Action of bacterial cytotoxins on normal mammalian cells and cells with altered membrane lipid composition.

Cytotoxic proteins produced by a number of bacteria, as well as one from a marine invertebrate, were tested for their ability to disrupt the permeability barrier of mammalian cells. Agents were tested individually and in combination shown to have synergistic disruptive actions on erythrocytes. Toxins included the lipid-hydrolyzing enzymes sphingomyelinases C and D and cholesterol oxidase, as well as the non-enzymatic agents, helianthus toxin, streptolysin O and saponin. Cells treated included cultured human skin fibroblasts, normal human erythrocytes and erythrocytes enhanced and depleted in membrane cholesterol. Fibroblasts were disrupted by helianthus toxin and by the combination of sphingomyelinase C and cholesterol oxidase. Thin layer chromatographic analysis of the treated cells confirmed the enzymatic alteration of membrane lipids by the lipid hydroxylases. Human erythrocytes having an increased content of membrane cholesterol were more sensitive than normal cells to agents which interact with membrane sterol. Conversely, cholesterol-depleted cells were more resistant to these as well as other agents. Results are discussed in relation to biochemical mechanisms of action of the agents tested, and to their possible significance in bacterial pathogenesis.