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Sulfated Escherichia coli K5 polysaccharide derivatives inhibit dengue virus infection of human microvascular endothelial cells by interacting with the viral envelope protein E domain III.

Vervaeke P, Alen M, Noppen S, Schols D, Oreste P, Liekens S - PLoS ONE (2013)

Bottom Line: Dengue virus (DENV) is an emerging mosquito-borne pathogen that causes cytokine-mediated alterations in the barrier function of the microvascular endothelium, leading to dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS).We observed that DENV (serotype 2) productively infects primary (HMVEC-d) and immortalized (HMEC-1) human dermal microvascular endothelial cells, despite the absence of well-described DENV receptors, such as dendritic cell-specific intercellular adhesion molecule-3-grabbing non-integrin (DC-SIGN) or the mannose receptor on the cell surface.Sulfated Escherichia coli K5 derivatives, which are structurally similar to heparin/heparan sulfate but lack anticoagulant activity, were able to block DENV infection of HMEC-1 and HMVEC-d cells in the nanomolar range.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology and Immunology, Rega Institute for Medical Research, KU Leuven, Leuven, Belgium.

ABSTRACT
Dengue virus (DENV) is an emerging mosquito-borne pathogen that causes cytokine-mediated alterations in the barrier function of the microvascular endothelium, leading to dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS). We observed that DENV (serotype 2) productively infects primary (HMVEC-d) and immortalized (HMEC-1) human dermal microvascular endothelial cells, despite the absence of well-described DENV receptors, such as dendritic cell-specific intercellular adhesion molecule-3-grabbing non-integrin (DC-SIGN) or the mannose receptor on the cell surface. However, heparan sulfate proteoglycans (HSPGs) were highly expressed on these cells and pre-treatment of HMEC-1 cells with heparinase II or with glycosaminoglycans reduced DENV infectivity up to 90%, suggesting that DENV uses HSPGs as attachment receptor on microvascular endothelial cells. Sulfated Escherichia coli K5 derivatives, which are structurally similar to heparin/heparan sulfate but lack anticoagulant activity, were able to block DENV infection of HMEC-1 and HMVEC-d cells in the nanomolar range. The highly sulfated K5-OS(H) and K5-N,OS(H) inhibited virus attachment and subsequent entry into microvascular endothelial cells by interacting with the viral envelope (E) protein, as shown by surface plasmon resonance (SPR) analysis using the receptor-binding domain III of the E protein.

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Anti-DENV-2 activity of sulfated GAGs.HMEC-1 cells were infected with DENV-2 at a MOI 1. Viral infectivity was quantified 24 h after infection by flow cytometry using an anti-DENV-2 specific antibody. Heparin (A), heparan sulfate (B), chondroitin sulfate A (C) and dermatan sulfate (D) dose-dependently inhibited DENV-2 infection in HMEC-1 cells. Data represent the % of infected cells relative to the positive control (DENV-2 infected cells). The means and standard deviations of three independent experiments are shown.
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pone-0074035-g003: Anti-DENV-2 activity of sulfated GAGs.HMEC-1 cells were infected with DENV-2 at a MOI 1. Viral infectivity was quantified 24 h after infection by flow cytometry using an anti-DENV-2 specific antibody. Heparin (A), heparan sulfate (B), chondroitin sulfate A (C) and dermatan sulfate (D) dose-dependently inhibited DENV-2 infection in HMEC-1 cells. Data represent the % of infected cells relative to the positive control (DENV-2 infected cells). The means and standard deviations of three independent experiments are shown.

Mentions: HSPG receptors are composed of a core protein and different GAG side-chains [60]. Therefore, the anti-DENV activity of naturally occurring GAGs (heparin, heparan sulfate, chondroitin sulfate A and dermatan sulfate) was investigated. A dose-dependent inhibitory activity was observed for all four GAGs by flow cytometry for viral antigen expression (Fig. 3). EC50 values were determined for each compound and are shown in Table 1. Heparin showed the most pronounced antiviral activity with an EC50 value (i.e. compound concentration that inhibits DENV infection by 50%) of 77 nM and could inhibit DENV-2 infection of HMEC-1 cells up to 93% at the highest concentration tested (7.7 µM). Heparan sulfate, chondroitin sulfate A and dermatan sulfate, in contrast, only showed antiviral activity (35–80%) at micromolar concentrations (3–6 µM).


