Ebola virus binding to Tim-1 on T lymphocytes induces a cytokine storm

Patrick Younan, Mathieu Iampietro, Andrew Nishida, Palaniappan Ramanathan, Rodrigo I. Santos, Mukta Dutta, Ndongala Michel Lubaki, Richard A. Koup, Michael G. Katze, Alexander Bukreyev

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Ebola virus (EBOV) disease (EVD) results from an exacerbated immunological response that is highlighted by a burst in the production of inflammatory mediators known as a “cytokine storm.” Previous reports have suggested that non-specific activation of T lymphocytes may play a central role in this phenomenon. T-cell immunoglobulin and mucin domain-containing protein 1 (Tim-1) has recently been shown to interact with virion-associated phosphatidylserine to promote infection. Here, we demonstrate the central role of Tim-1 in EBOV pathogenesis, as Tim-1-/- mice exhibited increased survival rates and reduced disease severity; surprisingly, only a limited decrease in viremia was detected. Tim-1-/- mice exhibited a modified inflammatory response as evidenced by changes in serum cytokines and activation of T helper subsets. A series of in vitro assays based on the Tim-1 expression profile on T cells demonstrated that despite the apparent absence of detectable viral replication in T lymphocytes, EBOV directly binds to isolated T lymphocytes in a phosphatidylserine–Tim-1-dependent manner. Exposure to EBOV resulted in the rapid development of a CD4Hi CD3Low population, non-antigen-specific activation, and cytokine production. Transcriptome and Western blot analysis of EBOV-stimulated CD4+ T cells confirmed the induction of the Tim-1 signaling pathway. Furthermore, comparative analysis of transcriptome data and cytokine/chemokine analysis of supernatants highlight the similarities associated with EBOV-stimulated T cells and the onset of a cytokine storm. Flow cytometry revealed virtually exclusive binding and activation of central memory CD4+ T cells. These findings provide evidence for the role of Tim-1 in the induction of a cytokine storm phenomenon and the pathogenesis of EVD. IMPORTANCE Ebola virus infection is characterized by a massive release of inflammatory mediators, which has come to be known as a cytokine storm. The severity of the cytokine storm is consistently linked with fatal disease outcome. Previous findings have demonstrated that specific T-cell subsets are key contributors to the onset of a cytokine storm. In this study, we investigated the role of Tim-1, a T-cell-receptor-independent trigger of T-cell activation. We first demonstrated that Tim-1-knockout (KO) mice survive lethal Ebola virus challenge. We then used a series of in vitro assays to demonstrate that Ebola virus directly binds primary T cells in a Tim-1– phosphatidylserine-dependent manner. We noted that binding induces a cytokine storm-like phenomenon and that blocking Tim-1–phosphatidylserine interactions reduces viral binding, T-cell activation, and cytokine production. These findings high- light a previously unknown role of Tim-1 in the development of a cytokine storm and “immune paralysis.”.

Original languageEnglish (US)
Article numbere00845-17
JournalmBio
Volume8
Issue number5
DOIs
StatePublished - Sep 1 2017

Fingerprint

Ebolavirus
Virus Attachment
Cytokines
T-Lymphocytes
Ebola Hemorrhagic Fever
Phosphatidylserines
Fatal Outcome
Viremia
T-Lymphocyte Subsets
Gene Expression Profiling
T-Cell Antigen Receptor
Transcriptome
Chemokines
Knockout Mice
Paralysis
Virion

Keywords

  • Cytokine storm
  • Cytokines
  • Ebola virus
  • T lymphocytes
  • Transcriptome
  • Viral pathogenesis

ASJC Scopus subject areas

  • Microbiology
  • Virology

Cite this

Younan, P., Iampietro, M., Nishida, A., Ramanathan, P., Santos, R. I., Dutta, M., ... Bukreyev, A. (2017). Ebola virus binding to Tim-1 on T lymphocytes induces a cytokine storm. mBio, 8(5), [e00845-17]. https://doi.org/10.1128/mBio.00845-17

Ebola virus binding to Tim-1 on T lymphocytes induces a cytokine storm. / Younan, Patrick; Iampietro, Mathieu; Nishida, Andrew; Ramanathan, Palaniappan; Santos, Rodrigo I.; Dutta, Mukta; Lubaki, Ndongala Michel; Koup, Richard A.; Katze, Michael G.; Bukreyev, Alexander.

In: mBio, Vol. 8, No. 5, e00845-17, 01.09.2017.

