NF-κB activation is required for C5a-induced interleukin-8 gene expression in mononuclear cells

Matthew H. Hsu, Meiying Wang, Darren D Browning, Naofumi Mukaida, Richard D. Ye

Research output: Contribution to journalArticle

43 Citations (Scopus)

Abstract

C5a, a potent peptide chemoattractant, stimulates interleukin-8 (IL-8) secretion from peripheral blood mononuclear cells (PBMC). Experiments were conducted to understand the mechanisms for C5a-induced IL-8 production, which was 14-fold greater than that in unstimulated cells by 2 hours. IL-8 secretion was accompanied by accumulation of IL-8 mRNA in the cytosol and by nuclear expression of a κB DNA binding activity within 30 minutes. AP-1 but not NF-IL-6 DNA binding activity was also detected in C5a-stimulated PBMC; however, its delayed expression (maximal at 4 hours) suggested a less important role in the rapid production of IL-8. The correlation between C5a- induced κB binding activity and IL-8 gene expression was examined in the RAW264.7 macrophage cells using reporter genes directed by the κB sequence from IκBα and IL-8 promoter regions. C5a-induced reporter gene expression was abolished by introducing mutations into the κB sites and by coexpression of a dominant negative IκBα construct resistant to agonist-induced phosphorylation. Pertussis toxin, which ADP-ribosylates the G(i) proteins known to couple to the C5a receptor, produced minimal inhibition of C5a- induced IL-8 expression and had little effect on C5a-induced calcium mobilization in RAW264.7 cells. These results suggest that NF-κB activation is required for C5a-induced IL-8 gene expression and that this response is mediated primarily through a pertussis toxin-insensitive pathway.

Original languageEnglish (US)
Pages (from-to)3241-3249
Number of pages9
JournalBlood
Volume93
Issue number10
StatePublished - May 15 1999

Fingerprint

Interleukin-8
Gene expression
Chemical activation
Gene Expression
Pertussis Toxin
Reporter Genes
Blood Cells
Blood
Anaphylatoxin C5a Receptor
Phosphorylation
Macrophages
DNA
Chemotactic Factors
Transcription Factor AP-1
Genetic Promoter Regions
Cytosol
Adenosine Diphosphate
Interleukin-6
Genes
Calcium

ASJC Scopus subject areas

  • Biochemistry
  • Immunology
  • Hematology
  • Cell Biology

Cite this

NF-κB activation is required for C5a-induced interleukin-8 gene expression in mononuclear cells. / Hsu, Matthew H.; Wang, Meiying; Browning, Darren D; Mukaida, Naofumi; Ye, Richard D.

In: Blood, Vol. 93, No. 10, 15.05.1999, p. 3241-3249.

Research output: Contribution to journalArticle

Hsu, MH, Wang, M, Browning, DD, Mukaida, N & Ye, RD 1999, 'NF-κB activation is required for C5a-induced interleukin-8 gene expression in mononuclear cells', Blood, vol. 93, no. 10, pp. 3241-3249.
Hsu, Matthew H. ; Wang, Meiying ; Browning, Darren D ; Mukaida, Naofumi ; Ye, Richard D. / NF-κB activation is required for C5a-induced interleukin-8 gene expression in mononuclear cells. In: Blood. 1999 ; Vol. 93, No. 10. pp. 3241-3249.
@article{d4420908ad534edca801e5b54ef5929f,
title = "NF-κB activation is required for C5a-induced interleukin-8 gene expression in mononuclear cells",
abstract = "C5a, a potent peptide chemoattractant, stimulates interleukin-8 (IL-8) secretion from peripheral blood mononuclear cells (PBMC). Experiments were conducted to understand the mechanisms for C5a-induced IL-8 production, which was 14-fold greater than that in unstimulated cells by 2 hours. IL-8 secretion was accompanied by accumulation of IL-8 mRNA in the cytosol and by nuclear expression of a κB DNA binding activity within 30 minutes. AP-1 but not NF-IL-6 DNA binding activity was also detected in C5a-stimulated PBMC; however, its delayed expression (maximal at 4 hours) suggested a less important role in the rapid production of IL-8. The correlation between C5a- induced κB binding activity and IL-8 gene expression was examined in the RAW264.7 macrophage cells using reporter genes directed by the κB sequence from IκBα and IL-8 promoter regions. C5a-induced reporter gene expression was abolished by introducing mutations into the κB sites and by coexpression of a dominant negative IκBα construct resistant to agonist-induced phosphorylation. Pertussis toxin, which ADP-ribosylates the G(i) proteins known to couple to the C5a receptor, produced minimal inhibition of C5a- induced IL-8 expression and had little effect on C5a-induced calcium mobilization in RAW264.7 cells. These results suggest that NF-κB activation is required for C5a-induced IL-8 gene expression and that this response is mediated primarily through a pertussis toxin-insensitive pathway.",
author = "Hsu, {Matthew H.} and Meiying Wang and Browning, {Darren D} and Naofumi Mukaida and Ye, {Richard D.}",
year = "1999",
month = "5",
day = "15",
language = "English (US)",
volume = "93",
pages = "3241--3249",
journal = "Blood",
issn = "0006-4971",
publisher = "American Society of Hematology",
number = "10",

