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

• Published in: American journal of physiology: Gastrointestinal and liver physiology 2009-06, Vol.296 (6), p.G1350-G1359
• Main Authors: Richter, Jan F, Gitter, Alfred H, Günzel, Dorothee, Weiss, Siegfried, Mohamed, Walid, Chakraborty, Trinad, Fromm, Michael, Schulzke, Jörg D
• Format: Article
• Language: English
• Subjects: 4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid - pharmacology; Bacteria; Bacterial Toxins - genetics; Bacterial Toxins - pharmacology; Biological Transport - drug effects; Calcium Signaling - drug effects; Cell Survival - drug effects; Cells; Chelating Agents - pharmacology; Chlorides - metabolism; Colforsin - pharmacology; Colon; Diarrhea; Digestive system; Egtazic Acid - analogs & derivatives; Egtazic Acid - pharmacology; Electric Impedance; Electrophysiological Phenomena - drug effects; Epithelial Cells - drug effects; Epithelial Cells - metabolism; Epithelial Cells - secretion; Heat-Shock Proteins - genetics; Heat-Shock Proteins - pharmacology; Hemolysin Proteins - genetics; Hemolysin Proteins - pharmacology; HT29 Cells; Humans; Intestinal Mucosa - metabolism; L-Lactate Dehydrogenase - metabolism; Mannitol - metabolism; Permeability - drug effects; Protein Kinase Inhibitors - pharmacology; Recombinant Proteins - pharmacology; Signal transduction; Sodium - metabolism; Thapsigargin - pharmacology; Theophylline - pharmacology
• ISSN: 0193-1857; 1522-1547
• DOI: 10.1152/ajpgi.00040.2009

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 (I(SC)). Sodium and chloride tracer flux and DIDS sensitivity studies revealed that I(SC) was mainly due to electrogenic chloride secretion. Barrier function was impaired by LLO, as assessed by transepithelial resistance (R(t)) and mannitol flux measurements. Intracellular signal transduction occurred through Ca(2+) 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.