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Journal of Virology, September 2001, p. 7818-7827, Vol. 75, No. 17
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.17.7818-7827.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Identification of a Putative Coreceptor on Vero Cells That Participates in Dengue 4 Virus Infection

José de Jesús Martínez-Barragán and Rosa M. del Angel*

Departamento de Patología Experimental, Centro de Investigación y de Estudios Avanzados del IPN, México City 07360, México

Received 1 February 2001/Accepted 29 May 2001

Dengue virus infects target cells by attaching to a cell surface receptor through the envelope (E) glycoprotein, located on the surface of the viral membrane. On Vero and BHK cells, heparan sulfate (HS) moieties of proteoglycans are the receptors for dengue virus; however, additional proteins have also been described as putative dengue virus receptors on C6/36, HL60, and BM cells. HS can also act as a receptor for other types of viruses or as an attachment molecule for viruses that require additional host cell molecules to allow viral penetration. In this study we searched for molecules other than HS that could participate in dengue virus infection of Vero cells. Labeled dengue 4 virus bound with high affinity to two molecules of 74 and 44 kDa. Binding of dengue virus to the 74-kDa molecule was susceptible to protease and sodium periodate treatment and resistant to heparinase treatments. Lectins such as concanavalin A and wheat germ agglutinin prevented dengue virus binding to both the 74- and the 44-kDa protein in overlay assays, while phytohemagglutinin P did not affect binding, suggesting that carbohydrate residues (alpha -mannose or N-acetylglucosamine) are important in virus binding to host cells. Protease susceptibility, biotin labeling, and immunofluorescence with a polyclonal antibody raised against the 74-kDa protein consistently identified the protein on the surfaces of Vero cells. Moreover, the antibody against the 74-kDa protein was able to inhibit dengue virus infection. These data suggest that HS might serve as a primary receptor, probably concentrating virus particles on the surfaces of Vero cells, and then other molecules, such as the 74-kDa protein, might participate as coreceptors in viral penetration. The 74-kDa protein possibly constitutes part of a putative receptor complex for dengue virus infection of Vero cells.

* Corresponding author. Mailing address: Departamento de Patología Experimental, Centro de Investigación y de Estudios Avanzados del IPN, Av. I.P.N. 2508. Col. San Pedro Zacatenco, México, D.F. C.P. 07360, México. Phone: (525) 747-7000, ext. 5648. Fax: (525) 747-7107. E-mail: rmangel{at}mail.cinvestav.mx.

Journal of Virology, September 2001, p. 7818-7827, Vol. 75, No. 17
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.17.7818-7827.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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