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Heterogeneity in the circulating neutrophil pool: studies on subpopulations separated by continuous flow electrophoresis
Author(s) -
Eggleton Paul,
Fisher D.,
Crawford Neville
Publication year - 1992
Publication title -
journal of leukocyte biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.819
H-Index - 191
eISSN - 1938-3673
pISSN - 0741-5400
DOI - 10.1002/jlb.51.6.617
Subject(s) - chemotaxis , superoxide , biology , respiratory burst , phagocytosis , n formylmethionine leucyl phenylalanine , microbiology and biotechnology , in vivo , stimulation , neutrophile , biophysics , in vitro , biochemistry , immunology , endocrinology , receptor , enzyme
Isolated blood neutrophils from healthy individuals have been separated by continuous flow electrophoresis (CFE) as a Gaussian‐shaped profile extending over 12–15 fractions, on the basis of differences in cell surface electrical charge. The fractions were pooled into three or four subpopulations; the mean electrophoretic mobilities of the least and most electronegative cells were 0.96 and 1.22 μm/sec/V/cm, respectively. Each pool of neutrophils was analyzed for functional and biochemical differences. Expression of respiratory burst in terms of the rate of superoxide production by the least and most electronegative cells to fixed concentrations of N ‐formyl‐methionyl‐leucyl‐phenylalanine (fMLP, 10 −7 M) or phorbol myristate acetate (PMA, 1 μg/ml) revealed that the least electronegative cells generated superoxide anion (O 2 − ) at approximately twice the rate of the most electronegative cells. However, when lower concentrations of fMLP were used (1–5 × 10 −9 M), the most electronegative cells were most active. The least electronegative cells were also the most active in terms of phagocytosis and chemotaxis. In accordance with these differences in motile function, the basal F‐actin content of the least electronegative cell pool was greater than the F‐actin levels found in the most electronegative cells and remained so upon stimulation with fMLP. Such neutrophil heterogeneity as detected by CFE may be important in selective marginatum and recruitment of cells to inflammatory foci and sites of infection and may in part represent subsets of cells within the circulation that are primed in vivo to respond to inflammatory stimuli.