Sulfated Escherichia coli K5 polysaccharide derivatives inhibit dengue virus infection of human microvascular endothelial cells by interacting with the viral envelope protein E domain III.

Vervaeke P, Alen M, Noppen S, Schols D, Oreste P, Liekens S - PLoS ONE (2013)

Anti-DENV-2 activity of sulfated GAGs.HMEC-1 cells were infected with DENV-2 at a MOI 1. Viral infectivity was quantified 24 h after infection by flow cytometry using an anti-DENV-2 specific antibody. Heparin (A), heparan sulfate (B), chondroitin sulfate A (C) and dermatan sulfate (D) dose-dependently inhibited DENV-2 infection in HMEC-1 cells. Data represent the % of infected cells relative to the positive control (DENV-2 infected cells). The means and standard deviations of three independent experiments are shown.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3755990&req=5

pone-0074035-g003: Anti-DENV-2 activity of sulfated GAGs.HMEC-1 cells were infected with DENV-2 at a MOI 1. Viral infectivity was quantified 24 h after infection by flow cytometry using an anti-DENV-2 specific antibody. Heparin (A), heparan sulfate (B), chondroitin sulfate A (C) and dermatan sulfate (D) dose-dependently inhibited DENV-2 infection in HMEC-1 cells. Data represent the % of infected cells relative to the positive control (DENV-2 infected cells). The means and standard deviations of three independent experiments are shown.
Mentions: HSPG receptors are composed of a core protein and different GAG side-chains [60]. Therefore, the anti-DENV activity of naturally occurring GAGs (heparin, heparan sulfate, chondroitin sulfate A and dermatan sulfate) was investigated. A dose-dependent inhibitory activity was observed for all four GAGs by flow cytometry for viral antigen expression (Fig. 3). EC50 values were determined for each compound and are shown in Table 1. Heparin showed the most pronounced antiviral activity with an EC50 value (i.e. compound concentration that inhibits DENV infection by 50%) of 77 nM and could inhibit DENV-2 infection of HMEC-1 cells up to 93% at the highest concentration tested (7.7 µM). Heparan sulfate, chondroitin sulfate A and dermatan sulfate, in contrast, only showed antiviral activity (35–80%) at micromolar concentrations (3–6 µM).

Bottom Line: Dengue virus (DENV) is an emerging mosquito-borne pathogen that causes cytokine-mediated alterations in the barrier function of the microvascular endothelium, leading to dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS).We observed that DENV (serotype 2) productively infects primary (HMVEC-d) and immortalized (HMEC-1) human dermal microvascular endothelial cells, despite the absence of well-described DENV receptors, such as dendritic cell-specific intercellular adhesion molecule-3-grabbing non-integrin (DC-SIGN) or the mannose receptor on the cell surface.Sulfated Escherichia coli K5 derivatives, which are structurally similar to heparin/heparan sulfate but lack anticoagulant activity, were able to block DENV infection of HMEC-1 and HMVEC-d cells in the nanomolar range.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology and Immunology, Rega Institute for Medical Research, KU Leuven, Leuven, Belgium.

ABSTRACT
Dengue virus (DENV) is an emerging mosquito-borne pathogen that causes cytokine-mediated alterations in the barrier function of the microvascular endothelium, leading to dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS). We observed that DENV (serotype 2) productively infects primary (HMVEC-d) and immortalized (HMEC-1) human dermal microvascular endothelial cells, despite the absence of well-described DENV receptors, such as dendritic cell-specific intercellular adhesion molecule-3-grabbing non-integrin (DC-SIGN) or the mannose receptor on the cell surface. However, heparan sulfate proteoglycans (HSPGs) were highly expressed on these cells and pre-treatment of HMEC-1 cells with heparinase II or with glycosaminoglycans reduced DENV infectivity up to 90%, suggesting that DENV uses HSPGs as attachment receptor on microvascular endothelial cells. Sulfated Escherichia coli K5 derivatives, which are structurally similar to heparin/heparan sulfate but lack anticoagulant activity, were able to block DENV infection of HMEC-1 and HMVEC-d cells in the nanomolar range. The highly sulfated K5-OS(H) and K5-N,OS(H) inhibited virus attachment and subsequent entry into microvascular endothelial cells by interacting with the viral envelope (E) protein, as shown by surface plasmon resonance (SPR) analysis using the receptor-binding domain III of the E protein.

Show MeSH
Related in: MedlinePlus