Direct measurement of phagosomal reactive oxygen by luminol-binding microspheres

A new method, utilizing microsphere-bound luminol, which makes possible the direct measurement of highly reactive oxygen within phagosomes, was studied. When Freund's complete adjuvant-elicited mouse peritoneal macrophages and luminol-binding microspheres were mixed, the microspheres were engulfed in macrophages and enclosed in phagosomes, where chemiluminescence (CL) was generated, showing the generation of highly reactive oxygen. The reactive oxygen could be quantitatively assayed by measuring the intensity of CL. The addition of cytochalasin B inhibited the CL. CL production by the thioglycollate-elicited macrophages was found to be only a ninth of that by Freund's complete adjuvant-elicited macrophages, though the phagocytic activities were almost equivalent in both cases.