Cytoplasts made from human blood polymorphonuclear leukocytes with or without heat: preservation of both motile function and respiratory burst oxidase activity.

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RESUMO

Anucleate fragments (cytoplasts) from polymorphonuclear leukocytes (PMN) are simplified systems that can be used to elucidate specific pathways by which cell function is altered. PMN cytoplasts in current use are defective either in activatable respiratory burst oxidase activity or in motile function. By centrifugation of PMN on discontinuous gradients of Ficoll without cytochalasin B, we have created granule-poor cytoplasts in which both these capacities are preserved. Specifically, they generate superoxide anion (O2-.) and reduce nitroblue tetrazolium dye on appropriate stimulation; they respond chemotactically to erythrocytes destroyed by laser microirradiation or to the specific chemoattractants fMet-Leu-Phe (10 nM) and C5a (zymosan-activated serum); and they ingest and kill staphylococci. We can improve the yield of these fragments progressively by preheating (45 degrees C) the cells in suspension for increasing periods of time, but those treatments are attended by a decreasing percentage of cytoplasts with activatable oxidase activity, and a progressive inability of the cytoplasts to ingest and to kill staphylococci. These easily made and multipotent cytoplasts readily lend themselves to studies of PMN physiology.

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