Glycosaminoglycan Sulfation Requirements for Respiratory Syncytial Virus Infection

Hallak, Louay K.; Spillmann, Dorothe; Collins, Peter L.; Peeples, Mark E.

doi:10.1128/jvi.74.22.10508-10513.2000

Cited by 271 publications

(255 citation statements)

References 45 publications

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“…To address whether localization of the hRSV N at the cell surface requires active viral replication, we used a recombinant hRSV A2 strain encoding the green fluorescent protein (hRSV GFP ). Because GFP translation from the viral genome is regulated by the hRSV large RNA-dependent RNA polymerase, GFP synthesis serves as reporter for viral replication (17,30). Also, because the GFP displays a half-life of 26 h (31), accumulation of GFP over time [and the increase in its mean fluorescence intensity (MFI)] serves to track the progression of the viral replication cycle in infected cells.…”

Section: Resultsmentioning

confidence: 99%

Surface expression of the hRSV nucleoprotein impairs immunological synapse formation with T cells

Céspedes

Bueno

Ramírez

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

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Human respiratory syncytial virus (hRSV) is the leading cause of bronchiolitis and pneumonia in young children worldwide. The recurrent hRSV outbreaks and reinfections are the cause of a significant public health burden and associate with an inefficient antiviral immunity, even after disease resolution. Although several mouse-and human cell-based studies have shown that hRSV infection prevents naïve T-cell activation by antigen-presenting cells, the mechanism underlying such inhibition remains unknown. Here, we show that the hRSV nucleoprotein (N) could be at least partially responsible for inhibiting T-cell activation during infection by this virus. Early after infection, the N protein was expressed on the surface of epithelial and dendritic cells, after interacting with trans-Golgi and lysosomal compartments. Further, experiments on supported lipid bilayers loaded with peptide-MHC (pMHC) complexes showed that surface-anchored N protein prevented immunological synapse assembly by naive CD4 + T cells and, to a lesser extent, by antigen-experienced T-cell blasts. Synapse assembly inhibition was in part due to reduced T-cell receptor (TCR) signaling and pMHC clustering at the T-cell− bilayer interface, suggesting that N protein interferes with pMHC− TCR interactions. Moreover, N protein colocalized with the TCR independently of pMHC, consistent with a possible interaction with TCR complex components. Based on these data, we conclude that hRSV N protein expression at the surface of infected cells inhibits T-cell activation. Our study defines this protein as a major virulence factor that contributes to impairing acquired immunity and enhances susceptibility to reinfection by hRSV.

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Section: Resultsmentioning

confidence: 99%

Surface expression of the hRSV nucleoprotein impairs immunological synapse formation with T cells

Céspedes

Bueno

Ramírez

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

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“…The RSV attachment glycoprotein has been found to be dispensable for infection of cultured cells by RSV (58 -60); syncytium formation occurs with cells expressing the RSV fusion glycoprotein alone (61,62), and the RSV fusion glycoprotein appears to be able to mediate attachment via glycosaminoglycan interaction (26,58,60,63). However, recombinant HRSV passages 10 -20 times more efficiently in cell culture when both the G and fusion glycoproteins are present compared with when only the fusion glycoprotein is expressed (59).…”

Section: Discussionmentioning

confidence: 99%

“…The mechanism by which this mucin-like extramembranous domain mediates attachment of RSV to host cells or between infected and noninfected host cells is unknown (22). Glycosaminoglycans of cellular proteoglycans have been implicated in RSV attachment (23)(24)(25)(26)(27), but the identities of these proteoglycans or of other cellular receptors are unknown (22,27).…”

mentioning

confidence: 99%

Antiviral Activity and Structural Characteristics of the Nonglycosylated Central Subdomain of Human Respiratory Syncytial Virus Attachment (G) Glycoprotein

Gorman¹,

McKimm-Breschkin²,

Norton³

et al. 2001

Journal of Biological Chemistry

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“…The binding of RSV to host cells is facilitated by the presence of cellular glycosaminoglycans (GAGs), particularly heparan sulfate or other iduronic-acid-containing sugar chains (21,29,34). Feldman et al showed that both G and F can bind to immobilized heparin and that soluble GAGs can inhibit the binding and replication of both wild-type RSV and the coldpassaged isolate cp-52, which lacks the G and SH glycoproteins (16).…”

mentioning

confidence: 99%

RhoA-Derived Peptide Dimers Share Mechanistic Properties with Other Polyanionic Inhibitors of Respiratory Syncytial Virus (RSV), Including Disruption of Viral Attachment and Dependence on RSV G

Budge

Beeler

et al. 2004

J Virol

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Large polyanionic molecules, such as sulfated polysaccharides (including soluble heparin and dextran sulfate), synthetic polyanionic polymers, and negatively charged proteins, have been shown to broadly inhibit several enveloped viruses. We recently reported the antiviral activity of a peptide derived from amino acids 77 to 95 of a potential binding partner of respiratory syncytial virus F protein (RSV F), the GTPase RhoA. A subsequent study with a truncated peptide (amino acids 80 to 94) revealed that optimal antiviral activity required dimerization via intermolecular disulfide bonds. We report here that the net negative charge of this peptide is also a determining factor for its antiviral activity and that it, like other polyanions, inhibits virus attachment. In a flow cytometry-based binding assay, peptide 80-94, heparin, and dextran sulfate inhibited the attachment of virus to cells at 4°C at the same effective concentrations at which they prevent viral infectivity. Interestingly, time-of-addition experiments revealed that peptide 80-94 and soluble heparin were also able to inhibit the infectivity of a virus that had been prebound to cells at 4°C, as had previously been shown for dextran sulfate, suggesting a potential role for postattachment effects of polyanions on RSV entry. Neutralization experiments with recombinant viruses showed that the antiviral activities of peptide 80-94 and dextran sulfate were diminished in the absence of the RSV attachment glycoprotein (G). Taken together, these data indicate that the antiviral activity of RhoA-derived peptides is functionally similar to that of other polyanions, is dependent on RSV G, and does not specifically relate to a protein-protein interaction between F and RhoA.

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Glycosaminoglycan Sulfation Requirements for Respiratory Syncytial Virus Infection

Abstract: Glycosaminoglycans (GAGs

Cited by 271 publications

References 45 publications

Surface expression of the hRSV nucleoprotein impairs immunological synapse formation with T cells

Surface expression of the hRSV nucleoprotein impairs immunological synapse formation with T cells

Antiviral Activity and Structural Characteristics of the Nonglycosylated Central Subdomain of Human Respiratory Syncytial Virus Attachment (G) Glycoprotein

RhoA-Derived Peptide Dimers Share Mechanistic Properties with Other Polyanionic Inhibitors of Respiratory Syncytial Virus (RSV), Including Disruption of Viral Attachment and Dependence on RSV G

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