Polarographic micromethods for the rapid assay of phagocytosis-connected oxygen consumption by leukocytes in diluted peripheral blood.
Simple and quantitative micromethods were devised for the detection of phagocytosis-connected oxygen consumption by leukocytes in diluted peripheral blood. A 20 microliter volume of heparinized peripheral blood was diluted with 180 microliter of HEPES-saline buffer and treated with CO; the mixture was put in an assay well (CO micromethod) or equilibrated with air by stirring it in the well for 2 min (air micromethod). The well was revised to be suitable for the micromethods. Oxygen concentration of the blood was monitored by a Clark-type electrode fitted to the well. The phagocytosis-connected oxygen consumption by leukocytes was induced by the addition of opsonized zymosan to the blood. Both methods were ascertained to give essentially the same activity. The mean specific activity (nmol/min/10(5) cells) of oxygen consumption by phagocytizing leukocytes was 0.65 +/- 0.20 in blood from healthy adults (n = 21). No increase in oxygen consumption by leukocytes was induced in blood from patients with CGD during phagocytosis. The activities of blood from mothers of boys with CGD were 0.29 and 0.74, the latter of which did not necessarily indicate a carrier status.
D006105:Granulomatous Disease, Chronic / Q000097:blood* / Q000503:physiopathology; D006801:Humans; D007962:Leukocytes / Q000502:physiology*; D008832:Microchemistry / Q000295:instrumentation; D009895:Opsonin Proteins; D010101:Oxygen Consumption; D010587:Phagocytosis; D011048:Polarography / Q000295:instrumentation; D015054:Zymosan
D009895:Opsonin Proteins; D015054:Zymosan