Tumour necrosis factor-alpha (TNF-alpha) increases nuclear factor kappaB (NFkappaB) activity in and interleukin-8 (IL-8) release from bovine mammary epithelial cells

D C Fitzgerald; K G Meade; A N McEvoy; L Lillis; E P Murphy; D E MacHugh; A W Baird

doi:10.1016/j.vetimm.2006.12.008

Tumour necrosis factor-alpha (TNF-alpha) increases nuclear factor kappaB (NFkappaB) activity in and interleukin-8 (IL-8) release from bovine mammary epithelial cells

D C Fitzgerald
, K G Meade
, A N McEvoy
, L Lillis
, E P Murphy
, D E MacHugh
, A W Baird

Research output: Contribution to journal › Article › peer-review

83 Citations (Scopus)

Abstract

Epithelia play important immunological roles at a variety of mucosal sites. We examined NFkappaB activity in control and TNF-alpha treated bovine mammary epithelial monolayers (BME-UV cells). A region of the bovine IL-8 (bIL-8) promoter was sequenced and a putative kappaB consensus sequence was identified bioinformatically. We used this sequence to analyse nuclear extracts for IL-8 specific NFkappaB activity. As a surrogate marker of NFkappaB 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-alpha enhanced NFkappaB activity in bovine mammary epithelial monolayers. p65 NFkappaB homodimer was identified in both control and TNF-alpha treated cells. Novel sequencing of the bovine IL-8 promoter identified a putative kappaB consensus sequence, which specifically bound TNF-alpha inducible p50/p65 heterodimer. TNF-alpha induced primarily serosal IL-8 release in the cell culture model. Pre-treatment with anti-TNF or dexamethasone inhibited TNF-alpha induced IL-8 release. High dose interleukin-1beta (IL-1beta) induced IL-8 release, however significantly less potently than TNF-alpha. Bovine mammary mucosal tissue released high basal levels of IL-8 which were unaffected by TNF-alpha or IL-1beta but inhibited by both dexamethasone and anti-TNF. These data support a role for TNF-alpha in activation of NFkappaB and release of IL-8 from bovine mammary epithelial cells.

Original language	English
Pages (from-to)	59-68
Number of pages	10
Journal	Veterinary Immunology and Immunopathology
Volume	116
Issue number	1-2
DOIs	https://doi.org/10.1016/j.vetimm.2006.12.008
Publication status	Published - 15 Mar 2007

Keywords

Animals
Anti-Inflammatory Agents
Antibodies, Monoclonal
Base Sequence
Cattle
Dexamethasone
Electrophoretic Mobility Shift Assay
Epithelial Cells
Female
Interleukin-8
Mammary Glands, Animal
Mastitis, Bovine
Molecular Sequence Data
Mucous Membrane
NF-kappa B
Tumor Necrosis Factor-alpha

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10.1016/j.vetimm.2006.12.008

Cite this

Fitzgerald, D. C., Meade, K. G., McEvoy, A. N., Lillis, L., Murphy, E. P., MacHugh, D. E., & Baird, A. W. (2007). Tumour necrosis factor-alpha (TNF-alpha) increases nuclear factor kappaB (NFkappaB) activity in and interleukin-8 (IL-8) release from bovine mammary epithelial cells. Veterinary Immunology and Immunopathology, 116(1-2), 59-68. https://doi.org/10.1016/j.vetimm.2006.12.008

@article{2ccd295a7c894256a8abfb5349b4ba0a,

title = "Tumour necrosis factor-alpha (TNF-alpha) increases nuclear factor kappaB (NFkappaB) activity in and interleukin-8 (IL-8) release from bovine mammary epithelial cells",

abstract = "Epithelia play important immunological roles at a variety of mucosal sites. We examined NFkappaB activity in control and TNF-alpha treated bovine mammary epithelial monolayers (BME-UV cells). A region of the bovine IL-8 (bIL-8) promoter was sequenced and a putative kappaB consensus sequence was identified bioinformatically. We used this sequence to analyse nuclear extracts for IL-8 specific NFkappaB activity. As a surrogate marker of NFkappaB 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-alpha enhanced NFkappaB activity in bovine mammary epithelial monolayers. p65 NFkappaB homodimer was identified in both control and TNF-alpha treated cells. Novel sequencing of the bovine IL-8 promoter identified a putative kappaB consensus sequence, which specifically bound TNF-alpha inducible p50/p65 heterodimer. TNF-alpha induced primarily serosal IL-8 release in the cell culture model. Pre-treatment with anti-TNF or dexamethasone inhibited TNF-alpha induced IL-8 release. High dose interleukin-1beta (IL-1beta) induced IL-8 release, however significantly less potently than TNF-alpha. Bovine mammary mucosal tissue released high basal levels of IL-8 which were unaffected by TNF-alpha or IL-1beta but inhibited by both dexamethasone and anti-TNF. These data support a role for TNF-alpha in activation of NFkappaB and release of IL-8 from bovine mammary epithelial cells.",

keywords = "Animals, Anti-Inflammatory Agents, Antibodies, Monoclonal, Base Sequence, Cattle, Dexamethasone, Electrophoretic Mobility Shift Assay, Epithelial Cells, Female, Interleukin-8, Mammary Glands, Animal, Mastitis, Bovine, Molecular Sequence Data, Mucous Membrane, NF-kappa B, Tumor Necrosis Factor-alpha",

author = "Fitzgerald, \{D C\} and Meade, \{K G\} and McEvoy, \{A N\} and L Lillis and Murphy, \{E P\} and MacHugh, \{D E\} and Baird, \{A W\}",

year = "2007",

month = mar,

day = "15",

doi = "10.1016/j.vetimm.2006.12.008",

language = "English",

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Fitzgerald, DC, Meade, KG, McEvoy, AN, Lillis, L, Murphy, EP, MacHugh, DE & Baird, AW 2007, 'Tumour necrosis factor-alpha (TNF-alpha) increases nuclear factor kappaB (NFkappaB) activity in and interleukin-8 (IL-8) release from bovine mammary epithelial cells', Veterinary Immunology and Immunopathology, vol. 116, no. 1-2, pp. 59-68. https://doi.org/10.1016/j.vetimm.2006.12.008

