Document Detail


Role for 3-O-sulfated heparan sulfate as the receptor for herpes simplex virus type 1 entry into primary human corneal fibroblasts.
MedLine Citation:
PMID:  16940509     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Herpes simplex virus type 1 (HSV-1) infection of the corneal stroma remains a major cause of blindness. Primary cultures of corneal fibroblasts (CF) were tested and found susceptible to HSV-1 entry, which was confirmed by deconvolution imaging of infected cells. Plaque assay and real-time PCR demonstrated viral replication and hence a productive infection of CF by HSV-1. A role for glycoprotein D (gD) receptors in cultured CF was determined by gD interference assay. Reverse transcription-PCR analysis indicated expression of herpesvirus entry mediator and 3-O-sulfated (3-OS) heparan sulfate (HS)-generating enzyme 3-O sulfotransferase 3 (3-OST-3) but not nectin-1 or nectin-2. Subsequently, HS isolated from these cells was found to contain two distinct disaccharides (IdoUA2S-AnMan3S and IdoUA2S-AnMan3S6S) that are representative of 3-OST-3 activity. The following lines of evidence supported the important role of 3-OS HS as the mediator of HSV-1 entry into CF. (i) Blockage of entry was observed in CF treated with heparinases. The same enzymes had significantly less effect on HeLa cells that use nectin-1 as the entry receptor. (ii) Enzymatic removal of cell surface HS also removed the major gD-binding receptor, as evident from the reduced binding of gD to cells. (iii) Spinoculation assay demonstrated that entry blockage by heparinase treatment included the membrane fusion step. (iv) HSV-1 glycoprotein-induced cell-to-cell fusion was inhibited by either prior treatment of cells with heparinases or by HS preparations enriched in 3-OS HS. Taken together, the data in this report provide novel information on the role of 3-OS HS in mediating infection of CF, a natural target cell type.
Authors:
Vaibhav Tiwari; Christian Clement; Ding Xu; Tibor Valyi-Nagy; Beatrice Y J T Yue; Jian Liu; Deepak Shukla
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of virology     Volume:  80     ISSN:  0022-538X     ISO Abbreviation:  J. Virol.     Publication Date:  2006 Sep 
Date Detail:
Created Date:  2006-08-30     Completed Date:  2006-10-20     Revised Date:  2010-09-16    
Medline Journal Info:
Nlm Unique ID:  0113724     Medline TA:  J Virol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  8970-80     Citation Subset:  IM    
Affiliation:
University of Illinois at Chicago, Lions of Illinois Eye Research Institute, M/C 648, 1855 West Taylor Street, Chicago, IL 60612, USA.
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MeSH Terms
Descriptor/Qualifier:
Animals
CHO Cells
Cornea / cytology,  virology*
Cricetinae
Disaccharides / chemistry
Fibroblasts / cytology,  virology*
Glycoproteins / metabolism
Hela Cells
Heparin Lyase / metabolism
Heparitin Sulfate / chemistry*
Herpesvirus 1, Human / metabolism*
Humans
Protein Binding
Grant Support
ID/Acronym/Agency:
R01 AI 050050/AI/NIAID NIH HHS; R01 AI 057860/AI/NIAID NIH HHS; R01 EY 03890/EY/NEI NIH HHS
Chemical
Reg. No./Substance:
0/Disaccharides; 0/Glycoproteins; 9050-30-0/Heparitin Sulfate; EC 4.2.2.7/Heparin Lyase
Comments/Corrections

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