A chemiluminescence procedure for determination of the release of myeloperoxidase from activated human neutrophils.
A chemiluminescence (CL) procedure was developed to determine the time course of the release of myeloperoxidase (MPO) from activated human neutrophils using two CL probes, luminol, and MCLA. Luminol-dependent CL (LDCL) and MCLA-dependent CL (MDCL) in a hypoxanthine (HX)/xanthine oxidase (XOD) system, both of which were completely inhibited by superoxide dismutase, were a linear function of the XOD concentration, with the relationship formula being LDCL = 0.003 x MDCL. Under the same conditions, MPO could enhance LDCL in a dose-dependent manner, without influencing MDCL. There was a linear correlation between the MPO concentrations and the values of (LDCL-0.003 x DCL) (coefficient of correlation = 0.004). This correlation made it possible to determine the release of MPO in the neutrophil/stimulus system by simultaneously monitoring the generation of LDCL and MDCL. By this CL procedure, it was revealed that there were variations in both neutrophil MPO releasing patterns and levels depending on the stimulating agent used.
Kitamura
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Department of Dermatology, Kurume University School of Medicine, Japan.
D000001:Calcimycin / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism; D006801:Humans; D008163:Luminescent Measurements; D018375:Neutrophil Activation; D009504:Neutrophils / Q000201:enzymology* / Q000502:physiology; D009195:Peroxidase / Q000378:metabolism*; D013481:Superoxides / Q000378:metabolism
D013481:Superoxides; D000001:Calcimycin; D009195:Peroxidase; D002118:Calcium