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Infection and Immunity, March 2003, p. 1068-1074, Vol. 71, No. 3
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.3.1068-1074.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

The Afa/Dr Adhesins of Diffusely Adhering Escherichia coli Stimulate Interleukin-8 Secretion, Activate Mitogen-Activated Protein Kinases, and Promote Polymorphonuclear Transepithelial Migration in T84 Polarized Epithelial Cells

Fréderic Bétis,1 Patrick Brest,1 Véronique Hofman,2 Julie Guignot,3 Marie-Françoise Bernet-Camard,3 Bernard Rossi,1 Alain Servin,3 and Paul Hofman1,2*

INSERM 364,1 Laboratoire d'Anatomie Pathologique, Faculté de Médecine, 06107 Nice Cédex 02,2 INSERM 510, Faculté de Pharmacie, Paris XI, F-92296 ChÂtenay-Malabry, France3

Received 7 August 2002/ Returned for modification 31 October 2002/ Accepted 10 December 2002

Afa/Dr diffusely adhering Escherichia coli (Afa/Dr DAEC) strains cause symptomatic urinary tract and intestinal infections. The proinflammatory effects of Afa/Dr DAEC strains in vitro have been not investigated to date. In the present study, we used confluent polarized monolayers of intestinal cell line T84 to evaluate the consequences of epithelial infection by Afa/Dr DAEC strains in terms of proinflammatory response. Polymorphonuclear leukocyte (PMNL) migration across the epithelial barrier was induced after incubation of the T84 monolayers with the wild-type Afa/Dr DAEC strain C1845 harboring the fimbrial F1845 adhesin and strain IH11128 harboring the Dr hemagglutinin, and the E. coli laboratory strain HB101 was transformed with the pSSS1 plasmid, producing Afa/Dr F1845 adhesin. PMNL migrations were correlated with a basolateral secretion of interleukin-8 by T84 cells and were abolished after incubation of epithelial cells with an anti-decay accelerating factor (DAF) antibody that recognized the short consensus repeat 3 domain of DAF (monoclonal antibody 1H4). Moreover, Afa/Dr DAEC strains induced tyrosine phosphorylation of several T84 proteins and activated the mitogen-activated protein kinases (ERK1/2 mitogen-activated protein, P38, and Jun-C kinases). These data demonstrated for the first time that, in vitro, Afa/Dr DAEC strains exert a proinflammatory signal in intestinal epithelial cells.

* Corresponding author. Mailing address: INSERM unité 364, IFR 50, Faculté de Médecine, Avenue de Valombrose, 06107 Nice, Cédex 02, France. Phone: 33-4-93-37-77-59. Fax: 33-4-93-81-94-56. E-mail: hofman{at}unice.fr.

Editor: A. D. O'Brien

Infection and Immunity, March 2003, p. 1068-1074, Vol. 71, No. 3
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.3.1068-1074.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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