CORNEAL EPITHELIAL CELL SURFACE GLYCOCONJUGATES

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This two-part study will examine the pathogenesis of corneal infection and mechanisms of corneal epithelial cell migration. (1) Adherence of P. aeruginosa and S. Aureus to the Cornea: The goals of this study are to further elucidate the mechanism of bacterial attachment to corneal epithelium. The chemical structure of various rabbit corneal epithelial cell surface glycosphingolipids (GSL) which bind to P. aeruginosa and S. aureus will be elucidated. We will try to determine by histochemical techniques using anti-GSL monoclonal antibodies whether P. aeruginosa nd S. aureus-reactive GSL found in rabbit corneal epithelium in culture are (i) present on normal human and rabbit corneal epithelium in vivo, (ii) masked until injury causes them to be exposed, or (iii) present only on injured human and rabbit corneal epithelium. Experiments will also be performed to determine whether it is possible (i) to block the adherence of bacteria to the injured cornea by various anti-GSL monoclonal antibodies, sugars, oligosaccharides, lectins, and glycosidases and (ii) to elute the adherent bacteria from the injured corneas by various saccharide moieties. It is hoped that this study will contribute to our understanding of the pathogenesis of bacterial corneal ulcers. (2) Corneal Epithelial Wound Healing: The goal of this project are to clarify further the mechanism of corneal epithelial cell migration. The chemical structure of the migration-specific GSL of rabbit corneal epithelium will be elucidated. We will try to establish by histochemical techniques whether migration-specific GSL of rabbit corneas. It will also be determined whether it is possible to stimulate corneal epithelial migration of wounded corneas in organ culture by the addition of purified migration-specific GSL to the culture media. Experiments will also be performed to establish whether proteases play a role in corneal epithelial cell migration by exposing the masked corneal epitheal cell surface GSL and that these GSL influence cell migration by interacting with the macromolecules of the extracellular matrix. It is hoped that this study will contribute to our understanding of those factors that modulate corneal epithelial wound healing.