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Infection and Immunity, January 2005, p. 103-113, Vol. 73, No. 1
0019-9567/05/$08.00+0     doi:10.1128/IAI.73.1.103-113.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Role of EspA and Intimin in Expression of Proinflammatory Cytokines from Enterocytes and Lymphocytes by Rabbit Enteropathogenic Escherichia coli-Infected Rabbits

Karina Ramirez,1 Rocio Huerta,1 Eric Oswald,2 Carlos Garcia-Tovar,3 Jose Manuel Hernandez,1 and Fernando Navarro-Garcia1*

Department of Cell Biology, CINVESTAV-IPN, Mexico City,1 Morphology Unit, FES-Cuautitlán, UNAM, Cuatitlán Izcalli, Mexico,3 Unité Mixte de Microbiologie Moléculaire, Institut National de la Recherche Agronomique-Ecole Nationale Vétérinaire, Toulouse, France2

Received 3 August 2004/ Returned for modification 13 August 2004/ Accepted 7 September 2004

Enteropathogenic Escherichia coli (EPEC) produces attaching and effacing (A/E) lesions and watery diarrhea, both of which are intimin and EspA dependent. In this work, we explored the mucosal immune response by detecting cytokine induction in rabbits with diarrhea caused by rabbit EPEC (REPEC). Orally inoculated rabbits exhibited weight loss and mucosal inflammation, developed watery diarrhea, and died (day 7). At day 6 postinoculation, animals were analyzed for the induction of proinflammatory cytokines in enterocytes. The role of lymphocyte-dependent immunity was determined through the expression of proinflammatory cytokines by lymphocytes from Peyer's patches (PP) and the spleen. EspA and intimin mutants were used to explore the role of A/E lesions in the expression of these cytokines. REPEC-infected rabbit enterocytes showed increased interleukin 1ß (IL-1ß), IL-6, IL-8, and tumor necrosis factor alpha (TNF-{alpha}) mRNA expression, but that of anti-inflammatory IL-10 was increased only slightly. In contrast, intimin mutant-infected rabbits were unable to produce this proinflammatory cytokine profile but did produce a remarkable increase in IL-10 expression. Bacteria lacking EspA increased the expression of IL-8 and TNF-{alpha}, but that of IL-10 was increased only slightly. PP lymphocytes also produced proinflammatory cytokines, which were dependent on EspA (except for TNF-{alpha}) and intimin, while IL-10 was induced by EspA and intimin mutants. In contrast, spleen lymphocytes (systemic compartment) were unable to produce IL-1ß and TNF-{alpha}. These data show the importance of the proinflammatory cytokines secreted by enterocytes and those expressed locally by PP lymphocytes, which can activate effector mechanisms at the epithelium. Furthermore, this cytokine profile, including IL-6 and IL-1ß, which may be involved in the diarrhea produced by EPEC, depends on intimin.

* Corresponding author. Mailing address: Department of Cell Biology, CINVESTAV-IPN, Ap. Postal 14-740, 07000 México City, Mexico. Phone: (525) 5747-3990. Fax: (525) 5747-7081. E-mail: fnavarro{at}cell.cinvestav.mx.

Editor: J. T. Barbieri

Infection and Immunity, January 2005, p. 103-113, Vol. 73, No. 1
0019-9567/05/$08.00+0     doi:10.1128/IAI.73.1.103-113.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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