The effects of inhibitors of vacuolar acidification on the release of Listeria monocytogenes from phagosomes of Caco-2 cells

To evaluate the role of the acidic pH of phagosomes on the invasive ability and fate of Listeria monocytogenes within host cells, entry and replication of this gram-positive bacterium in a human enterocyte-like cell line (Caco-2) were investigated by a combination of biochemical and ultrastructural approaches, The effects of inhibitors of vacuolar acidification - the lipophilic weak base ammonium chloride, the carboxylic ionophore monensin and the vacuolar proton ATPase inhibitor bafilomycin A(1) - on the bacterial invasion pathway were analysed, These agents, which raise the intracellular vesicle acidic pH of living cells by different mechanisms, affected L. monocytogenes replication in Caco-2 cells, Bacteria internalised by bafilomycin-treated cells were unable to escape from phagosomes, as demonstrated by electronmicroscopy. The results provide evidence that low pH is required for efficient intracellular growth of L. monocytogenes.