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Infection and Immunity, February 2005, p. 712-721, Vol. 73, No. 2
0019-9567/05/$08.00+0     doi:10.1128/IAI.73.2.712-721.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

HtrA Stress Protein Is Involved in Intramacrophagic Replication of Adherent and Invasive Escherichia coli Strain LF82 Isolated from a Patient with Crohn's Disease

Marie-Agnès Bringer,1 Nicolas Barnich,1 Anne-Lise Glasser,1 Olivier Bardot,2 and Arlette Darfeuille-Michaud1*

Pathogénie Bactérienne Intestinale, Laboratoire de Bactériologie, Université d'Auvergne, CBRV,1 INSERM U384, Clermont-Ferrand, France2

Received 14 May 2004/ Returned for modification 16 August 2004/ Accepted 1 October 2004

Adherent and invasive Escherichia coli (AIEC) bacteria isolated from Crohn's disease patients are able to greatly replicate within macrophages without escaping from the phagosome and without inducing macrophage death. In the present study, evidence is provided that in AIEC strain LF82 the htrA gene encoding the stress protein HtrA is essential for intracellular replication within J774-A1 macrophages. Deletion of the htrA gene in strain LF82 induced increased sensitivity of the isogenic mutant to oxidative stress caused by hydrogen peroxide and a reduced rate of growth in an acid and nutrient-poor medium partly reproducing the microenvironment of the phagosome. In vitro experiments using an LF82 htrA gene promoter fusion with the lacZ gene revealed a 38-fold activation of the promoter in AIEC LF82 intramacrophagic bacteria. The CpxRA two-component signaling pathway was not involved in this activation. In addition, the activation of the LF82 htrA gene promoter was not observed in the nonpathogenic E. coli K-12 intramacrophagic bacteria, indicating that the AIEC LF82 genetic background is crucial for induction of htrA gene transcription during phagocytosis.

* Corresponding author. Mailing address: Pathogénie Bactérienne Intestinale, Laboratoire de Bactériologie, Université d'Auvergne, CBRV, 28 Place Henri Dunant, 63001 Clermont-Ferrand, France. Phone: 33 4 73 17 79 97. Fax: 33 4 73 17 83 71. E-mail: arlette.darfeuille-michaud{at}u-clermont1.fr.

Editor: V. J. DiRita

Infection and Immunity, February 2005, p. 712-721, Vol. 73, No. 2
0019-9567/05/$08.00+0     doi:10.1128/IAI.73.2.712-721.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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