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Am J Physiol Gastrointest Liver Physiol 296: G1350-G1359, 2009. First published April 16, 2009; doi:10.1152/ajpgi.00040.2009
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MUCOSAL BIOLOGY

Listeriolysin O affects barrier function and induces chloride secretion in HT-29/B6 colon epithelial cells

Jan F. Richter,1 Alfred H. Gitter,1,4 Dorothee Günzel,1 Siegfried Weiss,5 Walid Mohamed,3 Trinad Chakraborty,3 Michael Fromm,1 and Jörg D. Schulzke2

1Institute of Clinical Physiology and 2Department of Gastroenterology, Charité Berlin, Germany; 3Institute of Medical Microbiology, Justus Liebig University, Giessen, Germany, and 5Helmholtz Centre for Infection Research, Braunschweig, Germany; and 4Division of Medical Engineering/Biotechnology, University of Applied Sciences, Jena, Germany

Submitted 22 January 2009 ; accepted in final form 7 April 2009

Listeria monocytogenes is a food-borne pathogen, which is able to induce diarrhea when residing in the intestine. We studied the effect of listeriolysin O (LLO), an extracellular virulence factor of L. monocytogenes, on intestinal transport and barrier function in monolayers of HT-29/B6 human colon cells using the Ussing technique to understand the pathomechanisms involved. Mucosal addition of LLO, but not a LLO mutant, induced a dose- and pH-dependent increase in short-circuit current (ISC). Sodium and chloride tracer flux and DIDS sensitivity studies revealed that ISC was mainly due to electrogenic chloride secretion. Barrier function was impaired by LLO, as assessed by transepithelial resistance (Rt) and mannitol flux measurements. Intracellular signal transduction occurred through Ca2+ release from intracellular stores and PKC activation. In conclusion, listeriolysin induces chloride secretion and perturbs epithelial barrier function, thus potentially contributing to Listeria-induced diarrhea.

listeria; diarrhea; transepithelial resistance; HT-29/B6 cells; calcium


Address for reprint requests and other correspondence: Jörg D. Schulzke, Dept. of Gastroenterology, Hindenburgdamm 30, Berlin, 12200, Germany (e-mail: joerg.schulzke{at}charite.de)






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