Lipopolysaccharide-induced Production of Interleukin-8 by Cultured Human Keratocyte

ABSTRACT

The authors performed an experiment to determine if human corneal keratocytes release IL-8 after stimulation with lipopolysaccharide (LPS). Human corneal keratocytes were isolated from human corneal buttoms and grown independently in vitro. Cultured keratocytes were treated with various concentrations of LPS (0.01, 0.1 1, 10 microgram/ml). At 6 hours, 12 hours, 24 hours, and 48 hours after the stimulation with LPS, culture supernatants were aspirated and frozen. Supernatants were assayed by enzyme-linked immunosorbent assay for IL-8 content. Exposure of corneal keratocytes to LPS induced IL-8 production. Initially, the secretion of IL-8 was detected at 6 hours and increased upto 48 hours. Between 12 and 24 hours, the IL-8 was increased rapidly. At 6 and 12 hours, keratocytes exposed to 10 microgram / ml LPS produced more IL-8 than those exposed to other concentrations of LPS. In this study, the ability of corneal keratocytes to produce IL-8 upto 48 hours suggests that these cells can play important roles in the induction of the inflammatory response in cornea.