Summary
The oxidative metabolism, and thus the bactericidal capacity, of neutrophil granulocytes can be evaluated by measurement of the luminol chemiluminescence during phagocytosis. The method described requires only a small number of separated cells (0.3×106); continuous measurement in the physiologic temperature range is possible.
The procedure is described and results of polymorphonuclear leukocytes from healthy adults and umbilical cord blood from healthy newborns are given. Quantitative variations in the single components (number of granulocytes, number of phagocytic paticles, concentration of luminol) and the measuring volume are investigated. The significance of the chosen parameters “photon emission during defined time of measurement” and “time of maximum photon emission per unit time” used for evaluation is shown by comparison of the results from the influence of phorbol myristate acetate on the luminol chemiluminescence of phagocytosing granulocytes. The method described enables the evaluation of the capacity and the temporal course of stimulation of the oxidative metabolism of polymorphonuclear granulocytes in phagocytosis.
Zusammenfassung
Die Messung der Luminol-Chemilumineszenz nach Stimulation durch Phagocytose erlaubt die Beurteilung des oxidativen Metabolismus und damit der O2-abhängigen Bakterizidie von neutrophilen Granulocyten. Die beschriebene Methode ermöglicht Messungen der Luminol-Chemilumineszenz in einer geringen Zahl separierter neutrophiler Granulocyten (0,3×106) und kontinuierliche Messungen im physiologischen Temperaturbereich.
Das methodische Vorgehen wird beschrieben und Ergebnisse der Granulocyten gesunder Erwachsener wie aus Nabelschnurblut gesunder Neugeborener angegeben. Untersuchungen mit quantitativen Variationen der Einzelkomponenten (Zahl der Granulocyten, Zahl der Phagocytatpartikel, Luminol-Konzentration) und des Meßvolumens wurden durchgeführt. Die Bedeutung der zur Beurteilung herangezogenen Parameter „Summe der während definierter Meßdauer emittierten Photonen“ und „Zeitpunkt der maximalen Photonenemission pro Zeiteinheit“ wird an den Beispielen der Ergebnisse von Granulocyten gesunder Erwachsener und aus Nabelschnurblut wie an der Wirkung von Phorbol Myristat Acetat und Luminol auf die Chemilumineszenz phagocytierender Granulocyten diskutiert.
Die angegebene Methode ermöglicht Aussagen über die Kapazität und den zeitlichen Ablauf der Stimulierbarkeit des oxidativen Metabolismus phagocytierender neutrophiler Granulocyten.
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Die Untersuchungen wurden mit Sachhilfen der Deutschen Forschungsgemeinschaft durchgeführt (Es 26/4)
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Eschenbach, C. Luminol-Chemilumineszenz phagocytierender neutrophiler Granulocyten. Klin Wochenschr 60, 357–364 (1982). https://doi.org/10.1007/BF01721626
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DOI: https://doi.org/10.1007/BF01721626