This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Uchida, H.
Right arrow Articles by Glorioso, J. C.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Uchida, H.
Right arrow Articles by Glorioso, J. C.

 Previous Article  |  Next Article 

Journal of Virology, April 2009, p. 2951-2961, Vol. 83, No. 7
0022-538X/09/$08.00+0     doi:10.1128/JVI.01449-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Generation of Herpesvirus Entry Mediator (HVEM)-Restricted Herpes Simplex Virus Type 1 Mutant Viruses: Resistance of HVEM-Expressing Cells and Identification of Mutations That Rescue Nectin-1 Recognition{triangledown}

Hiroaki Uchida,1,2 Waris A. Shah,1,{dagger} Ali Ozuer,1 Arthur R. Frampton Jr.,1 William F. Goins,1 Paola Grandi,3 Justus B. Cohen,1 and Joseph C. Glorioso1*

Departments of Microbiology and Molecular Genetics,1 Surgery,2 Neurological Surgery, University of Pittsburgh, School of Medicine, Pittsburgh, Pennsylvania 152613

Received 10 July 2008/ Accepted 30 December 2008

Both initial infection and cell-to-cell spread by herpes simplex virus type 1 (HSV-1) require the interaction of the viral glycoprotein D (gD) with an entry receptor on the cell surface. The two major HSV entry receptors, herpesvirus entry mediator (HVEM) and nectin-1, mediate infection independently but are coexpressed on a variety of cells. To determine if both receptors are active in these instances, we have established mutant viruses that are selectively impaired for recognition of one or the other receptor. In plaque assays, these viruses showed approximately 1,000-fold selectivity for the matched receptor over the mismatched receptor. Separate assays showed that each virus is impaired for both infection and spread through the mismatched receptor. We tested several human tumor cell lines for susceptibility to these viruses and observed that HT29 colon carcinoma cells are susceptible to infection by nectin-1-restricted virus but are highly resistant to HVEM-restricted virus infection, despite readily detectable HVEM expression on the cell surface. HVEM cDNA isolated from HT29 cells rendered HSV-resistant cells permissive for infection by the HVEM-restricted virus, suggesting that HT29 cells lack a cofactor for HVEM-mediated infection or express an HVEM-specific inhibitory factor. Passaging of HVEM-restricted virus on nectin-1-expressing cells yielded a set of gD missense mutations that each restored functional recognition of nectin-1. These mutations identify residues that likely play a role in shaping the nectin-1 binding site of gD. Our findings illustrate the utility of these receptor-restricted viruses in studying the early events in HSV infection.

* Corresponding author. Mailing address: Department of Microbiology and Molecular Genetics, School of Medicine, University of Pittsburgh, E1240 BST, 200 Lothrop Street, Pittsburgh, PA 15261. Phone: (412) 648-8105. Fax: (412) 624-8997. E-mail: address: glorioso{at}pitt.edu

{triangledown} Published ahead of print on 7 January 2009.

{dagger} Present address: Centre for Research in Neuroscience, McGill University, Montreal General Hospital, L1-505, 1650 Cedar Avenue, Montreal H3G 1A4, Canada.

Journal of Virology, April 2009, p. 2951-2961, Vol. 83, No. 7
0022-538X/09/$08.00+0     doi:10.1128/JVI.01449-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»