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Infection and Immunity, October 2009, p. 4406-4413, Vol. 77, No. 10
0019-9567/09/$08.00+0     doi:10.1128/IAI.00177-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

The Flagella of an Atypical Enteropathogenic Escherichia coli Strain Are Required for Efficient Interaction with and Stimulation of Interleukin-8 Production by Enterocytes In Vitro{triangledown}

Suely C. F. Sampaio,1 Tânia A. T. Gomes,1* Christophe Pichon,2,{dagger} Laurence du Merle,2,{dagger} Stéphanie Guadagnini,3 Cecilia M. Abe,4 Jorge L. M. Sampaio,5 and Chantal Le Bouguénec2,{dagger}

Departamento de Microbiologia, Imunologia e Parasitologia, Universidade Federal de São Paulo, Rua Botucatu 862, 3° andar, 04023-062 São Paulo, Brazil,1 Institut Pasteur, Unité de Pathogénie Bactérienne des Muqueuses, 28 rue du Dr Roux, F75724 Paris Cedex 15, France,2 Institut Pasteur, Plateforme de Microscopie Ultrastructurale, 25 rue du Dr Roux, F75724 Paris Cedex 15, France,3 Laboratório de Bacteriologia, Instituto Butantan, Avenida Vital Brazil, 1500, 05503-900 São Paulo, Brazil,4 Instituto Fleury de Ensino e Pesquisa, Avenida General Valdomiro de Lima, 508, 04344-903 São Paulo, Brazil5

Received 16 February 2009/ Returned for modification 29 March 2009/ Accepted 13 July 2009

The ability of some typical enteropathogenic Escherichia coli (EPEC) strains to adhere to, invade, and increase interleukin-8 (IL-8) production in intestinal epithelial cells in vitro has been demonstrated. However, few studies regarding these aspects have been performed with atypical EPEC (aEPEC) strains, which are emerging enteropathogens in Brazil. In this study, we evaluated a selected aEPEC strain (1711-4) of serotype O51:H40, the most prevalent aEPEC serotype in Brazil, in regard to its ability to adhere to and invade Caco-2 and T84 cells and to elicit IL-8 production in Caco-2 cells. The role of flagella in aEPEC 1711-4 adhesion, invasion, and IL-8 production was investigated by performing the same experiments with an isogenic aEPEC mutant unable to produce flagellin (FliC), the flagellum protein subunit. We demonstrated that this mutant (fliC mutant) had a marked decrease in the ability to adhere to T84 cells and invade both T84 and Caco-2 cells in gentamicin protection assays and by transmission electron microscopy. In addition, the aEPEC 1711-4 fliC mutant had a reduced ability to stimulate IL-8 production by Caco-2 cells in early (3-h) but not in late (24-h) infections. Our findings demonstrate that flagella of aEPEC 1711-4 are required for efficient adhesion, invasion, and early but not late IL-8 production in intestinal epithelial cells in vitro.

* Corresponding author. Mailing address: Departamento de Microbiologia, Imunologia e Parasitologia, Universidade Federal de São Paulo, Escola Paulista de Medicina, Rua Botucatu 862, 3° andar, 04023-062 São Paulo, Brazil. Phone: 55 11 5576 4537. Fax: 55 11 5572 4711. E-mail: tatg.amaral{at}unifesp.br

{triangledown} Published ahead of print on 20 July 2009.

Editor: J. B. Bliska

{dagger} Present address: Institut Pasteur, Biologie des Bactéries Pathogènes à Gram Positif, Batiment Duclaux, 75724, 28 rue du Dr Roux, Paris Cedex 15, France.

Infection and Immunity, October 2009, p. 4406-4413, Vol. 77, No. 10
0019-9567/09/$08.00+0     doi:10.1128/IAI.00177-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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