Cell surface O-glycans limit Staphylococcus aureus adherence to corneal epithelial cells.
The mucin-rich environment of the intact corneal epithelium is thought to contribute to the prevention of Staphylococcus aureus infection. This study examined whether O-glycans, which constitute the majority of the mucin mass of epithelial cell glycocalyces, prevented bacterial adhesion and growth. Abrogation of mucin O glycosylation using the chemical primer benzyl-alpha-GalNAc resulted in increased adherence of parental strain RN6390 to apical human corneal-limbal epithelial (HCLE) cells and to biotinylated cell surface protein in static and liquid phase adhesion assays, consistent with a role of mucin O-glycans in preventing bacterial adhesion. Comparable results were found with ALC135, an isogenic mutant strain defective in the accessory gene regulators agr and sar, indicating that the agr- and/or sar-regulated virulence factors did not play a major role in mediating adhesion to the corneal cell surface after mucin O-glycan truncation. In exoglycosidase digestion studies, treatment with sialidase from Arthrobacter ureafaciens--which hydrolyzed mucin-associated O-acetyl sialic acid--but not from Clostridium perfringens resulted in an increase in RN6390 and ALC135 adhesion. Abrogation of mucin O glycosylation in HCLE cell cultures did not affect bacterial growth. Overall, these data indicate that mucin O-glycans contribute to the prevention of bacterial adherence to the apical surface of corneal epithelial cells and suggest that alteration of cell surface glycosylation from disease or trauma, including that stemming from contact lens wear, could contribute to a higher risk of infection.
Ricciuto
Jessica
J
Schepens Eye Research Institute and the Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts 02114, USA.
Heimer
Susan R
SR
Gilmore
Michael S
MS
Argüeso
Pablo
P
D000954:Antigens, Surface / Q000378:metabolism*; D001422:Bacterial Adhesion / Q000502:physiology*; D015153:Blotting, Western; D002478:Cells, Cultured; D019573:Epithelium, Corneal / Q000378:metabolism / Q000382:microbiology*; D006031:Glycosylation; D006801:Humans; D009077:Mucins / Q000378:metabolism; D019158:N-Acetylneuraminic Acid / Q000378:metabolism; D011134:Polysaccharides / Q000378:metabolism*; D013211:Staphylococcus aureus / Q000472:pathogenicity / Q000502:physiology*
D000954:Antigens, Surface; D009077:Mucins; C467440:O-acetyl sialic acid; D011134:Polysaccharides; D019158:N-Acetylneuraminic Acid
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