Transcriptional regulation of the IL-1β gene following GM-CSF and TNF stimulation of PMN

Marilyn Fernandez, John Walters, Phillip Marucha

Research output: Contribution to journalArticle

Abstract

Polymorphonuclear leukocytes (PMN) are recruited early at an inflammatory site and can modulate immune responses by secreting IL-1β. We had shown that TNF (10 ng/ml) and GM-CSF (10 ng/ml) induce IL-1β mRNA accumulation in PMN. Using nuclear run-on analysis, we had also shown that TNF and GM-CSF trigger IL-1β transcription independent of new protein synthesis. Here we show that activation of PKC and calcium mobilization are required for IL-1β transcription by TNF and GM-CSF. We have investigated the role of NF-kB, a transcription factor known to be involved in IL-1β transcription in monocytes, using electrophoretic mobility shift assays. Our results confirm that activation of NF-kB is triggered in mononuclear cells and undifferentiated HL-60 cells following cytokine stimulation. In PMN, there is a low level of constitutive NF-kB DNA binding activity, with minimal activation of NF-kB following stimulation with TNF, and no activation of NF-kB following stimulation with GM-CSF. This suggests that, in contrast to monocytes, activation of NF-kB does not play a major role in TNF or GM-CSF induction of IL-1β transcription in PMN.

Original languageEnglish (US)
JournalFASEB Journal
Volume10
Issue number6
StatePublished - 1996
Externally publishedYes

Fingerprint

transcription factor NF-kappa B
NF-kappa B
interleukin-1
Granulocyte-Macrophage Colony-Stimulating Factor
Interleukin-1
neutrophils
Neutrophils
Genes
Transcription
Chemical activation
transcription (genetics)
genes
monocytes
Monocytes
Electrophoretic mobility
HL-60 Cells
Electrophoretic Mobility Shift Assay
Assays
cytokines
Transcription Factors

ASJC Scopus subject areas

  • Agricultural and Biological Sciences (miscellaneous)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Cell Biology

Cite this

Transcriptional regulation of the IL-1β gene following GM-CSF and TNF stimulation of PMN. / Fernandez, Marilyn; Walters, John; Marucha, Phillip.

In: FASEB Journal, Vol. 10, No. 6, 1996.

Research output: Contribution to journalArticle

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AB - Polymorphonuclear leukocytes (PMN) are recruited early at an inflammatory site and can modulate immune responses by secreting IL-1β. We had shown that TNF (10 ng/ml) and GM-CSF (10 ng/ml) induce IL-1β mRNA accumulation in PMN. Using nuclear run-on analysis, we had also shown that TNF and GM-CSF trigger IL-1β transcription independent of new protein synthesis. Here we show that activation of PKC and calcium mobilization are required for IL-1β transcription by TNF and GM-CSF. We have investigated the role of NF-kB, a transcription factor known to be involved in IL-1β transcription in monocytes, using electrophoretic mobility shift assays. Our results confirm that activation of NF-kB is triggered in mononuclear cells and undifferentiated HL-60 cells following cytokine stimulation. In PMN, there is a low level of constitutive NF-kB DNA binding activity, with minimal activation of NF-kB following stimulation with TNF, and no activation of NF-kB following stimulation with GM-CSF. This suggests that, in contrast to monocytes, activation of NF-kB does not play a major role in TNF or GM-CSF induction of IL-1β transcription in PMN.

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