Research output: Contribution to journalArticle

Younan, P, Iampietro, M, Nishida, A, Ramanathan, P, Santos, RI, Dutta, M, Lubaki, NM, Koup, RA, Katze, MG & Bukreyev, A 2017, 'Ebola virus binding to Tim-1 on T lymphocytes induces a cytokine storm', mBio, vol. 8, no. 5, e00845-17. https://doi.org/10.1128/mBio.00845-17
Younan P, Iampietro M, Nishida A, Ramanathan P, Santos RI, Dutta M et al. Ebola virus binding to Tim-1 on T lymphocytes induces a cytokine storm. mBio. 2017 Sep 1;8(5). e00845-17. https://doi.org/10.1128/mBio.00845-17
Younan, Patrick ; Iampietro, Mathieu ; Nishida, Andrew ; Ramanathan, Palaniappan ; Santos, Rodrigo I. ; Dutta, Mukta ; Lubaki, Ndongala Michel ; Koup, Richard A. ; Katze, Michael G. ; Bukreyev, Alexander. / Ebola virus binding to Tim-1 on T lymphocytes induces a cytokine storm. In: mBio. 2017 ; Vol. 8, No. 5.
@article{fd4f3b26c7e94807a816ba2f17c114f8,
title = "Ebola virus binding to Tim-1 on T lymphocytes induces a cytokine storm",
abstract = "Ebola virus (EBOV) disease (EVD) results from an exacerbated immunological response that is highlighted by a burst in the production of inflammatory mediators known as a “cytokine storm.” Previous reports have suggested that non-specific activation of T lymphocytes may play a central role in this phenomenon. T-cell immunoglobulin and mucin domain-containing protein 1 (Tim-1) has recently been shown to interact with virion-associated phosphatidylserine to promote infection. Here, we demonstrate the central role of Tim-1 in EBOV pathogenesis, as Tim-1-/- mice exhibited increased survival rates and reduced disease severity; surprisingly, only a limited decrease in viremia was detected. Tim-1-/- mice exhibited a modified inflammatory response as evidenced by changes in serum cytokines and activation of T helper subsets. A series of in vitro assays based on the Tim-1 expression profile on T cells demonstrated that despite the apparent absence of detectable viral replication in T lymphocytes, EBOV directly binds to isolated T lymphocytes in a phosphatidylserine–Tim-1-dependent manner. Exposure to EBOV resulted in the rapid development of a CD4Hi CD3Low population, non-antigen-specific activation, and cytokine production. Transcriptome and Western blot analysis of EBOV-stimulated CD4+ T cells confirmed the induction of the Tim-1 signaling pathway. Furthermore, comparative analysis of transcriptome data and cytokine/chemokine analysis of supernatants highlight the similarities associated with EBOV-stimulated T cells and the onset of a cytokine storm. Flow cytometry revealed virtually exclusive binding and activation of central memory CD4+ T cells. These findings provide evidence for the role of Tim-1 in the induction of a cytokine storm phenomenon and the pathogenesis of EVD. IMPORTANCE Ebola virus infection is characterized by a massive release of inflammatory mediators, which has come to be known as a cytokine storm. The severity of the cytokine storm is consistently linked with fatal disease outcome. Previous findings have demonstrated that specific T-cell subsets are key contributors to the onset of a cytokine storm. In this study, we investigated the role of Tim-1, a T-cell-receptor-independent trigger of T-cell activation. We first demonstrated that Tim-1-knockout (KO) mice survive lethal Ebola virus challenge. We then used a series of in vitro assays to demonstrate that Ebola virus directly binds primary T cells in a Tim-1– phosphatidylserine-dependent manner. We noted that binding induces a cytokine storm-like phenomenon and that blocking Tim-1–phosphatidylserine interactions reduces viral binding, T-cell activation, and cytokine production. These findings high- light a previously unknown role of Tim-1 in the development of a cytokine storm and “immune paralysis.”.",
keywords = "Cytokine storm, Cytokines, Ebola virus, T lymphocytes, Transcriptome, Viral pathogenesis",
author = "Patrick Younan and Mathieu Iampietro and Andrew Nishida and Palaniappan Ramanathan and Santos, {Rodrigo I.} and Mukta Dutta and Lubaki, {Ndongala Michel} and Koup, {Richard A.} and Katze, {Michael G.} and Alexander Bukreyev",
year = "2017",
month = "9",
day = "1",
doi = "10.1128/mBio.00845-17",
language = "English (US)",
volume = "8",
journal = "mBio",
issn = "2161-2129",
publisher = "American Society for Microbiology",
number = "5",

}

TY - JOUR

T1 - Ebola virus binding to Tim-1 on T lymphocytes induces a cytokine storm

AU - Younan, Patrick

AU - Iampietro, Mathieu

AU - Nishida, Andrew

AU - Ramanathan, Palaniappan

AU - Santos, Rodrigo I.

AU - Dutta, Mukta

AU - Lubaki, Ndongala Michel

AU - Koup, Richard A.

AU - Katze, Michael G.

