Effects of eosinophil granule proteins on human corneal epithelial cell viability and morphology.

Abstract

PURPOSE: There is mounting evidence that eosinophil granule proteins may cause tissue injury during allergic inflammation of the eye. Therefore, the authors investigated the in vitro effects of human eosinophil major basic protein (MBP), eosinophil cationic protein (ECP), eosinophil peroxidase (EPO), and eosinophil-derived neurotoxin (EDN) on cultured human corneal epithelial cell viability and morphology. METHODS: Confluent primary human corneal epithelial cell cultures were exposed to each of the four human eosinophil cationic granule proteins at concentrations ranging from 0 to 100 micrograms/ml (0, 12.5, 25, 50, and 100 micrograms/ml) for up to 48 hours in serum-free media. Morphologic changes were assessed by light microscopy at 1, 6, 24, and 48 hours; cell viability was measured using the MTT cell viability assay at 24 hours. RESULTS: Cells treated with MBP and ECP induced a dose-dependent gradual increase in morphologic changes; in contrast, EPO and EDN induced minimal changes in cell morphology. At 24 hours, both MBP and ECP induced statistically significant (P < 0.05) decreases in cell viability at a concentration of 100 micrograms/ml; EPO induced a significant (P < 0.05) decrease in cell viability at all concentrations tested, and EDN showed no significant reduction of cell viability at any of the concentrations tested. CONCLUSIONS: The current study suggests that the human eosinophil granule proteins MBP and ECP affect human corneal epithelial cell viability and morphology in vitro, whereas the protein EPO affects cell viability only. EDN had no significant effect on cell viability or morphology. Hence, MBP, ECP, and EPO perturb the corneal epithelium differentially and may contribute to keratopathy associated with severe ocular allergy.