Infection of rabbit kidney cells (RK13) by enteropathogenic Escherichia coli as a model to study the dynamics of actin cytoskeleton

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Abstract:

Enteropathogenic Escherichia coli (EPEC) colonizes the intestinal mucosa and causes a cell lesion known as attachment and effacement (A/E) lesion. The molecular mechanisms for A/E lesions include injection of Tir, which is a receptor for an adhesin named intimin. The Tir-intimin interaction causes rearrangement of the cytoskeleton forming actin-rich structures called pedestals. Unfortunately, the formation of the A/E lesions and the dynamics of the actin cytoskeleton during this rearrangement induced by EPEC cannot be studied in the natural host. However, there are EPEC strains that infect rabbit (REPEC) that are genetically and pathologically similar to EPEC. Here, we used REPEC for the infection of rabbit kidney epithelial cells, line RK13, as a model to understand the actin cytoskeleton dynamics during pedestal formation. Actin-rich pedestal formation during the infection of RK13 cells by REPEC was analyzed by electron and confocal microscopy. The kinetics of infection along with the use of antibiotics for eliminating the bacteria, as well as reinfection, evidenced the plasticity of the actin cytoskeleton during pedestal formation. Thus, this model is a helpful tool for studying the dynamics of actin cytoskeleton and for correlating the data with those observed in in vivo models in rabbits experimentally infected with REPEC.

Escherichia coli entérophathogène (EPEC) colonise la muqueuse intestinale et cause des lésions cellulaires appelées lésions d’attachement et d’effacement (A/E). Les mécanismes moléculaires associés aux lésions incluent l’implication de Tir, le récepteur d’une adhésine, l’intimine. L’interaction Tir-intimine cause un réarrangement du cytosquelette formant une structure riche en actine, le piédestal. Malheureusement, la formation de lésions A/E et la dynamique du cytosquelette d’actine lors du réarrangement induit par les EPEC ne peuvent être étudiées chez l’hôte naturel. Cependant, il existe des souches d’EPEC qui peuvent infecter le lapin (REPEC) et qui sont génétiquement et pathologiquement similaires aux EPEC. Nous avons utilisé ici des REPEC pour infecter des cellules épithéliales rénales de lapin, les cellules de la lignée RK13, un modèle permettant de comprendre la dynamique du cytosquelette d’actine lors de la formation de piédestaux. La formation d’un piédestal riche en actine lors de l’infection des cellules RK13 par les REPEC a été analysée par microscopie électronique et par microscopie confocale. La cinétique d’infection et l’utilisation d’antibiotiques pour éliminer les bactéries, ainsi que la réinfection ont mis en évidence la plasticité du cytosquelette d’actine lors de la formation du piédestal. Ainsi, ce modèle constitue un outil pratique pour étudier la dynamique du cytosquelette d’actine et pour corréler les résultats avec les observations faites dans des modèles in vivo chez le lapin infecté par les REPEC.

Document Type: Research Article

Publication date: September 1, 2008

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  • Published since 1954, this monthly journal contains new research in the field of microbiology including applied microbiology and biotechnology; microbial structure and function; fungi and other eucaryotic protists; infection and immunity; microbial ecology; physiology, metabolism and enzymology; and virology, genetics, and molecular biology. It also publishes review articles and notes on an occasional basis, contributed by recognized scientists worldwide.
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