HEMOLYSIS OF RABBIT ERYTHROCYTES BY PURIFIED STAPHYLOCOCCAL ALPHA-TOXIN II

Cooper, Louis Z. (New England Center Hospital, Boston, Mass.), Morton A. Madoff, and Louis Weinstein. Hemolysis of rabbit erythrocytes by purified staphylococcal alpha-toxin. II. Effect of inhibitors on the hemolytic sequence. J. Bacteriol. 87:136–144. 1964.—Study of the time course of hemolysis of rabbit erythrocytes by purified staphylococcal alpha-lysin revealed that the specific toxin-red cell reaction occurs during the prelytic period. This reaction could be prevented or decreased by alpha-lysin antitoxin added early, but not by antitoxin added at the end of the prelytic phase or at any time thereafter. In contrast, hemolysis is suppressed temporarily by sucrose and permanently by polyethylene glycol, even when these are added during the period of rapid release of hemoglobin. When sucrose is present together with alpha-lysin and red cells only during the prelytic period, and when the cells are then washed and resuspended in phosphate-buffered saline, their subsequent hemolysis is not altered by the presence of the sugar. This is not so when antitoxin is employed. When erythrocytes are laked by a measured excess of alpha-lysin, only a portion of the original hemolytic activity can be recovered. Repeated exposure of lysin to red cells produces a loss of activity represented by a linear function when logs of residual activity are plotted sequentially. Once alpha-lysin has reacted with red cells, it does not appear to be available for attachment to other erythrocytes.