leukotoxin and Leukemia--Promyelocytic--Acute

The effects of Ca2+ on the binding of Actinobacillus actinomycetemcomitans leukotoxin (LT) to LT-susceptible promyelocytic leukemia HL-60 cells and the consequent cytotoxicity were investigated in comparison with those on LT-resistant erythroleukemia K-562 cells. The total amount of LT bound to the HL-60 and K-562 cells, as determined by ELISA using anti-LT rabbit serum, was essentially the same for both cells. The percentage of bound LT decreased in the presence of phosphatidylcholine (PC) was also almost equal for both cells. In contrast, EGTA added in the presence of PC enhanced the decrease of bound LT significantly from HL-60, but not from K-562. By monitoring the fluorescence of Fura-2 and of fluorescein as a measure for the change in intracellular calcium ions ([Ca2+]i) and cytotoxic activity, respectively, we showed that the exposure of HL-60 to LT induced the [Ca2+]i increase followed by the cell death, whereas this was distinctly not the case for K-562. These results indicate that a Ca(2+)-dependent LT-binding mechanism is possibly the step characteristic to LT-susceptible HL-60 cells.

Topics: Aggregatibacter actinomycetemcomitans; Binding Sites; Calcium; Cell Death; Cytotoxins; Egtazic Acid; Enzyme-Linked Immunosorbent Assay; Exotoxins; Humans; Leukemia, Erythroblastic, Acute; Leukemia, Promyelocytic, Acute; Phosphatidylcholines; Tumor Cells, Cultured