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Transcriptional regulation of IL-8 by Staphylococcus aureus in human conjunctival cells involves activation of AP-1.
Invest Ophthalmol Vis Sci. 2007 Jan; 48(1):270-6.IO

Abstract

PURPOSE

To identify signal transduction pathways involved in interleukin (IL)-8 expression by human conjunctival cells challenged with Staphylococcus aureus.

METHODS

Conjunctival cells were cultured in the presence of live or heat-killed S. aureus. IL-8 protein and mRNA were determined by ELISA and RT-PCR, respectively. Activation of mitogen-activated protein kinases (MAPKs) and NF-kappaB was analyzed by Western blot analysis with phosphospecific antibodies. Conjunctival cells were transfected with wild-type (wt) or mutated IL-8 promoters (IL-8-97, lacking the AP-1 site; IL-8-97 mutant C/EBP; IL-8-97 mutant NF-kappaB; IL-8/AP-1 double mutant for C/EBP and NF-kappaB) or c-Jun-NH(2)-terminal kinase (JNK)-responsive GAL-c-Jun. In further experiments, cells were cotransfected with wt IL-8 promoter and expression plasmids for p38MAPK-responsive C/EBP homologous protein (CHOP) or wt or dominant negative transactivation domain mutant (TAM-67) c-Jun. A protein-DNA binding study was performed by electrophoretic mobility shift assay (EMSA), to identify the transcription factors bound to the IL-8 promoter.

RESULTS

S. aureus induced significant IL-8 expression and synthesis in human conjunctival epithelial cells by activating c-Jun phosphorylation and transactivation potential via JNK. The IL-8 promoter activation was NF-kappaB- and p38MAPK-independent. Transfection and EMSA experiments suggested that only AP-1 transcription factors were necessary for optimal IL-8 expression.

CONCLUSIONS

Human conjunctival epithelial cells possess the ability to respond to Gram-positive S. aureus and to activate the innate immune response by the IL-8 gene expression. These results are the first to delineate the transcription factors involved in S. aureus-induced IL-8 release by conjunctival epithelium.

Authors+Show Affiliations

Departments of Surgical Specialties, The University of Messina, Messina, Italy.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

17197543

Citation

Venza, Isabella, et al. "Transcriptional Regulation of IL-8 By Staphylococcus Aureus in Human Conjunctival Cells Involves Activation of AP-1." Investigative Ophthalmology & Visual Science, vol. 48, no. 1, 2007, pp. 270-6.
Venza I, Cucinotta M, Caristi S, et al. Transcriptional regulation of IL-8 by Staphylococcus aureus in human conjunctival cells involves activation of AP-1. Invest Ophthalmol Vis Sci. 2007;48(1):270-6.
Venza, I., Cucinotta, M., Caristi, S., Mancuso, G., & Teti, D. (2007). Transcriptional regulation of IL-8 by Staphylococcus aureus in human conjunctival cells involves activation of AP-1. Investigative Ophthalmology & Visual Science, 48(1), 270-6.
Venza I, et al. Transcriptional Regulation of IL-8 By Staphylococcus Aureus in Human Conjunctival Cells Involves Activation of AP-1. Invest Ophthalmol Vis Sci. 2007;48(1):270-6. PubMed PMID: 17197543.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Transcriptional regulation of IL-8 by Staphylococcus aureus in human conjunctival cells involves activation of AP-1. AU - Venza,Isabella, AU - Cucinotta,Maria, AU - Caristi,Silvana, AU - Mancuso,Giuseppe, AU - Teti,Diana, PY - 2007/1/2/pubmed PY - 2007/2/6/medline PY - 2007/1/2/entrez SP - 270 EP - 6 JF - Investigative ophthalmology & visual science JO - Invest. Ophthalmol. Vis. Sci. VL - 48 IS - 1 N2 - PURPOSE: To identify signal transduction pathways involved in interleukin (IL)-8 expression by human conjunctival cells challenged with Staphylococcus aureus. METHODS: Conjunctival cells were cultured in the presence of live or heat-killed S. aureus. IL-8 protein and mRNA were determined by ELISA and RT-PCR, respectively. Activation of mitogen-activated protein kinases (MAPKs) and NF-kappaB was analyzed by Western blot analysis with phosphospecific antibodies. Conjunctival cells were transfected with wild-type (wt) or mutated IL-8 promoters (IL-8-97, lacking the AP-1 site; IL-8-97 mutant C/EBP; IL-8-97 mutant NF-kappaB; IL-8/AP-1 double mutant for C/EBP and NF-kappaB) or c-Jun-NH(2)-terminal kinase (JNK)-responsive GAL-c-Jun. In further experiments, cells were cotransfected with wt IL-8 promoter and expression plasmids for p38MAPK-responsive C/EBP homologous protein (CHOP) or wt or dominant negative transactivation domain mutant (TAM-67) c-Jun. A protein-DNA binding study was performed by electrophoretic mobility shift assay (EMSA), to identify the transcription factors bound to the IL-8 promoter. RESULTS: S. aureus induced significant IL-8 expression and synthesis in human conjunctival epithelial cells by activating c-Jun phosphorylation and transactivation potential via JNK. The IL-8 promoter activation was NF-kappaB- and p38MAPK-independent. Transfection and EMSA experiments suggested that only AP-1 transcription factors were necessary for optimal IL-8 expression. CONCLUSIONS: Human conjunctival epithelial cells possess the ability to respond to Gram-positive S. aureus and to activate the innate immune response by the IL-8 gene expression. These results are the first to delineate the transcription factors involved in S. aureus-induced IL-8 release by conjunctival epithelium. SN - 0146-0404 UR - https://www.unboundmedicine.com/medline/citation/17197543/Transcriptional_regulation_of_IL_8_by_Staphylococcus_aureus_in_human_conjunctival_cells_involves_activation_of_AP_1_ L2 - http://iovs.arvojournals.org/article.aspx?doi=10.1167/iovs.06-0081 DB - PRIME DP - Unbound Medicine ER -