Pathogenetic Impact of Vacuolar Degeneration by Accelerated Transport of Helicobacter pylori VacA

Vacuolar degeneration of the gastric epithelial cells is a characteristic feature of the derangement of mucosa where Helicobacter pylori colonizes, and H. pylori vacuolating cytotoxin (VacA) has been suggested to playa key role in it. To elucidate the VacA-involved degenerative mechanism, VacA was purified, and its impact on degeneration of HeLa cells was determined. In the presence of ammonium chloride, cell vacuolation by VacA was dose-and time-dependent, however, no detectable degeneration of the cells was observed with the VacA concentration tested. A further increase of vacuolation was shown in cells pre-treated with diethyl pyrocarbonate (DEPC) , and this resulted in a change of the cell morphology to become spherical. Similar phenomena were also observed when HeLa cells were co-cultivated with intact H. pylori cells. It was remarkable to note that the degree of growth inhibition was proportional to the increase in vacuole formation, suggesting that the vacuolation rate would be critical for cell degeneration. Surprisingly, although VacA was itself inhibited by DEPC, its uptake was markedly increased by this agent, similar to that found in cells with Nabutyrate. These data indicate that the cell's tolerance of VacA transport may be critical for vacuolar degeneration and may be changeable during H. pylori inhabitation.