Autor: |
L. Lillis, David E. MacHugh, Evelyn P. Murphy, Alice N. McEvoy, Kieran G. Meade, Alan W. Baird, Denise C. Fitzgerald |
Rok vydání: |
2007 |
Předmět: |
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Zdroj: |
Veterinary Immunology and Immunopathology. 116:59-68 |
ISSN: |
0165-2427 |
DOI: |
10.1016/j.vetimm.2006.12.008 |
Popis: |
Epithelia play important immunological roles at a variety of mucosal sites. We examined NFκB activity in control and TNF-α treated bovine mammary epithelial monolayers (BME-UV cells). A region of the bovine IL-8 (bIL-8) promoter was sequenced and a putative κB consensus sequence was identified bioinformatically. We used this sequence to analyse nuclear extracts for IL-8 specific NFκB activity. As a surrogate marker of NFκB activation, we investigated IL-8 release in two models. Firstly in BME-UV monolayers, IL-8 release in the presence of pro- and anti-inflammatory agents was determined by enzyme-linked immunosorbent assay (ELISA). Secondly, we measured IL-8 secretion from a novel model of intact mucosal sheets of bovine teat sinus. IL-8 release into bathing solutions was assessed following treatment with pro- and anti-inflammatory agents. TNF-α enhanced NFκB activity in bovine mammary epithelial monolayers. p65 NFκB homodimer was identified in both control and TNF-α treated cells. Novel sequencing of the bovine IL-8 promoter identified a putative κB consensus sequence, which specifically bound TNF-α inducible p50/p65 heterodimer. TNF-α induced primarily serosal IL-8 release in the cell culture model. Pre-treatment with anti-TNF or dexamethasone inhibited TNF-α induced IL-8 release. High dose interleukin-1β (IL-1β) induced IL-8 release, however significantly less potently than TNF-α. Bovine mammary mucosal tissue released high basal levels of IL-8 which were unaffected by TNF-α or IL-1β but inhibited by both dexamethasone and anti-TNF. These data support a role for TNF-α in activation of NFκB and release of IL-8 from bovine mammary epithelial cells. |
Databáze: |
OpenAIRE |
Externí odkaz: |
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