BOCCO JOSE LUIS
Congresos y reuniones científicas
Título:
El Tor haemolysin from Vibrio cholerae induces apoptosis and increases intracellular Ca2+ levels in human intestinal cells.
Autor/es:
SAKA, HÉCTOR A.; BIDINOST, CARLA; BONACCI, GUSTAVO; CHIABRANDO, GUSTAVO; BOCCO, JOSÉ LUIS
Lugar:
Iguazu, Misiones - ARGENTINA
Reunión:
Congreso; XL Reunión Anual de la Sociedad Argentina de Investigaciones en Bioquímica y Biología Molecular.; 2004
Institución organizadora:
Sociedad Argentina de Investigaciones en Bioquímica y Biología Molecular.
Resumen:

V.cholerae O1 biotype El Tor and most of V.cholerae non-O1 / non-O139 (VCN) produce El Tor Haemolysin (ETH), a pore forming toxin encoded by the hlyA gene. In previous studies we have shown that ETH could be involved in the diarrhea caused by a VCN clinical isolate lacking cholera toxin (VCN CT-). Additionally, we demonstrated ETH-induced apoptosis on Cos-7 cells.To explore the potential involvement of apoptosis in the pathogenesis caused by ETH on human intestinal cells,we exposed undifferentiated Caco-2 and differentiated C2BBe1 to sterile culture supernatants from VCN CT-or from its isogenic hlyA null mutant. At different times post-incubation, caspase-3 activation, internucleosomal DNA fragmentation and increased sub-G1 fraction of fragmented nuclei were detected by a Caspase-3 apoptosis detection kit, agarose gel electrophoresis/ethidium bromide staining and flow cytometry/propidium iodide staining, respectively. To investigate the role of second messengers involved in response to ETH, changes in Caco-2 intracellular Ca 2+  levels were measured after exposition to ETH by flow cytometry using Fluo3-AM.After 2 and 4 h of exposition to 200 ng/mL of ETH, intracellular Ca 2+ increased 22,0% and 36,6%,respectively. These results demonstrate that when added to human intestinal cells, ETH is able of causing cell death via apoptosis and that an intracellular increase in Ca 2+  concentration occurs in response to this toxin.