Enterotoxic effect of the vacuolating toxin produced by Helicobacter pylori in Caco-2 cells.
Preliminary clinical evidence suggests that Helicobacter pylori may be associated with diarrhea through its vacuolating toxin (VacA). To establish whether VacA induces intestinal secretion, epithelial damage, or both, purified pH-activated VacA was added to Caco-2 cell monolayers mounted in Ussing chambers, and electrical parameters were monitored. Mucosal addition of VacA induced an increase in short circuit current, consistent with enterotoxic effect. The effect was time- and dose-dependent and saturable. It was not found if the toxin was not pH-activated, added to the serosal side, or preheated. In cells preloaded with the Ca2+ buffering compound BAPTA/AM or with the Cl- channel inhibitor 5-nitro-2-3-(3-phenylpropylamino)benzoic acid, short circuit current did not change, indicating that VacA induces activation of Ca2+-dependent Cl- channels. VacA did not show cytopathic effects, as judged by tissue resistance. These results support the hypothesis that H. pylori may be associated with diarrhea through production of VacA.
Department of Pediatrics, University Federico II, Naples, Italy. alfguari@cds.unina.it
A
A
Guarino
M
M
Bisceglia
R B
RB
Canani
M C
MC
Boccia
G
G
Mallardo
E
E
Bruzzese
P
P
Massari
R
R
Rappuoli
J
J
Telford
D001426:Bacterial Proteins / Q000008:administration & dosage / Q000096:biosynthesis / Q000633:toxicity*; D001427:Bacterial Toxins / Q000008:administration & dosage / Q000096:biosynthesis / Q000633:toxicity*; D001692:Biological Transport / Q000187:drug effects; D018938:Caco-2 Cells / Q000187:drug effects; D003603:Cytotoxins / Q000008:administration & dosage / Q000096:biosynthesis / Q000633:toxicity*; D004305:Dose-Response Relationship, Drug; D004533:Egtazic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D017097:Electric Impedance; D016480:Helicobacter pylori / Q000378:metabolism*; D006801:Humans; D007202:Indicators and Reagents / Q000494:pharmacology; D007422:Intestines / Q000187:drug effects*; D014617:Vacuoles / Q000187:drug effects
D001426:Bacterial Proteins; D001427:Bacterial Toxins; D003603:Cytotoxins; D007202:Indicators and Reagents; C074689:VacA protein, Helicobacter pylori; D004533:Egtazic Acid; C025603:1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid