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  1. Article ; Online: Dengue virus induces PCSK9 expression to alter antiviral responses and disease outcomes.

    Gan, Esther Shuyi / Tan, Hwee Cheng / Le, Duyen Huynh Thi / Huynh, Trieu Trung / Wills, Bridget / Seidah, Nabil G / Ooi, Eng Eong / Yacoub, Sophie

    The Journal of clinical investigation

    2020  Volume 130, Issue 10, Page(s) 5223–5234

    Abstract: Dengue virus (DENV) infection requires cholesterol as a proviral factor, although statin treatment did not show antiviral efficacy in patients with dengue. Here, we show that DENV infection manipulated cholesterol metabolism in cells residing in low- ... ...

    Abstract Dengue virus (DENV) infection requires cholesterol as a proviral factor, although statin treatment did not show antiviral efficacy in patients with dengue. Here, we show that DENV infection manipulated cholesterol metabolism in cells residing in low-oxygen microenvironments (hypoxia) such as in the liver, spleen, and lymph nodes. DENV infection induced expression of proprotein convertase subtilisin/kexin type 9 (PCSK9), which reduces low-density lipoprotein receptor (LDLR) recycling and hence cholesterol uptake. We found that, whereas LDLR uptake would have distributed cholesterol throughout the various cell compartments, de novo cholesterol synthesis enriched this lipid in the endoplasmic reticulum (ER). With cholesterol enrichment in the ER, ER-resident STING and type I IFN (IFN) activation was repressed during DENV infection. Our in vitro findings were further supported by the detection of elevated plasma PCSK9 levels in patients with dengue with high viremia and increased severity of plasma leakage. Our findings therefore suggest that PCSK9 plays a hitherto unrecognized role in dengue pathogenesis and that PCSK9 inhibitors could be a suitable host-directed treatment for patients with dengue.
    MeSH term(s) Adolescent ; Adult ; Antiviral Agents/pharmacology ; Cell Hypoxia ; Cell Line ; Child ; Cholesterol/metabolism ; Dengue/drug therapy ; Dengue/etiology ; Dengue/metabolism ; Dengue Virus/pathogenicity ; Drug Resistance, Viral ; Female ; Hepatocytes/metabolism ; Hepatocytes/virology ; Humans ; Hydroxymethylglutaryl-CoA Reductase Inhibitors/pharmacology ; Male ; Myeloid Cells/metabolism ; Myeloid Cells/virology ; Proprotein Convertase 9/blood ; Proprotein Convertase 9/metabolism ; Receptors, LDL/metabolism ; Young Adult
    Chemical Substances Antiviral Agents ; Hydroxymethylglutaryl-CoA Reductase Inhibitors ; LDLR protein, human ; Receptors, LDL ; Cholesterol (97C5T2UQ7J) ; PCSK9 protein, human (EC 3.4.21.-) ; Proprotein Convertase 9 (EC 3.4.21.-)
    Language English
    Publishing date 2020-08-24
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 3067-3
    ISSN 1558-8238 ; 0021-9738
    ISSN (online) 1558-8238
    ISSN 0021-9738
    DOI 10.1172/JCI137536
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article: Visual and Biochemical Evidence of Glycocalyx Disruption in Human Dengue Infection, and Association With Plasma Leakage Severity.

    Lam, Phung Khanh / McBride, Angela / Le, Duyen Huynh Thi / Huynh, Trieu Trung / Vink, Hans / Wills, Bridget / Yacoub, Sophie

    Frontiers in medicine

    2020  Volume 7, Page(s) 545813

    Abstract: Background: ...

    Abstract Background:
    Language English
    Publishing date 2020-10-16
    Publishing country Switzerland
    Document type Journal Article
    ZDB-ID 2775999-4
    ISSN 2296-858X
    ISSN 2296-858X
    DOI 10.3389/fmed.2020.545813
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  3. Article ; Online: Using NS1 Flavivirus Protein Microarray to Infer Past Infecting Dengue Virus Serotype and Number of Past Dengue Virus Infections in Vietnamese Individuals.

    Thao, Tran Thi Nhu / de Bruin, Erwin / Phuong, Huynh Thi / Thao Vy, Nguyen Ha / van den Ham, Henk-Jan / Wills, Bridget A / Tien, Nguyen Thi Hanh / Le Duyen, Huynh Thi / Trung, Dinh The / Whitehead, Stephen S / Boni, Maciej F / Koopmans, Marion / Clapham, Hannah E

    The Journal of infectious diseases

    2020  Volume 223, Issue 12, Page(s) 2053–2061

    Abstract: Background: In recent years, researchers have had an increased focus on multiplex microarray assays, in which antibodies are measured against multiple related antigens, for use in seroepidemiological studies to infer past transmission.: Methods: We ... ...

    Abstract Background: In recent years, researchers have had an increased focus on multiplex microarray assays, in which antibodies are measured against multiple related antigens, for use in seroepidemiological studies to infer past transmission.
    Methods: We assess the performance of a flavivirus microarray assay for determining past dengue virus (DENV) infection history in a dengue-endemic setting, Vietnam. We tested the microarray on samples from 1 and 6 months postinfection from DENV-infected patients (infecting serotype was determined using reverse-transcription polymerase chain reaction during acute, past primary, and secondary infection assessed using plaque reduction neutralization tests 6 months postinfection).
    Results: Binomial models developed to discriminate past primary from secondary infection using the protein microarray (PMA) titers had high area under the curve (0.90-0.97) and accuracy (0.84-0.86). Multinomial models developed to identify most recent past infecting serotype using PMA titers performed well in those with past primary infection (average test set: κ = 0.85, accuracy of 0.92) but not those with past secondary infection (κ = 0.24, accuracy of 0.45).
    Conclusions: Our results suggest that the microarray will be useful in seroepidemiological studies aimed at classifying the past infection history of individuals (past primary vs secondary and serotype of past primary infections) and thus inferring past transmission intensity of DENV in dengue-endemic settings. Future work to validate these models should be undertaken in different transmission settings and with samples later after infection.
    MeSH term(s) Antibodies, Viral ; Asians ; Coinfection ; Dengue/epidemiology ; Dengue Virus/immunology ; Enzyme-Linked Immunosorbent Assay ; Flavivirus ; Humans ; Protein Array Analysis ; Serogroup ; Vietnam/epidemiology
    Chemical Substances Antibodies, Viral
    Language English
    Publishing date 2020-01-17
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 3019-3
    ISSN 1537-6613 ; 0022-1899
    ISSN (online) 1537-6613
    ISSN 0022-1899
    DOI 10.1093/infdis/jiaa018
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  4. Article ; Online: C-reactive protein as a potential biomarker for disease progression in dengue: a multi-country observational study.

    Vuong, Nguyen Lam / Le Duyen, Huynh Thi / Lam, Phung Khanh / Tam, Dong Thi Hoai / Vinh Chau, Nguyen Van / Van Kinh, Nguyen / Chanpheaktra, Ngoun / Lum, Lucy Chai See / Pleités, Ernesto / Jones, Nick Keith / Simmons, Cameron Paul / Rosenberger, Kerstin / Jaenisch, Thomas / Halleux, Christine / Olliaro, Piero Luigi / Wills, Bridget / Yacoub, Sophie

    BMC medicine

    2020  Volume 18, Issue 1, Page(s) 35

    Abstract: Background: Dengue infection can cause a wide spectrum of clinical outcomes. The severe clinical manifestations occur sufficiently late in the disease course, during day 4-6 of illness, to allow a window of opportunity for risk stratification. Markers ... ...

