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Isolation and phagocytic properties of neutrophils and other phagocytes from nonmastitic bovine milk.

A technique for the separation of neutrophils from macrophages-epithelial cells in samples of nonmastitic bovine milk with low cell counts has been developed. The procedure is based on centrifugation in a discontinuous metrizamide gradient and is rapid, taking less than 40 min. The recovery of the neutrophils is about 30% and their viability about 90%. The isolated neutrophils showed an appreciable unstimulated luminol- and lucigenin-dependent chemiluminescence, which was due to NADPH oxidase rather than to xanthine oxidase. The neutrophils had a higher rate of ingestion of C3-opsonized particles than macrophages-epithelial cells, whereas no significant differences in phagocytosis of IgG-opsonized yeast or unopsonized yeast were detected between the two cell populations. The macrophages-epithelial cells produced no luminol-dependent chemiluminescence and induced considerably lower activity in the lucigenin-dependent system than neutrophils, indicating that these cells contain no myeloperoxidase. Analyses of the activity of the neutrophils in response to C3-opsonized yeast particles showed that the luminol-dependent chemiluminescence of cells isolated from residual milk increased significantly over the lactation period. Moreover, a tendency to a higher phagocytosis and chemiluminescence of neutrophils isolated from residual milk than from stripping milk was indicated.

Animals↗

Inhibitory effect of antibiotic cerulenin on the respiratory burst in phagocytes. I. Effects of cerulenin on active oxygen-generation and lipid metabolism in phagocytes.

The antibiotic cerulenin, a known inhibitor of fatty acid and sterol synthesis, inhibited bactericidal activity of mouse peritoneal macrophages and chemiluminescence (CL) response upon phagocytosis. The antibiotic also inhibited the CL response of human neutrophils upon exposure to various stimuli such as chemotactic peptide N-formylmethionylleucylphenylalanine (fMLP), calcium ionophore A23187 and Staphylococcal delta toxin. The loss of CL response of both types of cells was observed only after incubation of the cells with cerulenin for certain periods. The results of radioactive precursor incorporation suggest that lipid metabolism blocked by cerulenin affected in turn signal transduction across the cell membrane and inhibited CL production in these cells.

Animals↗