Preliminary incubation of peritoneal macrophages with binase in a concentration of 100 mkg/ml (and more) results in the reduction of the macrophages' death by necrosis induced by hydrogen peroxide. In this case the cells mostly enter apoptosis. The protection effect of binase may be a consequence of a stabilizing effect of binase on cytoplasmic membrane. The possibility of the lipid bilayer stabilization is indicated by the change in DPPC phase transition parameters in the presence of binase, which have been determined in a model experiment by differential scanning calorimetry. However, the influence of binase on the transmission of H2O2-induced pro-apoptotic signals should not be ruled out.

Effects of binase (RNAse of Bacillus intermedius, 40 and 400 μg/ml) on reactive oxygen species (ROS) generation by human peripheral blood granulocytes and monocytes were tested by fl ow cytometry. We found that binase selectively affected E. coli- and phorbol myristate acetate (PMA) – induced oxidative burst in granulocytes and did not infl uence ROS generation in monocytes. The response pattern of granulocytes depends on the ROS inducer and the binase concentration. In a setting of E coli-induced oxidative burst, binase upregulated both the ROS-producing granulocyte subset and the relative ROS generation per a single cell in a concentration-dependent manner. In the PMA-stimulated granulocytes, there was a tendency for differential concentration-dependent effects of binase on intracellular ROS generation.