title: Production of interleukin-1 beta, interleukin-6 and tumor necrosis factor-alpha by a rat corneal epithelial cell line.
abstract: Production of interleukin (IL)-1 beta, IL-6 and tumor necrosis factor (TNF)-alpha by rat corneal epithelial cells in response to lipopolysaccharide and phorbol-12-myristate-13-acetate (PMA) was tested. Supernatants from rat corneal epithelial cells treated with lipopolysaccharide and PMA were collected after 6, 24 and 48 h and tested with enzyme-linked immunosorbent assay for IL-1 beta, IL-6 and TNF-alpha. The activity of TNF-alpha was additionally confirmed with bioassay on L929 cells. It was found that control groups did not produce significant levels of either cytokine. However, after stimulation with lipopolysaccharide, cells produced mainly IL-6, whereas after PMA they produced mainly TNF-alpha. IL-6 levels 24 and 48 h after PMA stimulation were also elevated, which could have been caused by the presence of TNF-alpha. Production of IL-1 beta in all groups was very low and remained within the test sensitivity range. These results show that the rat corneal epithelial cell line produces inflammatory cytokines in response to proinflammatory mediators. For this reason, it could be used for measuring the effects of irritants on the cornea.
authors.affiliation: Medical University of Warsaw, Department of Histology and Embryology, Warsaw, Poland. yaro@ib.amwaw.edu.pl
authors.forename: J
authors.initials: J
authors.lastname: Józwiak
authors.forename: P
authors.initials: P
authors.lastname: Skopiński
authors.forename: J
authors.initials: J
authors.lastname: Malejczyk
mesh_terms: D000826:Animal Testing Alternatives; D000818:Animals; D002273:Carcinogens / Q000494:pharmacology; D002460:Cell Line; D003315:Cornea / Q000166:cytology* / Q000276:immunology / Q000378:metabolism; D003358:Cosmetics; D004847:Epithelial Cells / Q000166:cytology / Q000276:immunology / Q000378:metabolism*; D007249:Inflammation / Q000139:chemically induced; D018836:Inflammation Mediators / Q000378:metabolism; D007375:Interleukin-1 / Q000096:biosynthesis*; D015850:Interleukin-6 / Q000096:biosynthesis*; D008070:Lipopolysaccharides / Q000494:pharmacology; D051381:Rats; D012680:Sensitivity and Specificity; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis*; D014945:Wound Healing / Q000187:drug effects / Q000276:immunology
chemical_list: D002273:Carcinogens; D003358:Cosmetics; D018836:Inflammation Mediators; D007375:Interleukin-1; D015850:Interleukin-6; D008070:Lipopolysaccharides; D014409:Tumor Necrosis Factor-alpha; D013755:Tetradecanoylphorbol Acetate