Determination of Chemiluminescence Associated with Phagocytosis by Human Polymorphonuclear Leukocytes
McKean, Theodore J.
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Polymorphonuclear leukocytes (PMNLs) are a vital component of many inflammatory responses which provide a protective mechanism against many pathogens. But as the oxidants that PMNLs produce are damaging to healthy tissue, the relative benefits of a response which may help to contain affected tissues, must be compared to the potential for causing further damage (to surrounding tissue). For this reason, an assay capable of revealing PMNL oxygenation activity of intact cells is necessary. A chemiluminescence assay was developed to detect damaging oxidants that activated PMNLs produce, and the modulatory effect of various compounds were investigated. The cells were activated with polystyrene microspheres or yeast particles in mixtures containing either isolated PMNLs or whole blood samples. The anticoagulants EDTA and sodium heparin were found to inhibit oxygenation activity of PMNLs. Inhibition during the early stages of PMNL activation was found to be reversible if calcium ions were introduced. It was proposed that calcium is involved in the binding of the stimulant particle to the membrane of the PMNL. The effect of two experimental Upjohn drugs belonging to a family called "Lazaroid" on the oxygenation activity of PMNLs was found to be insignificant, although these drugs did reveal an inhibitory effect in two homogeneous enzyme systems that generated the same oxidants that PMNLs produce.