Particle-induced myeloperoxidase release in serially diluted whole blood quantifies the number and the phagocytic activity of blood neutrophils and opsonization capacity of plasma.
Luminol-amplified chemiluminescence (CL) from phagocytes has previously been shown to be almost completely dependent on the release of myeloperoxidase (MPO) from azurophilic granules. We measured the luminol-amplified chemiluminescence response (WBCL) by using serially diluted whole blood. In these experiments, non-opsonized and serum-opsonized zymosan (NWBCL and OWBCL, respectively) were used concurrently as phagocytosable particles. We found two whole-blood dilution ranges with clinical significance: first, <0.04% of whole blood in the reaction volume, where MPO released by the zymosan-activated leukocyte population came almost totally from neutrophils and the OWBCL response could be exploited as a measure of a neutrophil count in a given blood specimen, despite the pathophysiological state of the host. In contrast, the NWBCL response was two-fold in blood samples from bacterial infection patients compared to those of controls and patients with viral infection, suggesting the use of NWBCL for the differential diagnosis of bacterial infections from viral infections; second, 0.16-1.2% of whole blood in the reaction volume, where the opsonization capacity of plasma (OC(50)) can be determined. We also found that at whole blood content >0.04%, erythrocytes quickly start to absorb chemiluminescence light, and that at whole blood content >1.2%, plasma proteins, most probably albumin and fibrinogen, start to inhibit MPO release.
Lilius
Esa-Matti Eino
EM
University of Turku, Department of Biochemistry, Vatselankatu 2, 20014 Turku, Finland. esalil@utu.fi
Nuutila
Jari Timo Juhani
JT
D001424:Bacterial Infections / Q000097:blood; D004769:Enterovirus Infections / Q000097:blood; D006801:Humans; D008163:Luminescent Measurements; D008165:Luminol / Q000494:pharmacology; D009000:Monocytes / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D009504:Neutrophils / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D009895:Opsonin Proteins / Q000097:blood; D009195:Peroxidase / Q000097:blood*; D010586:Phagocytes / Q000378:metabolism / Q000502:physiology*; D013997:Time Factors; D015054:Zymosan / Q000494:pharmacology
D009895:Opsonin Proteins; D008165:Luminol; D015054:Zymosan; D009195:Peroxidase