title: Cell surface ligands for rotavirus: mouse intestinal glycolipids and synthetic carbohydrate analogs.
abstract: Rotaviral binding to receptors on epithelial cells in the small intestine is thought to be a key event in the infection process and may be carbohydrate-mediated. Strain SA11 of rotavirus bound in vitro both to glycolipids isolated from mouse small intestine and to authentic glycolipids using thin layer chromatography overlay and microtiter well adsorption assays. Neutral mouse intestinal glycolipids which bound rotavirus were GA1 (Gal beta 1----3GalNAc beta 1---4Glc beta 1----4Glc beta 1----1-ceramide) and pentaosylceramides with terminal N-acetylgalactosamine, while acidic lipids which bound rotavirus included cholesterol 3-sulfate and two compounds termed bands 80 and 81. Digestion with ceramide glycanase suggested that bands 80 and 81 have lactosyl ceramide cores and an unidentified acidic moiety(s). No sialic-acid-containing glycolipids tested were active in viral binding. Band 81, which may have a ganglio core, bound rotavirus with greatest avidity, followed by GA1. Of authentic glycolipids assayed, only GA1 and GA2 (GalNAc beta 1----4Gal beta 1----4Glc beta 1----1-ceramide) displayed rotaviral binding. A phosphatidylethanolamide dipalmitoyl-containing neoglycolipid analog of GA2 bound rotavirus with avidity similar to native GA2. Substitution of beta 1----4-linked GlcNAc or beta 1----3-linked GalNAc for terminal GalNAc of GA2 neoglycolipid supported rotaviral binding, while other substitutions abrogated it. These findings suggest that a carbohydrate epitope similar to that of GA2 is sufficient for in vitro rotaviral binding, although binding may be enhanced by galactose and/or an acidic moiety in a secondary epitope.
authors.affiliation: Glycomed, Incorporated, Alameda, California 94501.
authors.forename: C A
authors.initials: CA
authors.lastname: Srnka
authors.forename: M
authors.initials: M
authors.lastname: Tiemeyer
authors.forename: J H
authors.initials: JH
authors.lastname: Gilbert
authors.forename: M
authors.initials: M
authors.lastname: Moreland
authors.forename: H
authors.initials: H
authors.lastname: Schweingruber
authors.forename: B W
authors.initials: BW
authors.lastname: de Lappe
authors.forename: P G
authors.initials: PG
authors.lastname: James
authors.forename: T
authors.initials: T
authors.lastname: Gant
authors.forename: R E
authors.initials: RE
authors.lastname: Willoughby
authors.forename: R H
authors.initials: RH
authors.lastname: Yolken
mesh_terms: D000818:Animals; D002240:Carbohydrate Sequence; D002845:Chromatography; D005677:G(M1) Ganglioside / Q000378:metabolism*; D005732:Gangliosides; D006017:Glycolipids / Q000378:metabolism*; D006028:Glycosphingolipids / Q000378:metabolism*; D007421:Intestine, Small / Q000737:chemistry* / Q000382:microbiology; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D011869:Radioligand Assay; D012401:Rotavirus / Q000378:metabolism*
chemical_list: D005732:Gangliosides; D006017:Glycolipids; D006028:Glycosphingolipids; C038244:ganglio-N-triaosylceramide; D005677:G(M1) Ganglioside; C018835:asialo GM1 ganglioside