ABSTRACT
PURPOSE: The interferon-gamma-inducing factor Interleukin-18 (IL-18) is a recently described cytokine that appears to have multiple important pro-inflammatory effects including the induction of interferon-gamma (IFN-gamma) by activated T-cells. The expression of IL-18 by human cornea has not been previously reported. In the present study, we examine the possibility that human corneal epithelial cells are capable of producing this leukocyte-activating factor which may play an important role in IFN-gamma-dependent inflammation responses in the cornea. METHODS: Northern blot analysis and ELISA were used to investigate the in vitro expression of IL-18 mRNA and protein respectively in primary (HCEC) and transformed human corneal epithelial cells (HCET). To determine if IL-18 expression was modulated by pro-inflammatory mediators, cells were treated with lipopolysaccharide (LPS), phorbol 12-myristate 13-acetate (PMA) or synthetic double stranded RNA (poly dI : dC). IL-18 bioactivity was determined in a leukocyte interferon-gamma induction assay and IL-18 was immunolocalized in whole human cornea by immunohistochemistry using a specific anti-IL-18 antibody. RESULTS: IL-18 mRNA and bioactive protein was constitutively expressed by human corneal epithelial cells and upregulated by PMA, LPS and poly dI : dC. The constitutive expression of IL-18 protein immunoreactivity was also demonstrated in the epithelial cells of whole human cornea tissue. CONCLUSIONS: This is the first study demonstrating that corneal epithelial cells are capable of producing the IFN-gamma inducing factor IL-18. Increased bioactive corneal IL-18 production can be induced by a number of pro-inflammatory agents and may play an important role in initiating gamma-interferon-mediated inflammatory responses in the cornea.
Subject(s)
Epithelium, Corneal/metabolism , Interleukin-18/biosynthesis , Blotting, Northern , Cell Line , Enzyme-Linked Immunosorbent Assay , Epithelium, Corneal/drug effects , Gene Expression Regulation , Humans , Immunoenzyme Techniques , Interleukin-18/genetics , Lipopolysaccharides/pharmacology , Polydeoxyribonucleotides/pharmacology , RNA, Messenger/biosynthesis , Tetradecanoylphorbol Acetate/pharmacologyABSTRACT
The activities of 10 enzymes involved in carbohydrate metabolism were measured in both desiccated and rehydrated fronds of the desiccation-tolerant pteridophyte Selaginella lepidophylla (Hook. & Grev.) Spring. Enzyme conservation was defined as the ratio of desiccated to hydrated frond enzyme activity. The mean level of conservation was 74%, with nine of the 10 enzymes showing significant activity increases (P<0.05) during hydration. The mean of photosynthetic enzyme conservation was significantly lower (P=0.05) than the mean for glycolytic and respiratory enzymes combined. Chloramphenicol inhibited the normal activity increase in ribulose bisphosphate carboxylase and (NADPH)triose-P dehydrogenase but not pyruvate kinase upon rehydration. Cycloheximide did not affect the normal activity increase for these three enzymes. It is concluded that substantial enzyme conservation is beneficial for rapid resumption of metabolic activity in resurrection plants.
