MODULATION OF MULTIPLE NEUTROPHIL FUNCTIONS BY PREPARATIVE METHODS OR TRACE CONCENTRATIONS OF BACTERIAL LIPOPOLYSACCHARIDE

Google Scholar
  • 1 January 1985
    • journal article
    • research article
    • Vol. 119  (1) , 101-110
Abstract
Human neutrophils were isolated from peripheral blood by four methods: Ficoll-Hypaque gradients and erythrocyte lysis, plasma-Percoll gradients, a lipopolysaccharide (LPS)-free method yielding 85% neutrophils, and by centrifugation of cells prepared by method 3 through a plasma-Percoll gradient to produce pure neutrophils. The use of the Ficoll-Hypaque method resulted in spontaneous change of cell shape, enhanced for Met-Leu-Phe (FMLP)-stimulated release of superoxide anion, increased-release of lysosomal enzymes upon subsequent FMLP stimulation, and reduced chemotactic responsiveness, by comparison with the other methods. These effects were not due to erythrocyte lysis by NH4C1 but were reproduced by exposure of neutrophils prepared by the LPS-free method or the use of plasma-Percoll gradients to 10-100 ng/ml LPS. Neutrophil change of shape and stimulated O2- production were particularly sensitive markers of these effects. The effects of trace concentrations of LPS in the modulation of neutrophil function may have relevance to the pathophysiology of endotoxemia and its resultant tissue injury.