Comparative studies on the oxidative processes during phagocytosis measured by luminol-dependent chemiluminescence

Four different methods, using luminol-enhanced chemiluminescence, are described for assessment of the oxidative processes associated with the phagocytosis of opsonized zymosan by granulocytes. The techniques described in the literature using whole blood as a substrate gave unreliable results due to quench effects of contaminating red blood cells, unpredictable serum interferences, or varying ratios of granulocytes and zymosan. In order to avoid these side effects, rapid and simple methods for the isolation of the granulocytes were applied, e.g. lysis of erythrocytes by ammonium chloride, or the isolation of granulocytes on a percoll gradient. The latter procedure proved to be reliable, simple and sufficiently quick for a routine application.