Abstract

Background: Allergic eye disease, in some form, affects greater than 20 million people in the US. No systematic investigation has been undertaken to attempt to understand the molecular signaling events on the ocular surface and to explain the mechanism and course of cellular infiltration in allergic eye disease patients. Purpose: 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 to the ocular surface, facilitate their adhesion, and cause their release of potent affecters of ocular surface damage. Secondly, to determine the differences in molecular signaling in patients undergoing seasonal allergic conjunctivitis compared to those with sight threatening disease such as Atopic Keratoconjunctivitis. Methods: Following activation with anti-IgE or secretagogue, human conjunctival mast cell release of cytokines will be evaluated by ELISA, RT-PCR, or micro bead assay. Following treatment with activated supernatants from conjunctival mast cells (identifying important cytokines using blocking antibodies), human conjunctival or corneal epithelial cells, will be 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. Following their attachment to epithelial cells, eosinophils will be measured for activation by evaluation of oxidative burst and for release of potent affecters of epithelial cell damage such as eosinophil cationic protein by ELISA. These in-vitro results will guide our in vivo evaluation (FACS, RT-PCR) of ocular surface cells (obtained by impression etiology) of patients undergoing an allergic reaction induced by topical provocation with allergen. Conclusion: We will be able to connect the molecular signaling events of cells of the ocular surface to the known pathologic findings of allergic eye diseases. This understanding will allow the development of precise strategies for intervention.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
5R01EY012526-02
Application #
6179074
Study Section
Visual Sciences A Study Section (VISA)
Program Officer
Fisher, Richard S
Project Start
1999-04-01
Project End
2002-03-31
Budget Start
2000-04-01
Budget End
2001-03-31
Support Year
2
Fiscal Year
2000
Total Cost
$211,203
Indirect Cost

  Institution

Name
University of Wisconsin Madison
Department
Ophthalmology
Type
Schools of Medicine
DUNS #
161202122
City
Madison
State
WI
Country
United States
Zip Code
53715

  Publications

Nansera, D; Graziano, F M; Friedman, D J et al. (2011) Vitamin D and calcium levels in Ugandan adults with human immunodeficiency virus and tuberculosis. Int J Tuberc Lung Dis 15:1522-7, i
Nansera, D; Bajunirwe, F; Kabakyenga, J et al. (2010) Opportunities and barriers for implementation of integrated TB and HIV care in lower level health units: experiences from a rural western Ugandan district. Afr Health Sci 10:312-9
Cook, Ellen B; Stahl, James L; Graziano, Frank M et al. (2008) Regulation of the receptor for TNFalpha, TNFR1, in human conjunctival epithelial cells. Invest Ophthalmol Vis Sci 49:3992-8
Cook, Ellen B; Stahl, James L; Brooks, Anne M et al. (2006) Allergic tears promote upregulation of eosinophil adhesion to conjunctival epithelial cells in an ex vivo model: inhibition with olopatadine treatment. Invest Ophthalmol Vis Sci 47:3423-9
Cook, Ellen B; Stahl, James L; Sedgwick, Julie B et al. (2004) The promotion of eosinophil degranulation and adhesion to conjunctival epithelial cells by IgE-activated conjunctival mast cells. Ann Allergy Asthma Immunol 92:65-72
Stahl, James L; Cook, Ellen B; Graziano, Frank M et al. (2003) Differential and cooperative effects of TNFalpha, IL-1beta, and IFNgamma on human conjunctival epithelial cell receptor expression and chemokine release. Invest Ophthalmol Vis Sci 44:2010-5
Cook, E B; Stahl, J L; Barney, N P et al. (2002) Mechanisms of antihistamines and mast cell stabilizers in ocular allergic inflammation. Curr Drug Targets Inflamm Allergy 1:167-80
Stahl, James L; Cook, Ellen B; Barney, Neal P et al. (2002) Pathophysiology of ocular allergy: the roles of conjunctival mast cells and epithelial cells. Curr Allergy Asthma Rep 2:332-9
Lee, Catherine H; Whiteman, Amanda L; Murphy, Christopher J et al. (2002) Substance P, insulinlike growth factor 1, and surface healing. Arch Ophthalmol 120:215-7
Cook, E B; Stahl, J L; Barney, N P et al. (2001) Olopatadine inhibits anti-immunoglobulin E-stimulated conjunctival mast cell upregulation of ICAM-1 expression on conjunctival epithelial cells. Ann Allergy Asthma Immunol 87:424-9

Showing the most recent 10 out of 12 publications