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Phorbol dibutyrate induced a nitroblue tetrazolium-reducing reaction in differentiated HL-60 cells, which was inhibited by protein kinase inhibitors such as staurosporine and H-7. ID50 of staurosporine and H-7 were 1.4 ng/ml and 0.19 mM, respectively. When tautomycin, an inhibitor of protein phosphatases, was added with the kinase inhibitors, the nitroblue tetrazolium-reducing reaction again appeared. ID50 of staurosporine was 510 ng/ml in the presence of tautomycin. Tautomycin itself weakly induced the reaction, which was inhibited by kinase inhibitors. Such a competitive effect between tautomycin and staurosporine was not observed in a cell-free system of protein kinase C. Okadaic acid had the same effect as tautomycin. The similar results were obtained when respiratory burst was quantitated by measuring H2O2 produced by canine peripheral neutrophils. The mechanism of competitive effect of tautomycin and staurosporine on respiratory burst is discussed.

Respiratory burst induced by phorbol ester in the presence of tautomycin, a novel inhibitor of protein phosphatases.