title: Roles of cellular activation and sulfated glycans in Haemophilus somnus adherence to bovine brain microvascular endothelial cells.
abstract: Haemophilus somnus can cause a devastating fibrinopurulent meningitis with thrombotic vasculitis and encephalitis in cattle. The mechanisms used by H. somnus to migrate from the bloodstream into the central nervous system (CNS) are unknown. In this study, we demonstrate that H. somnus adheres to, but does not invade, bovine brain endothelial cells (BBEC) in vitro. The number of adherent H. somnus was significantly increased by prior activation of the BBEC with tumor necrosis factor alpha (TNF-alpha). Addition of exogenous glycosaminoglycans significantly reduced H. somnus adherence to resting and TNF-alpha-activated BBEC. Heparinase digestion of the endothelial cell's glycocalyx or sodium chlorate inhibition of endothelial cell sulfated glycan synthesis significantly reduced the number of adherent H. somnus. In contrast, addition of hyaluronic acid, a nonsulfated glycosaminoglycan, had no inhibitory effect. These findings suggest a critical role for both cellular activation and sulfated glycosaminoglycans in adherence of H. somnus to BBEC. Using heparin-labeled agarose beads, we demonstrated a high-molecular-weight heparin-binding protein expressed by H. somnus. Heparin was also shown to bind H. somnus in a 4 degrees C binding assay. These data suggest that heparin-binding proteins on H. somnus could serve as initial adhesins to sulfated proteoglycans on the endothelial cell surface, thus contributing to the ability of H. somnus to infect the bovine CNS.
authors.affiliation: Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin, 2015 Linden Drive, Madison, WI 53706, USA.
authors.forename: E
authors.initials: E
authors.lastname: Behling-Kelly
authors.forename: H
authors.initials: H
authors.lastname: Vonderheid
authors.forename: Kwang Sik
authors.initials: KS
authors.lastname: Kim
authors.forename: L B
authors.initials: LB
authors.lastname: Corbeil
authors.forename: C J
authors.initials: CJ
authors.lastname: Czuprynski
mesh_terms: D018829:Adhesins, Bacterial / Q000378:metabolism*; D000818:Animals; D001422:Bacterial Adhesion / Q000187:drug effects; D001812:Blood-Brain Barrier / Q000187:drug effects / Q000378:metabolism / Q000382:microbiology*; D001921:Brain / Q000382:microbiology / Q000473:pathology; D002417:Cattle / Q000382:microbiology*; D002704:Chlorates / Q000494:pharmacology; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism / Q000382:microbiology; D019276:Glycocalyx / Q000378:metabolism; D006025:Glycosaminoglycans / Q000494:pharmacology / Q000502:physiology*; D044138:Haemophilus somnus / Q000378:metabolism / Q000472:pathogenicity*; D006493:Heparin / Q000494:pharmacology; D006820:Hyaluronic Acid / Q000494:pharmacology; D011134:Polysaccharides / Q000494:pharmacology; D013431:Sulfates / Q000494:pharmacology; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
chemical_list: D018829:Adhesins, Bacterial; D002704:Chlorates; D006025:Glycosaminoglycans; D011134:Polysaccharides; D013431:Sulfates; D014409:Tumor Necrosis Factor-alpha; D006820:Hyaluronic Acid; D006493:Heparin; C032706:sodium chlorate