Allergic eye disease, in some form, affects greater than 20 million people in the United States. To date, no systematic investigation has been undertaken to understand the molecular signaling events on the ocular surface of patients undergoing allergic reactions, or to explain the mechanism of cellular infiltration in allergic eye disease patients. The purpose of our research is to determine if activated human conjunctival mast cells will supply sufficient cytokines and other mediators to initiate and direct a well-orchestrated trafficking of eosinophils and other inflammatory cells to the ocular surface, facilitate their adhesion, and cause their release of potent effectors of ocular surface damage.

Second, we hope to determine the differences in molecular signaling in patients undergoing seasonal allergic conjunctivitis compared to those with sight threatening disease, such as atopic keratoconjunctivitis. We use methods that include activation with anti-IgE or secretagogue of human conjunctival mast cells. Evaluation of the levels of cytokines released is by ELISA, micro bead assay, or by examining the RNA by RT-PCR. Following treatment with activated supernatants from conjunctival mast cells, human conjunctival and corneal epithelial cells are then evaluated by FACS, RT-PCR, and ELISA for production of cell adhesion molecules and eosinophil-attracting chemokines. Eosinophil adhesion to these stimulated epithelial cells will be measured by eosinophil peroxidase adhesion assays. Eosinophils, following attachment to these epithelial cells, will be evaluated for activation by evaluation of oxidative burst, and release of potent effectors of epithelial cell damage, such as eosinophil cationic protein. These in-vitro results guide our in-vivo evaluation of ocular surface cells, which are obtained by impression cytology from patients undergoing an allergic reactions induced by topical provocation with allergen.

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Stahl JL, Cook EB, Barney NP, Graziano FM:  Mechanisms of antihistamines and mast cell stabilizers in ocular allergic inflammation.  Current Drug Targets – Inflammation and Allergy 2002,1:167-180.

Barney NP, Cook EB, Stahl JL, Graziano FM:  Pathophysiology of ocular allergy: the roles of conjunctival mast cells and epithelial cells.  Curr Allergy Asthma Rep 2002;2:332-339.

Stahl JL, Cook EB, Graziano FM, Barney NP.  Differential and cooperative effects of TNF alpha, IL-1 beta, and IFN gamma on human cell conjunctival epithelial cell receptor expression and chemokine release.  Invest Ophthalmol Visual Sci 2003;44(5):2010-2015.

Cook EB, Stahl JL, Sedgwick JB, Barney NP, Graziano FM:  The promotion of eosinophils degranulation and adhesion to conjunctival epithelial cells by IgE-activated conjunctival mast cells.  Ann Allergy Asthma Immunol 2004;92(1):65-72.