title: Pro-inflammatory cytokine/chemokine gene expression in human corneal epithelial cells colonized by Pseudomonas aeruginosa.
abstract: Pro-inflammatory cytokines interleukin-6 (IL-6), interleukin-1beta (IL- 1beta), tumour necrosis factor-alpha (TNF-alpha) and chemokine interleukin-8 (IL-8) may play a significant role in the regulation of bacterial corneal infection. The aim of this study was to investigate the gene expression of these four mediators in a human corneal epithelial cell line challenged with Pseudomonas aeruginosa within 12 h. Human corneal epithelial monolayers were colonized with P. aeruginosa strains Paerl, 6206 and 6294. Expression of IL-6, IL-8, IL-1beta and TNF-alpha was analysed using semi-quantitative reverse transcription-polymerase chain reaction. Results showed that both IL-6 and IL-8 mRNA were expressed very early (4 h) during bacterial colonization and remained at high levels until the end of the experiment. Expression of IL-1beta and TNF-alpha mRNA appeared at 8 h after bacterial stimulation. No expression of IL-8, IL-1beta and TNF-alpha mRNA was observed in unstimulated cells. Interleukin-6 mRNA was expressed at low levels in unstimulated cells. In conclusion, bacterial colonization of human corneal epithelial cells induced expression of IL-6 and IL-8 mRNA earlier and at higher levels than IL-1beta and TNF-alpha mRNA.
authors.affiliation: Cooperative Research Centre for Eye Research and Technology, School of Optometry, University of New South Wales, Sydney, Australia. Mxue@cclru.randwick.unsw.edu.au
authors.forename: M L
authors.initials: ML
authors.lastname: Xue
authors.forename: A
authors.initials: A
authors.lastname: Thakur
authors.forename: L
authors.initials: L
authors.lastname: Lutze-Mann
authors.forename: M D
authors.initials: MD
authors.lastname: Willcox
mesh_terms: D002452:Cell Count; D002460:Cell Line; D018925:Chemokines / Q000096:biosynthesis / Q000235:genetics*; D015169:Colony Count, Microbial; D016207:Cytokines / Q000096:biosynthesis / Q000235:genetics*; D017931:DNA Primers; D018076:DNA, Complementary / Q000096:biosynthesis; D019573:Epithelium, Corneal / Q000378:metabolism / Q000382:microbiology*; D015870:Gene Expression; D006801:Humans; D011550:Pseudomonas aeruginosa / Q000502:physiology*; D012333:RNA, Messenger / Q000096:biosynthesis*; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D018925:Chemokines; D016207:Cytokines; D017931:DNA Primers; D018076:DNA, Complementary; D012333:RNA, Messenger