}

TY - JOUR

T1 - NF-κB activation is required for C5a-induced interleukin-8 gene expression in mononuclear cells

AU - Hsu, Matthew H.

AU - Wang, Meiying

AU - Browning, Darren D

AU - Mukaida, Naofumi

AU - Ye, Richard D.

PY - 1999/5/15

Y1 - 1999/5/15

N2 - C5a, a potent peptide chemoattractant, stimulates interleukin-8 (IL-8) secretion from peripheral blood mononuclear cells (PBMC). Experiments were conducted to understand the mechanisms for C5a-induced IL-8 production, which was 14-fold greater than that in unstimulated cells by 2 hours. IL-8 secretion was accompanied by accumulation of IL-8 mRNA in the cytosol and by nuclear expression of a κB DNA binding activity within 30 minutes. AP-1 but not NF-IL-6 DNA binding activity was also detected in C5a-stimulated PBMC; however, its delayed expression (maximal at 4 hours) suggested a less important role in the rapid production of IL-8. The correlation between C5a- induced κB binding activity and IL-8 gene expression was examined in the RAW264.7 macrophage cells using reporter genes directed by the κB sequence from IκBα and IL-8 promoter regions. C5a-induced reporter gene expression was abolished by introducing mutations into the κB sites and by coexpression of a dominant negative IκBα construct resistant to agonist-induced phosphorylation. Pertussis toxin, which ADP-ribosylates the G(i) proteins known to couple to the C5a receptor, produced minimal inhibition of C5a- induced IL-8 expression and had little effect on C5a-induced calcium mobilization in RAW264.7 cells. These results suggest that NF-κB activation is required for C5a-induced IL-8 gene expression and that this response is mediated primarily through a pertussis toxin-insensitive pathway.

AB - C5a, a potent peptide chemoattractant, stimulates interleukin-8 (IL-8) secretion from peripheral blood mononuclear cells (PBMC). Experiments were conducted to understand the mechanisms for C5a-induced IL-8 production, which was 14-fold greater than that in unstimulated cells by 2 hours. IL-8 secretion was accompanied by accumulation of IL-8 mRNA in the cytosol and by nuclear expression of a κB DNA binding activity within 30 minutes. AP-1 but not NF-IL-6 DNA binding activity was also detected in C5a-stimulated PBMC; however, its delayed expression (maximal at 4 hours) suggested a less important role in the rapid production of IL-8. The correlation between C5a- induced κB binding activity and IL-8 gene expression was examined in the RAW264.7 macrophage cells using reporter genes directed by the κB sequence from IκBα and IL-8 promoter regions. C5a-induced reporter gene expression was abolished by introducing mutations into the κB sites and by coexpression of a dominant negative IκBα construct resistant to agonist-induced phosphorylation. Pertussis toxin, which ADP-ribosylates the G(i) proteins known to couple to the C5a receptor, produced minimal inhibition of C5a- induced IL-8 expression and had little effect on C5a-induced calcium mobilization in RAW264.7 cells. These results suggest that NF-κB activation is required for C5a-induced IL-8 gene expression and that this response is mediated primarily through a pertussis toxin-insensitive pathway.

UR - http://www.scopus.com/inward/record.url?scp=0033561796&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033561796&partnerID=8YFLogxK

M3 - Article

C2 - 10233875

AN - SCOPUS:0033561796

VL - 93

SP - 3241

EP - 3249

JO - Blood

JF - Blood

SN - 0006-4971

IS - 10

ER -