    Abstract Background: Dengue infection can cause a wide spectrum of clinical outcomes. The severe clinical manifestations occur sufficiently late in the disease course, during day 4-6 of illness, to allow a window of opportunity for risk stratification. Markers of inflammation may be useful biomarkers. We investigated the value of C-reactive protein (CRP) measured early on illness days 1-3 to predict dengue disease outcome and the difference in CRP levels between dengue and other febrile illnesses (OFI).
    Method: We performed a nested case-control study using the clinical data and samples collected from the IDAMS-consortium multi-country study. This was a prospective multi-center observational study that enrolled almost 8000 participants presenting with a dengue-like illness to outpatient facilities in 8 countries across Asia and Latin America. Predefined severity definitions of severe and intermediate dengue were used as the primary outcomes. A total of 281 cases with severe/intermediate dengue were compared to 836 uncomplicated dengue patients as controls (ratio 1:3), and also 394 patients with OFI.
    Results: In patients with confirmed dengue, median (interquartile range) of CRP level within the first 3 days was 30.2 mg/L (12.4-61.2 mg/L) (uncomplicated dengue, 28.6 (10.5-58.9); severe or intermediate dengue, 34.0 (17.4-71.8)). Higher CRP levels in the first 3 days of illness were associated with a higher risk of severe or intermediate outcome (OR 1.17, 95% CI 1.07-1.29), especially in children. Higher CRP levels, exceeding 30 mg/L, also associated with hospitalization (OR 1.37, 95% CI 1.14-1.64) and longer fever clearance time (HR 0.84, 95% CI 0.76-0.93), especially in adults. CRP levels in patients with dengue were higher than patients with potential viral infection but lower than patients with potential bacterial infection, resulting in a quadratic association between dengue diagnosis and CRP, with levels of approximately 30 mg/L associated with the highest risk of having dengue. CRP had a positive correlation with total white cell count and neutrophils and negative correlation with lymphocytes, but did not correlate with liver transaminases, albumin, or platelet nadir.
    Conclusions: In summary, CRP measured in the first 3 days of illness could be a useful biomarker for early dengue risk prediction and may assist differentiating dengue from other febrile illnesses.
    MeSH term(s) Adolescent ; Adult ; Biomarkers/blood ; C-Reactive Protein/analysis ; C-Reactive Protein/metabolism ; Case-Control Studies ; Child ; Child, Preschool ; Disease Progression ; Female ; Humans ; Male ; Middle Aged ; Prognosis ; Prospective Studies ; Severe Dengue/blood ; Severe Dengue/diagnosis ; Young Adult
    Chemical Substances Biomarkers ; C-Reactive Protein (9007-41-4)
    Language English
    Publishing date 2020-02-17
    Publishing country England
    Document type Journal Article ; Multicenter Study ; Research Support, Non-U.S. Gov't
    ISSN 1741-7015
    ISSN (online) 1741-7015
    DOI 10.1186/s12916-020-1496-1
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  5. Article ; Online: Endothelial Nitric Oxide Pathways in the Pathophysiology of Dengue: A Prospective Observational Study.

    Yacoub, Sophie / Lam, Phung Khanh / Huynh, Trieu Trung / Nguyen Ho, Hong Hanh / Dong Thi, Hoai Tam / Van, Nguyen Thu / Lien, Le Thi / Ha, Quyen Nguyen Than / Le, Duyen Huynh Thi / Mongkolspaya, Juthathip / Culshaw, Abigail / Yeo, Tsin Wen / Wertheim, Heiman / Simmons, Cameron / Screaton, Gavin / Wills, Bridget

    Clinical infectious diseases : an official publication of the Infectious Diseases Society of America

    2017  Volume 65, Issue 9, Page(s) 1453–1461

    Abstract: Background: Dengue can cause increased vascular permeability that may lead to hypovolemic shock. Endothelial dysfunction may underlie this; however, the association of endothelial nitric oxide (NO) pathways with disease severity is unknown.: Methods: ...

