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Infection and Immunity, May 2002, p. 2304-2310, Vol. 70, No. 5
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.5.2304-2310.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Enterohemorrhagic Escherichia coli Infection Induces Interleukin-8 Production via Activation of Mitogen-Activated Protein Kinases and the Transcription Factors NF-{kappa}B and AP-1 in T84 Cells

Stephanie Dahan,1 Valere Busuttil,2 Veronique Imbert,2 Jean-Francois Peyron,2 Patrick Rampal,1 and Dorota Czerucka1*

Laboratoire de Gastroenterologie et Nutrition,1 INSERM U526 Activation des cellules hematopoïetiques, IFR50, Faculte de Medecine, Universite de Nice-Sophia Antipolis, 06107 Nice cedex 2, France2

Received 16 November 2001/ Accepted 8 January 2002

Enterohemorrhagic Escherichia coli (EHEC) infections are associated with hemorrhagic colitis and the hemolytic-uremic syndrome (HUS). In vivo, elevated plasma levels of the proinflammatory cytokine interleukin-8 (IL-8) in EHEC-infected children are correlated with a high risk of developing HUS. As IL-8 gene transcription is regulated by the transcription factors NF-{kappa}B and AP-1, we analyzed the role of these factors in the regulation of IL-8 production after infection of the epithelial intestinal T84 cell line by EHEC. By 6 h of infection, EHEC had induced significant secretion of IL-8 (35.84 ± 6.76 ng/ml versus 0.44 ± 0.04 ng/ml in control cells). EHEC induced AP-1 and NF-{kappa}B activation by 3 h of infection. Moreover, the three mitogen-activated protein kinases (MAPK) (ERK1/2, p38, and JNK) were phosphorylated in EHEC-infected T84 cells concomitant with induction of AP-1 DNA binding activity, and I{kappa}B-{alpha} was phosphorylated and then degraded concomitant with induction of NF-{kappa}B DNA binding activity. Pretreatment of cells with the highly specific MEK1/2 inhibitor U0126, the p38 inhibitor SB203580, and/or the proteasome inhibitor ALLN led to inhibition of the IL-8 secretion induced in EHEC-infected T84 cells. These findings demonstrate that (i) EHEC can induce in vitro a potent proinflammatory response by secretion of IL-8 and (ii) the secretion of IL-8 is due to the involvement of MAPK, AP-1, and NF-{kappa}B signaling pathways.

* Corresponding author. Mailing address: Laboratoire de Gastroenterologie et Nutrition, Universite de Nice-Sophia Antipolis, 28 avenue de Valombrose, 06107 Nice cedex 2, France. Phone: (33) 4 93 37 76 95. Fax: (33) 4 93 81 77 10. E-mail: czerucka{at}unice.fr.

Editor: A. D. O'Brien

Infection and Immunity, May 2002, p. 2304-2310, Vol. 70, No. 5
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.5.2304-2310.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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