of latent EBV infection through exosomal transfer of 5'pppRNA. / Baglio, S. Rubina; van Eijndhoven, Monique A. J.; Koppers-Lalic, Danijela; Berenguer, Jordi; Lougheed, Sinead M.; Gibbs, Susan; Leveille, Nicolas; Rinkel, Rico N. P. M.; Hopmans, Erik S.; Swaminathan, Sankar; Verkuijlen, Sandra A. W. M.; Scheffer, George L.; van Kuppeveld, Frank J. M.; de Gruijl, Tanja D.; Bultink, Irene E. M.; Jordanova, Ekaterina S.; Hackenberg, Michael; Piersma, Sander R.; Knol, Jaco C.; Voskuyl, Alexandre E.; Wurdinger, Thomas; Jimenez, Connie R.; Middeldorp, Jaap M.; Pegtel, D. Michiel.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 113, No. 5, 02.02.2016, p. E587-E596.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Sensing of latent EBV infection through exosomal transfer of 5'pppRNA

AU - Baglio, S. Rubina

AU - van Eijndhoven, Monique A. J.

AU - Koppers-Lalic, Danijela

AU - Berenguer, Jordi

AU - Lougheed, Sinead M.

AU - Gibbs, Susan

AU - Leveille, Nicolas

AU - Rinkel, Rico N. P. M.

AU - Hopmans, Erik S.

AU - Swaminathan, Sankar

AU - Verkuijlen, Sandra A. W. M.

AU - Scheffer, George L.

AU - van Kuppeveld, Frank J. M.

AU - de Gruijl, Tanja D.

AU - Bultink, Irene E. M.

AU - Jordanova, Ekaterina S.

AU - Hackenberg, Michael

AU - Piersma, Sander R.

AU - Knol, Jaco C.

AU - Voskuyl, Alexandre E.

AU - Wurdinger, Thomas

AU - Jimenez, Connie R.

AU - Middeldorp, Jaap M.

AU - Pegtel, D. Michiel

PY - 2016/2/2

Y1 - 2016/2/2

N2 - 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.

AB - 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.

KW - Biological Transport

KW - Dendritic Cells

KW - Epstein-Barr Virus Infections

KW - Exosomes

KW - Herpesvirus 4, Human

KW - Humans

KW - Proteome

KW - RNA, Viral

KW - Journal Article

KW - Research Support, Non-U.S. Gov't

U2 - 10.1073/pnas.1518130113

DO - 10.1073/pnas.1518130113

M3 - Article

VL - 113

SP - E587-E596

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

SN - 0027-8424

IS - 5

ER -