    Abstract Background: Dengue can cause increased vascular permeability that may lead to hypovolemic shock. Endothelial dysfunction may underlie this; however, the association of endothelial nitric oxide (NO) pathways with disease severity is unknown.
    Methods: We performed a prospective observational study in 2 Vietnamese hospitals, assessing patients presenting early (<72 hours of fever) and patients hospitalized with warning signs or severe dengue. The reactive hyperemic index (RHI), which measures endothelium-dependent vasodilation and is a surrogate marker of endothelial function and NO bioavailability, was evaluated using peripheral artery tonometry (EndoPAT), and plasma levels of l-arginine, arginase-1, and asymmetric dimethylarginine were measured at serial time-points. The main outcome of interest was plasma leakage severity.
    Results: Three hundred fourteen patients were enrolled; median age of the participants was 21(interquartile range, 13-30) years. No difference was found in the endothelial parameters between dengue and other febrile illness. Considering dengue patients, the RHI was significantly lower for patients with severe plasma leakage compared to those with no leakage (1.46 vs 2.00; P < .001), over acute time-points, apparent already in the early febrile phase (1.29 vs 1.75; P = .012). RHI correlated negatively with arginase-1 and positively with l-arginine (P = .001).
    Conclusions: Endothelial dysfunction/NO bioavailability is associated with worse plasma leakage, occurs early in dengue illness and correlates with hypoargininemia and high arginase-1 levels.
    MeSH term(s) Adolescent ; Adult ; Arginase/blood ; Arginase/metabolism ; Arginine/blood ; Arginine/metabolism ; Biomarkers/blood ; Biomarkers/metabolism ; Dengue/blood ; Dengue/epidemiology ; Dengue/metabolism ; Dengue/physiopathology ; Endothelium, Vascular/metabolism ; Female ; Humans ; Male ; Nitric Oxide/blood ; Nitric Oxide/metabolism ; Prospective Studies ; Young Adult
    Chemical Substances Biomarkers ; Nitric Oxide (31C4KY9ESH) ; Arginine (94ZLA3W45F) ; Arginase (EC 3.5.3.1)
    Language English
    Publishing date 2017-05-17
    Publishing country United States
    Document type Journal Article ; Observational Study
    ZDB-ID 1099781-7
    ISSN 1537-6591 ; 1058-4838
    ISSN (online) 1537-6591
    ISSN 1058-4838
    DOI 10.1093/cid/cix567
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  6. Article ; Online: Association of Microvascular Function and Endothelial Biomarkers With Clinical Outcome in Dengue: An Observational Study.

    Yacoub, Sophie / Lam, Phung Khanh / Vu, Le Hoang Mai / Le, Thi Lien / Ha, Ngo Thanh / Toan, Tran Thi / Van, Nguyen Thu / Quyen, Nguyen Than Ha / Le Duyen, Huynh Thi / Van Kinh, Nguyen / Fox, Annette / Mongkolspaya, Juthathip / Wolbers, Marcel / Simmons, Cameron Paul / Screaton, Gavin Robert / Wertheim, Heiman / Wills, Bridget

    The Journal of infectious diseases

    2016  Volume 214, Issue 5, Page(s) 697–706

    Abstract: Background: The hallmark of severe dengue is increased microvascular permeability, but alterations in the microcirculation and their evolution over the course of dengue are unknown.: Methods: We conducted a prospective observational study to evaluate ...

    Abstract Background: The hallmark of severe dengue is increased microvascular permeability, but alterations in the microcirculation and their evolution over the course of dengue are unknown.
    Methods: We conducted a prospective observational study to evaluate the sublingual microcirculation using side-stream dark-field imaging in patients presenting early (<72 hours after fever onset) and patients hospitalized with warning signs or severe dengue in Vietnam. Clinical findings, microvascular function, global hemodynamics assessed with echocardiography, and serological markers of endothelial activation were determined at 4 time points.
    Results: A total of 165 patients were enrolled. No difference was found between the microcirculatory parameters comparing dengue with other febrile illnesses. The proportion of perfused vessels (PPV) and the mean flow index (MFI) were lower in patients with dengue with plasma than those without leakage (PPV, 88.1% vs 90.6% [P = .01]; MFI, 2.1 vs 2.4 [P = .007]), most markedly during the critical phase. PPV and MFI were correlated with the endothelial activation markers vascular cell adhesion molecule 1 (P < .001 for both) and angiopoietin 2 (P < .001 for both), negatively correlated.
    Conclusions: Modest microcirculatory alterations occur in dengue, are associated with plasma leakage, and are correlate with molecules of endothelial activation, angiopoietin 2 and vascular cell adhesion molecule 1.
    MeSH term(s) Adolescent ; Adult ; Aged ; Aged, 80 and over ; Biomarkers/analysis ; Blood Vessels/diagnostic imaging ; Blood Vessels/pathology ; Capillary Permeability ; Child ; Dengue/diagnostic imaging ; Dengue/pathology ; Endothelial Cells/physiology ; Female ; Humans ; Male ; Middle Aged ; Optical Imaging ; Prospective Studies ; Treatment Outcome ; Vietnam ; Young Adult
    Chemical Substances Biomarkers
    Language English
    Publishing date 2016-05-26
    Publishing country United States
    Document type Journal Article ; Observational Study
    ZDB-ID 3019-3
    ISSN 1537-6613 ; 0022-1899
    ISSN (online) 1537-6613
    ISSN 0022-1899
    DOI 10.1093/infdis/jiw220
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