Echoviruses bind heparan sulfate at the cell surface - PubMed (original) (raw)

Echoviruses bind heparan sulfate at the cell surface

I G Goodfellow et al. J Virol. 2001 May.

Abstract

Some echoviruses (EV) that bind decay-accelerating factor (DAF) also bind cells of human and murine origins in a DAF-independent manner. Pretreatment of cells with heparinase 1 or heparin blocks the binding of radiolabeled virus to the cell surface, and heparin prevents infection of rhabdomyosarcoma cells by certain EV, including several low-passage clinical isolates of EV 6 and some EV that do not bind DAF. These studies suggest that heparan sulfate may be of in vivo relevance as an attachment molecule for EV.

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Figures

FIG. 1

Involvement of GAGs in the DAF-independent binding of EV6 to rodent cells. 35S-Cys-Met metabolically labeled EV6 or EV7 (10,000 counts) was bound to the indicated cell lines for 2 h at 4°C, unbound virus was removed by washing with cold serum-free DMEM, and retained virus was quantified by scintillation counting. The mean and standard deviation for three independent experiments are shown.

FIG. 2

Heparin inhibits the binding (a) and infection (b) of RD cells by EV6. (a) Binding of radiolabeled EV6, clinical isolates of EV6 (indicated by the number symbol), EV7, EV9, and PV3 to RD cells was monitored in the presence of 1 mg of heparin/ml. Results are presented as the percentage of virus bound in comparison to binding of the untreated virus control (mean and standard deviation). (b) The inhibitory effect of heparin on infection of RD cells was determined by preincubating 1,000 TCID50 of the indicated viruses with dilutions of heparin (doubling dilutions from 2 mg/ml to 62.5 ng/ml) for 15 min before addition to 80% confluent RD cells. Infection was determined by staining with crystal violet after 24 h. The row labeled “Mock” indicates the effect of heparin in the absence of virus; lanes labeled 0 and De-N-Hep contained no heparin or 1 mg of de-_N_-sulfated heparin/ml, respectively.

FIG. 3

Relative effects of heparinase 1 and PIPLC on enterovirus binding to RD cells. Metabolically labeled, gradient-purified virus particles (10,000 counts) were bound to RD cells following pretreatment of the cell monolayer with heparinase 1 or PIPLC. The results presented are the mean and standard deviation for three independent assays and are expressed as the percentage bound in comparison to binding to untreated RD cells.

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Echoviruses bind heparan sulfate at the cell surface - PubMed (original) (raw)