A T164S mutation in the dengue virus NS1 protein is associated with greater disease severity in mice

Kitti Wing Ki Chan, Satoru Watanabe, Jocelyn Y. Jin, Julien Pompon, Don Teng, Sylvie Alonso, Dhanasekaran Vijaykrishna, Scott B. Halstead, Jan K. Marzinek, Peter J. Bond, Bo Burla, Federico Torta, Markus R. Wenk, Eng Eong Ooi, Subhash G. Vasudevan

Research output: Contribution to journalArticleResearchpeer-review

1 Citation (Scopus)

Abstract

Dengue viruses cause severe and sudden human epidemics worldwide. The secreted form of the nonstructural protein 1 (sNS1) of dengue virus causes vascular leakage, a hallmark of severe dengue disease. Here, we reverse engineered the T164S mutation of NS1, associated with the severity of dengue epidemics in the Americas, into a dengue virus serotype 2 mildly infectious strain. The T164S mutant virus decreased infectious virus production and increased sNS1 production in mammalian cell lines and human peripheral blood mononuclear cells (PBMCs) without affecting viral RNA replication. Gene expression profiling of 268 inflammation-associated human genes revealed up-regulation of genes induced in response to vascular leakage. Infection of the mosquito vector Aedes aegypti with the T164S mutant virus resulted in increased viral load in the mosquito midgut and higher sNS1 production compared to wild-type virus infection. Infection of type 1 and 2 interferon receptor-deficient AG129 mice with the T164S mutant virus resulted in severe disease coupled with increased complement activation, tissue inflammation, and more rapid mortality compared to AG129 mice infected with wild-type virus. Molecular dynamics simulations predicted that mutant sNS1 formed stable dimers similar to the wild-type protein, whereas the hexameric mutant sNS1 was predicted to be unstable. Immunoaffinity-purified sNS1 from T164S mutant virus-infected mammalian cells was associated with different lipid classes compared to wild-type sNS1. Treatment of human PBMCs with sNS1 purified from T164S mutant virus resulted in a twofold higher production of proinflammatory cytokines, suggesting a mechanism for how mutant sNS1 may cause more severe dengue disease.

Original languageEnglish
Article numbereaat7726
Number of pages16
JournalScience Translational Medicine
Volume11
Issue number498
DOIs
Publication statusPublished - 26 Jun 2019

Cite this

Chan, Kitti Wing Ki ; Watanabe, Satoru ; Jin, Jocelyn Y. ; Pompon, Julien ; Teng, Don ; Alonso, Sylvie ; Vijaykrishna, Dhanasekaran ; Halstead, Scott B. ; Marzinek, Jan K. ; Bond, Peter J. ; Burla, Bo ; Torta, Federico ; Wenk, Markus R. ; Ooi, Eng Eong ; Vasudevan, Subhash G. / A T164S mutation in the dengue virus NS1 protein is associated with greater disease severity in mice. In: Science Translational Medicine. 2019 ; Vol. 11, No. 498.
@article{a625a774c561450db45bce6509fe78d2,
title = "A T164S mutation in the dengue virus NS1 protein is associated with greater disease severity in mice",
abstract = "Dengue viruses cause severe and sudden human epidemics worldwide. The secreted form of the nonstructural protein 1 (sNS1) of dengue virus causes vascular leakage, a hallmark of severe dengue disease. Here, we reverse engineered the T164S mutation of NS1, associated with the severity of dengue epidemics in the Americas, into a dengue virus serotype 2 mildly infectious strain. The T164S mutant virus decreased infectious virus production and increased sNS1 production in mammalian cell lines and human peripheral blood mononuclear cells (PBMCs) without affecting viral RNA replication. Gene expression profiling of 268 inflammation-associated human genes revealed up-regulation of genes induced in response to vascular leakage. Infection of the mosquito vector Aedes aegypti with the T164S mutant virus resulted in increased viral load in the mosquito midgut and higher sNS1 production compared to wild-type virus infection. Infection of type 1 and 2 interferon receptor-deficient AG129 mice with the T164S mutant virus resulted in severe disease coupled with increased complement activation, tissue inflammation, and more rapid mortality compared to AG129 mice infected with wild-type virus. Molecular dynamics simulations predicted that mutant sNS1 formed stable dimers similar to the wild-type protein, whereas the hexameric mutant sNS1 was predicted to be unstable. Immunoaffinity-purified sNS1 from T164S mutant virus-infected mammalian cells was associated with different lipid classes compared to wild-type sNS1. Treatment of human PBMCs with sNS1 purified from T164S mutant virus resulted in a twofold higher production of proinflammatory cytokines, suggesting a mechanism for how mutant sNS1 may cause more severe dengue disease.",
author = "Chan, {Kitti Wing Ki} and Satoru Watanabe and Jin, {Jocelyn Y.} and Julien Pompon and Don Teng and Sylvie Alonso and Dhanasekaran Vijaykrishna and Halstead, {Scott B.} and Marzinek, {Jan K.} and Bond, {Peter J.} and Bo Burla and Federico Torta and Wenk, {Markus R.} and Ooi, {Eng Eong} and Vasudevan, {Subhash G.}",
year = "2019",
month = "6",
day = "26",
doi = "10.1126/scitranslmed.aat7726",
language = "English",
volume = "11",
journal = "Science Translational Medicine",
issn = "1946-6234",
publisher = "American Association for the Advancement of Science",
number = "498",

