Listeriolysin O affects barrier function and induces chloride secretion in HT-29/B6 colon epithelial cells.
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.
Institute of Clinical Physiology, Charité Berlin, Germany.
Jan F
JF
Richter
Alfred H
AH
Gitter
Dorothee
D
Günzel
Siegfried
S
Weiss
Walid
W
Mohamed
Trinad
T
Chakraborty
Michael
M
Fromm
Jörg D
JD
Schulzke
D017878:4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid / Q000494:pharmacology; D001427:Bacterial Toxins / Q000235:genetics / Q000494:pharmacology*; D001692:Biological Transport / Q000187:drug effects; D020013:Calcium Signaling / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002614:Chelating Agents / Q000494:pharmacology; D002712:Chlorides / Q000378:metabolism*; D005576:Colforsin / Q000494:pharmacology; D004533:Egtazic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D017097:Electric Impedance; D055724:Electrophysiological Phenomena / Q000187:drug effects; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism*; D019073:HT29 Cells; D006360:Heat-Shock Proteins / Q000235:genetics / Q000494:pharmacology*; D006460:Hemolysin Proteins / Q000235:genetics / Q000494:pharmacology*; D006801:Humans; D007413:Intestinal Mucosa / Q000378:metabolism*; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008353:Mannitol / Q000378:metabolism; D010539:Permeability / Q000187:drug effects; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D011994:Recombinant Proteins / Q000494:pharmacology; D012964:Sodium / Q000378:metabolism; D019284:Thapsigargin / Q000494:pharmacology; D013806:Theophylline / Q000494:pharmacology
D001427:Bacterial Toxins; D002614:Chelating Agents; D002712:Chlorides; D006360:Heat-Shock Proteins; D006460:Hemolysin Proteins; D047428:Protein Kinase Inhibitors; D011994:Recombinant Proteins; C070379:1,2-bis(2-aminophenoxy)ethane N,N,N',N'-tetraacetic acid acetoxymethyl ester; D005576:Colforsin; D008353:Mannitol; D004533:Egtazic Acid; D019284:Thapsigargin; D012964:Sodium; D013806:Theophylline; D007770:L-Lactate Dehydrogenase; D017878:4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid; C038705:hlyA protein, Listeria monocytogenes