Abstract
Abstract: :
Purpose:The purpose of this study was to characterize the pattern of cytokine gene expression by human corneal epithelial cells (HCEC) in response to treatment with human interleukin-1, using a cytokine expression array. Methods:SV-40 transformed HCEC (SV40-HCEC) or primary cultured HCEC (P-HCEC) were treated for 6 hours with either serum-free growth-media alone or with the addition of human IL-1α or IL-1ß(10 ng/ml). Total RNA was extracted and 33-P labeled cDNA was generated by reverse transcription. A human cytokine expression array (R & D Systems, MN), that contained 375 different cytokines as cloned cDNA, was hybridized overnight with the radiolabeled cDNA. An autoradiograph was generated for each experimental condition and results were analyzed qualitatively. Results:IL-1α or IL-1ß treatment of HCEC produced similar induction of various cytokines. SV40-HCEC and P-HCEC demonstrated comparable cytokine profiles. IL-1 treatment modulated the expression of several genes. A comparison of the cytokine expression profiles revealed the following: Certain genes were turned ON only in the IL-1 treated group (e.g., Chemokines GRO ß, GRO γ; MCP-1; Proteases MMP-8, MMP-13); several genes were expressed in the media-treated cells but upregulated with IL-1 treatment (e.g., IL-8, IL-18; TNF-superfamily members TNF-α, TNF-R1, TRAIL R1, TRAIL R2, CD30, LT-ß; Proteases Furin, Urokinase; TGF-ß superfamily TGF-ßR1, Activins; Adhesion molecules P cadherin, R-cadherin); One gene - Integrin ß-2 - was turned OFF with IL-1 treatment while some genes were downregulated (e.g., Integrins αV, ß1) in the IL-1 treated group. Conclusion:IL-1 (α or ß) treatment of HCEC differentially regulates the expression of chemokines, other cytokines, cell-death related genes of the TNF-superfamily and cell-adhesion related genes. Such studies may provide a greater understanding of the ocular surface response in conditions of inflammation or corneal wound healing where the levels of IL-1 are known to be increased.
Keywords: 380 cytokines/chemokines • 370 cornea: basic science • 372 cornea: epithelium