Tumour necrosis factor-alpha (TNF-alpha) increases nuclear factor kappaB (NFkappaB) activity in and interleukin-8 (IL-8) release from bovine mammary epithelial cells. / Fitzgerald, D C; Meade, K G; McEvoy, A N et al.
In: Veterinary Immunology and Immunopathology, Vol. 116, No. 1-2, 15.03.2007, p. 59-68.

Research output: Contribution to journal › Article › peer-review

TY - JOUR

T1 - Tumour necrosis factor-alpha (TNF-alpha) increases nuclear factor kappaB (NFkappaB) activity in and interleukin-8 (IL-8) release from bovine mammary epithelial cells

AU - Fitzgerald, D C

AU - Meade, K G

AU - McEvoy, A N

AU - Lillis, L

AU - Murphy, E P

AU - MacHugh, D E

AU - Baird, A W

PY - 2007/3/15

Y1 - 2007/3/15

N2 - Epithelia play important immunological roles at a variety of mucosal sites. We examined NFkappaB activity in control and TNF-alpha treated bovine mammary epithelial monolayers (BME-UV cells). A region of the bovine IL-8 (bIL-8) promoter was sequenced and a putative kappaB consensus sequence was identified bioinformatically. We used this sequence to analyse nuclear extracts for IL-8 specific NFkappaB activity. As a surrogate marker of NFkappaB 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-alpha enhanced NFkappaB activity in bovine mammary epithelial monolayers. p65 NFkappaB homodimer was identified in both control and TNF-alpha treated cells. Novel sequencing of the bovine IL-8 promoter identified a putative kappaB consensus sequence, which specifically bound TNF-alpha inducible p50/p65 heterodimer. TNF-alpha induced primarily serosal IL-8 release in the cell culture model. Pre-treatment with anti-TNF or dexamethasone inhibited TNF-alpha induced IL-8 release. High dose interleukin-1beta (IL-1beta) induced IL-8 release, however significantly less potently than TNF-alpha. Bovine mammary mucosal tissue released high basal levels of IL-8 which were unaffected by TNF-alpha or IL-1beta but inhibited by both dexamethasone and anti-TNF. These data support a role for TNF-alpha in activation of NFkappaB and release of IL-8 from bovine mammary epithelial cells.

AB - Epithelia play important immunological roles at a variety of mucosal sites. We examined NFkappaB activity in control and TNF-alpha treated bovine mammary epithelial monolayers (BME-UV cells). A region of the bovine IL-8 (bIL-8) promoter was sequenced and a putative kappaB consensus sequence was identified bioinformatically. We used this sequence to analyse nuclear extracts for IL-8 specific NFkappaB activity. As a surrogate marker of NFkappaB 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-alpha enhanced NFkappaB activity in bovine mammary epithelial monolayers. p65 NFkappaB homodimer was identified in both control and TNF-alpha treated cells. Novel sequencing of the bovine IL-8 promoter identified a putative kappaB consensus sequence, which specifically bound TNF-alpha inducible p50/p65 heterodimer. TNF-alpha induced primarily serosal IL-8 release in the cell culture model. Pre-treatment with anti-TNF or dexamethasone inhibited TNF-alpha induced IL-8 release. High dose interleukin-1beta (IL-1beta) induced IL-8 release, however significantly less potently than TNF-alpha. Bovine mammary mucosal tissue released high basal levels of IL-8 which were unaffected by TNF-alpha or IL-1beta but inhibited by both dexamethasone and anti-TNF. These data support a role for TNF-alpha in activation of NFkappaB and release of IL-8 from bovine mammary epithelial cells.

KW - Animals

KW - Anti-Inflammatory Agents

KW - Antibodies, Monoclonal

KW - Base Sequence

KW - Cattle

KW - Dexamethasone

KW - Electrophoretic Mobility Shift Assay

KW - Epithelial Cells

KW - Female

KW - Interleukin-8

KW - Mammary Glands, Animal

KW - Mastitis, Bovine

KW - Molecular Sequence Data

KW - Mucous Membrane

KW - NF-kappa B

KW - Tumor Necrosis Factor-alpha

U2 - 10.1016/j.vetimm.2006.12.008

DO - 10.1016/j.vetimm.2006.12.008

M3 - Article

C2 - 17276517

SN - 0165-2427

VL - 116

SP - 59

EP - 68

JO - Veterinary Immunology and Immunopathology

JF - Veterinary Immunology and Immunopathology

IS - 1-2

ER -

Tumour necrosis factor-alpha (TNF-alpha) increases nuclear factor kappaB (NFkappaB) activity in and interleukin-8 (IL-8) release from bovine mammary epithelial cells

Abstract

Keywords

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