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H Pereira, X Ruan, J Chodosh; Modulation of Adhesion Molecule Expression on Human Corneal Epithelial Cells (HCEC) by the Inflammatory Mediator, CAP37 . Invest. Ophthalmol. Vis. Sci. 2002;43(13):4201.
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Purpose: CAP37 an inflammatory mediator constitutively expressed in neutrophils (PMN) is an important host defense molecule. Recently, its expression was localized to corneal epithelial cells in an in vivo model of Staphylococcus aureus keratitis. The hypothesis proposed is that CAP37, either released by PMN or induced in corneal epithelial cells at the site of infection plays a pivotal role in the inflammatory response by modulating leukocyte and corneal epithelial cell functions, thereby regulating corneal inflammation and healing. Recent in vitro data from our laboratory indicates that CAP37 promotes corneal epithelial cell proliferation and migration, two important events in corneal wound healing. In this study we wished to pursue the role of CAP37 in regulating adhesion molecule expression in corneal epithelial cells, since attachment of newly formed epithelium to the extracellular matrix is essential for completing the healing process. We measured adhesion molecules ICAM-1, PECAM-1, VCAM-1 and E-selectin and alpha- and beta-integrins that are capable of binding to fibronectin, laminin, and other extracellular matrix proteins. Methods: Immortalized HCEC were treated with an optimal dose of CAP37 for 0-72 hr. Cell adhesion molecules ICAM-1, VCAM-1 PECAM-1 and E-selectin were measured by RT-PCR. Upregulation of integrin molecules on CAP37-treated HCEC were measured using flow cytometry. Results: CAP37 upregulated ICAM-1 and PECAM-1 in a time-dependent manner, with maximum upregulation observed at 4 hr. No upregulation of VCAM-1 or E-selectin mRNA was observed in response to CAP37. Flow cytometry indicated the upregulation of alpha-3 and beta-1 integrins, with peak upregulation between 12 and 24 hr. The integrin complex alpha-3beta-1 is important in mediating adhesion of corneal epithelial cells to corneal basement membrane laminins, 5 and 10. Conclusions: These findings demonstrate that CAP37 modulates corneal epithelial cell adhesion molecules involved in leukocyte-epithelial and epithelial-extracellular matrix interactions.
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