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Int J Colorectal Dis. 2008 Mar;23(3):305-17. Epub 2007 Nov 28.

NF-kappaB-dependent synergistic regulation of CXCL10 gene expression by IL-1beta and IFN-gamma in human intestinal epithelial cell lines.

Author information

1
Centre for Internal Medicine, Department of Gastroenterology and Endocrinology, University of Göttingen, Goettingen, Germany.

Abstract

BACKGROUND AND AIMS:

Little is known about the intestinal epithelial expression and secretion of CXCL10 (IP-10), a chemokine involved in recruiting T cells and monocytes. We aimed to study CXCL10 gene expression and regulation by the pro-inflammatory cytokines interleukin (IL)-1beta, interferon (IFN)-gamma and tumour necrosis factor (TNF)-alpha in intestinal epithelial cell lines.

MATERIALS AND METHODS:

CXCL10 expression and secretion kinetics were assessed in Caco-2, HT-29 and DLD1 human colon epithelial cells, treated with IL-1beta, TNF-alpha, IFN-gamma alone or in combination with each other by real-time polymerase chain reaction (PCR), Northern blotting and enzyme-linked immunoabsorbent assay (ELISA). Transient transfections with TGL-IP10 (CXCL10 promoter) and TGL-IP10-kappaB2 mutant promoter and gelshifts and supershifts for nuclear factor (NF)-kappaB were also performed.

RESULTS:

Real-time PCRs and ELISA experiments revealed that IL-1beta was the strongest and earliest inducer of CXCL10 messenger ribonucleic acid (mRNA) expression and protein secretion in Caco-2 cell line, whereas INF-gamma had a delayed kinetics. There was a strong synergistic effect of either TNF-alpha or IL-1beta with IFN-gamma both on CXCL10 mRNA expression and protein secretion in all three cell lines. Real-time PCR and ELISA experiments using a specific NF-kappaB inhibitor and transfection experiments with a NF-kappaB-binding defective CXCL10 promoter construct revealed that the induction of CXCL10 by IL-1beta and its synergism with IFN-gamma is NF-kappaB dependent.

CONCLUSION:

These data demonstrate that in colonic epithelial cells, depending on the cellular context and utilizing the NF-kappaB pathway, IL-1beta alone and/or in synergism with IFN-gamma may play a major role in the induction of CXCL10.

PMID:
18046562
PMCID:
PMC2225996
DOI:
10.1007/s00384-007-0396-6
[Indexed for MEDLINE]
Free PMC Article

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