}

Chan, KWK, Watanabe, S, Jin, JY, Pompon, J, Teng, D, Alonso, S, Vijaykrishna, D, Halstead, SB, Marzinek, JK, Bond, PJ, Burla, B, Torta, F, Wenk, MR, Ooi, EE & Vasudevan, SG 2019, 'A T164S mutation in the dengue virus NS1 protein is associated with greater disease severity in mice', Science Translational Medicine, vol. 11, no. 498, eaat7726. https://doi.org/10.1126/scitranslmed.aat7726

A T164S mutation in the dengue virus NS1 protein is associated with greater disease severity in mice. / Chan, Kitti Wing Ki; Watanabe, Satoru; Jin, Jocelyn Y.; Pompon, Julien; Teng, Don; Alonso, Sylvie; Vijaykrishna, Dhanasekaran; Halstead, Scott B.; Marzinek, Jan K.; Bond, Peter J.; Burla, Bo; Torta, Federico; Wenk, Markus R.; Ooi, Eng Eong; Vasudevan, Subhash G.

In: Science Translational Medicine, Vol. 11, No. 498, eaat7726, 26.06.2019.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - A T164S mutation in the dengue virus NS1 protein is associated with greater disease severity in mice

AU - Chan, Kitti Wing Ki

AU - Watanabe, Satoru

AU - Jin, Jocelyn Y.

AU - Pompon, Julien

AU - Teng, Don

AU - Alonso, Sylvie

AU - Vijaykrishna, Dhanasekaran

AU - Halstead, Scott B.

AU - Marzinek, Jan K.

AU - Bond, Peter J.

AU - Burla, Bo

AU - Torta, Federico

AU - Wenk, Markus R.

AU - Ooi, Eng Eong

AU - Vasudevan, Subhash G.

PY - 2019/6/26

Y1 - 2019/6/26

N2 - Dengue viruses cause severe and sudden human epidemics worldwide. The secreted form of the nonstructural protein 1 (sNS1) of dengue virus causes vascular leakage, a hallmark of severe dengue disease. Here, we reverse engineered the T164S mutation of NS1, associated with the severity of dengue epidemics in the Americas, into a dengue virus serotype 2 mildly infectious strain. The T164S mutant virus decreased infectious virus production and increased sNS1 production in mammalian cell lines and human peripheral blood mononuclear cells (PBMCs) without affecting viral RNA replication. Gene expression profiling of 268 inflammation-associated human genes revealed up-regulation of genes induced in response to vascular leakage. Infection of the mosquito vector Aedes aegypti with the T164S mutant virus resulted in increased viral load in the mosquito midgut and higher sNS1 production compared to wild-type virus infection. Infection of type 1 and 2 interferon receptor-deficient AG129 mice with the T164S mutant virus resulted in severe disease coupled with increased complement activation, tissue inflammation, and more rapid mortality compared to AG129 mice infected with wild-type virus. Molecular dynamics simulations predicted that mutant sNS1 formed stable dimers similar to the wild-type protein, whereas the hexameric mutant sNS1 was predicted to be unstable. Immunoaffinity-purified sNS1 from T164S mutant virus-infected mammalian cells was associated with different lipid classes compared to wild-type sNS1. Treatment of human PBMCs with sNS1 purified from T164S mutant virus resulted in a twofold higher production of proinflammatory cytokines, suggesting a mechanism for how mutant sNS1 may cause more severe dengue disease.

AB - Dengue viruses cause severe and sudden human epidemics worldwide. The secreted form of the nonstructural protein 1 (sNS1) of dengue virus causes vascular leakage, a hallmark of severe dengue disease. Here, we reverse engineered the T164S mutation of NS1, associated with the severity of dengue epidemics in the Americas, into a dengue virus serotype 2 mildly infectious strain. The T164S mutant virus decreased infectious virus production and increased sNS1 production in mammalian cell lines and human peripheral blood mononuclear cells (PBMCs) without affecting viral RNA replication. Gene expression profiling of 268 inflammation-associated human genes revealed up-regulation of genes induced in response to vascular leakage. Infection of the mosquito vector Aedes aegypti with the T164S mutant virus resulted in increased viral load in the mosquito midgut and higher sNS1 production compared to wild-type virus infection. Infection of type 1 and 2 interferon receptor-deficient AG129 mice with the T164S mutant virus resulted in severe disease coupled with increased complement activation, tissue inflammation, and more rapid mortality compared to AG129 mice infected with wild-type virus. Molecular dynamics simulations predicted that mutant sNS1 formed stable dimers similar to the wild-type protein, whereas the hexameric mutant sNS1 was predicted to be unstable. Immunoaffinity-purified sNS1 from T164S mutant virus-infected mammalian cells was associated with different lipid classes compared to wild-type sNS1. Treatment of human PBMCs with sNS1 purified from T164S mutant virus resulted in a twofold higher production of proinflammatory cytokines, suggesting a mechanism for how mutant sNS1 may cause more severe dengue disease.

UR - http://www.scopus.com/inward/record.url?scp=85068359613&partnerID=8YFLogxK

U2 - 10.1126/scitranslmed.aat7726

DO - 10.1126/scitranslmed.aat7726

M3 - Article

VL - 11

JO - Science Translational Medicine

JF - Science Translational Medicine

SN - 1946-6234

IS - 498

M1 - eaat7726

ER -