Interaction Between Respiratory Syncytial Virus and Glycosaminoglycans, Including Heparan Sulfate

Abstract

Glycosaminoglycans (GAGs), including heparan sulfate (HS), are expressed on the surface of nearly all cells, linked to transmembrane proteins. These GAGs are sulfated to varying extents, lending a negative charge, and are used by a large number of viruses to initiate infection of immortalized cell lines. Here we describe the rationale and methods for analyzing GAG usage by one such virus, respiratory syncytial virus (RSV). The protocols presented allow the determination of which GAG(s) is employed by the virus, which GAG modification(s) is important, and whether the important GAG is on the cell or on the virus. We also discuss the finding that many viruses are selected for GAG usage during passage in culture and present a method for rapidly determining whether GAG usage is characteristic of a wild virus or is limited to laboratory-adapted virus.

Recombinant Subgroup B Human Respiratory Syncytial Virus Expressing Enhanced Green Fluorescent Protein Efficiently Replicates in Primary Human Cells and Is Virulent in Cotton Rats

“…1J). It has been reported that HRSV strains differ in their binding affinity to glycosaminoglycan (GAG) moieties on the cell surface (19,23). Both rHRSV B05 and rHRSV B05 EGFP(5) displayed low GAG dependency indices (Fig.…”

“…Titers are expressed as 50% tissue culture infectious doses (TCID 50 ) calculated by the Reed and Muench method (22). To determine glycosaminoglycan (GAG) indices, CHO cells expressing GAG or GAG-deficient cells were infected and analyzed by flow cytometry (23). wd-NHBE cells were cultured in 12-mm/0.4-m-pore-size inserts (Corning) at ALI (24).…”

Structure and Function of Respiratory Syncytial Virus Surface Glycoproteins

“…The receptor for the RSV G protein on immortalized cells appears to be HS (Feldman et al 1999; Feldman et al 2000; Hallak et al 2000a; Hallak et al 2000b; Krusat and Streckert 1997; Escribano-Romero et al 2004), similar to a number of other viruses (Hallak et al 2007). But HS is not detectable on the apical surface of HAE cultures (Zhang et al 2005) suggesting that RSV most likely uses a different receptor to enter these cells.…”

Respiratory Syncytial Virus Infection Upregulates NLRC5 and Major Histocompatibility Complex Class I Expression through RIG-I Induction in Airway Epithelial Cells

“…In patients, however, infection is normally highly restricted to the superficial cells of the respiratory epithelium, the ciliated cells of the small bronchioles, and pneumocytes in the alveoli (8)(9)(10). Infection is initiated by cell surface binding via proteoglycans (11), followed by nucleolin-mediated fusion for RSV cell entry (9,12) and infection. In response, the host initiates an early innate immune response at the site of infection.…”