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Am J Physiol Lung Cell Mol Physiol (August 9, 2002). doi:10.1152/ajplung.00162.2002
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Articles in PresS, published online ahead of print August 9, 2002
Am J Physiol Lung Cell Mol Physiol, 10.1152/ajplung.00162.2002
Submitted on May 23, 2002
Accepted on August 3, 2002

Regulation of RANTES Promoter Activation in Alveolar Epithelial Cells Following Cytokine Stimulation

Antonella Casola1*, Allyne Henderson1, Tianshuang Liu1, Roberto P. Garofalo2, and Allan R. Brasier3

1 Department of Pediatrics, University of Texas Medical Branch, Galveston, TX, USA
2 Department of Pediatrics, University of Texas Medical Branch, Galveston, TX, USA; Department of Microbiology & Immunology, University of Texas Medical Branch, Galveston, TX, USA
3 Department of Internal Medicine, University of Texas Medical Branch, Galveston, TX, USA; Sealy Center for Molecular Sciences, University of Texas Medical Branch, Galveston, TX, USA

* To whom correspondence should be addressed. E-mail: ancasola{at}utmb.edu.

RANTES is a member of the CC chemokine family of proteins, implicated in a variety of diseases characterized by lung eosinophilia and inflammation, strongly produced by stimulated airway epithelial cells. Since cytokines like TNF-{alpha} and IFN-{gamma}have been shown to enhance RANTES induction in airway epithelial cells, and RANTES gene expression appears to be differentially regulated depending on the cell type and the stimulus applied, in this study we have elucidated the mechanisms that operate to control RANTES induction upon exposure to TNF-{alpha} and IFN-{gamma} alone or in combination. Our results indicate that TNF-{alpha} and IFN-{gamma} synergistically induce RANTES protein secretion and mRNA expression. However RANTES transcription is activated only following stimulation with TNF-{alpha}, but not with IFN-{gamma}. Promoter deletion and mutagenesis experiments indicate that the NF-{kappa}B site is the most important cis-regulatory element controlling TNF-induced RANTES transcription, although the NF-IL6 binding site, the cAMP Responsive Element (CRE) and the Interferon Stimulated Responsive Element (ISRE) also play a significant role. TNF-{alpha} stimulation induces nuclear translocation of Interferon Regulatory Factor (IRF)-3, which in viral infection binds the RANTES ISRE and is necessary for activation of RANTES transcription. However, TNF-induced IRF-3 translocation does not result in IRF-3 binding to the RANTES ISRE. Furthermore, while viral infection can activate an ISRE-driven promoter, TNF cannot, indicating that RANTES gene enhancers are controlled in a stimulus-specific fashion. Identification of the molecular mechanisms involved in RANTES gene expression is fundamental for developing strategies to modulate lung inflammatory responses.




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