AU - Bukreyev, Alexander

PY - 2017/9/1

Y1 - 2017/9/1

N2 - Ebola virus (EBOV) disease (EVD) results from an exacerbated immunological response that is highlighted by a burst in the production of inflammatory mediators known as a “cytokine storm.” Previous reports have suggested that non-specific activation of T lymphocytes may play a central role in this phenomenon. T-cell immunoglobulin and mucin domain-containing protein 1 (Tim-1) has recently been shown to interact with virion-associated phosphatidylserine to promote infection. Here, we demonstrate the central role of Tim-1 in EBOV pathogenesis, as Tim-1-/- mice exhibited increased survival rates and reduced disease severity; surprisingly, only a limited decrease in viremia was detected. Tim-1-/- mice exhibited a modified inflammatory response as evidenced by changes in serum cytokines and activation of T helper subsets. A series of in vitro assays based on the Tim-1 expression profile on T cells demonstrated that despite the apparent absence of detectable viral replication in T lymphocytes, EBOV directly binds to isolated T lymphocytes in a phosphatidylserine–Tim-1-dependent manner. Exposure to EBOV resulted in the rapid development of a CD4Hi CD3Low population, non-antigen-specific activation, and cytokine production. Transcriptome and Western blot analysis of EBOV-stimulated CD4+ T cells confirmed the induction of the Tim-1 signaling pathway. Furthermore, comparative analysis of transcriptome data and cytokine/chemokine analysis of supernatants highlight the similarities associated with EBOV-stimulated T cells and the onset of a cytokine storm. Flow cytometry revealed virtually exclusive binding and activation of central memory CD4+ T cells. These findings provide evidence for the role of Tim-1 in the induction of a cytokine storm phenomenon and the pathogenesis of EVD. IMPORTANCE Ebola virus infection is characterized by a massive release of inflammatory mediators, which has come to be known as a cytokine storm. The severity of the cytokine storm is consistently linked with fatal disease outcome. Previous findings have demonstrated that specific T-cell subsets are key contributors to the onset of a cytokine storm. In this study, we investigated the role of Tim-1, a T-cell-receptor-independent trigger of T-cell activation. We first demonstrated that Tim-1-knockout (KO) mice survive lethal Ebola virus challenge. We then used a series of in vitro assays to demonstrate that Ebola virus directly binds primary T cells in a Tim-1– phosphatidylserine-dependent manner. We noted that binding induces a cytokine storm-like phenomenon and that blocking Tim-1–phosphatidylserine interactions reduces viral binding, T-cell activation, and cytokine production. These findings high- light a previously unknown role of Tim-1 in the development of a cytokine storm and “immune paralysis.”.

AB - Ebola virus (EBOV) disease (EVD) results from an exacerbated immunological response that is highlighted by a burst in the production of inflammatory mediators known as a “cytokine storm.” Previous reports have suggested that non-specific activation of T lymphocytes may play a central role in this phenomenon. T-cell immunoglobulin and mucin domain-containing protein 1 (Tim-1) has recently been shown to interact with virion-associated phosphatidylserine to promote infection. Here, we demonstrate the central role of Tim-1 in EBOV pathogenesis, as Tim-1-/- mice exhibited increased survival rates and reduced disease severity; surprisingly, only a limited decrease in viremia was detected. Tim-1-/- mice exhibited a modified inflammatory response as evidenced by changes in serum cytokines and activation of T helper subsets. A series of in vitro assays based on the Tim-1 expression profile on T cells demonstrated that despite the apparent absence of detectable viral replication in T lymphocytes, EBOV directly binds to isolated T lymphocytes in a phosphatidylserine–Tim-1-dependent manner. Exposure to EBOV resulted in the rapid development of a CD4Hi CD3Low population, non-antigen-specific activation, and cytokine production. Transcriptome and Western blot analysis of EBOV-stimulated CD4+ T cells confirmed the induction of the Tim-1 signaling pathway. Furthermore, comparative analysis of transcriptome data and cytokine/chemokine analysis of supernatants highlight the similarities associated with EBOV-stimulated T cells and the onset of a cytokine storm. Flow cytometry revealed virtually exclusive binding and activation of central memory CD4+ T cells. These findings provide evidence for the role of Tim-1 in the induction of a cytokine storm phenomenon and the pathogenesis of EVD. IMPORTANCE Ebola virus infection is characterized by a massive release of inflammatory mediators, which has come to be known as a cytokine storm. The severity of the cytokine storm is consistently linked with fatal disease outcome. Previous findings have demonstrated that specific T-cell subsets are key contributors to the onset of a cytokine storm. In this study, we investigated the role of Tim-1, a T-cell-receptor-independent trigger of T-cell activation. We first demonstrated that Tim-1-knockout (KO) mice survive lethal Ebola virus challenge. We then used a series of in vitro assays to demonstrate that Ebola virus directly binds primary T cells in a Tim-1– phosphatidylserine-dependent manner. We noted that binding induces a cytokine storm-like phenomenon and that blocking Tim-1–phosphatidylserine interactions reduces viral binding, T-cell activation, and cytokine production. These findings high- light a previously unknown role of Tim-1 in the development of a cytokine storm and “immune paralysis.”.

KW - Cytokine storm

KW - Cytokines

KW - Ebola virus

KW - T lymphocytes

KW - Transcriptome

KW - Viral pathogenesis

UR - http://www.scopus.com/inward/record.url?scp=85033730437&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85033730437&partnerID=8YFLogxK

U2 - 10.1128/mBio.00845-17

DO - 10.1128/mBio.00845-17

M3 - Article

VL - 8

JO - mBio

JF - mBio

SN - 2161-2129

IS - 5

M1 - e00845-17

ER -