A rapid technique for the isolation of highly purified, functionally intact bovine neutrophilic granulocytes.

A new method for the isolation of bovine neutrophilic granulocytes from peripheral blood based on centrifugation in a discontinuous metrizamide gradient has been developed. The procedure is rapid, taking only about 2 h, and gives highly purified (greater than 90%) neutrophils in a high yield (approximately 85%). The function of the cells, as measured by chemiluminescence and migration assays, is not significantly influenced by the isolation procedure. Eosinophils can also be isolated by a slight variation of the method. Initial applications of the separation procedure indicate its usefulness in clinical studies of bovine neutrophil function. A variation between individuals in the function of the cells was thus demonstrated. Moreover, the chemiluminescence of neutrophils from infected animals was found to be greatly increased, and neutrophil migration was shown to be stimulated by in vivo ACTH treatment.