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Analysis of Human Monocyte Chemoattractant Binding by Flow Cytometry
Author(s) -
Leonard Edward J.,
Noer Kathleen,
Skeel Alison
Publication year - 1985
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.38.3.403
Subject(s) - flow cytometry , monocyte , chemotaxis , biology , receptor , peptide , peripheral blood mononuclear cell , peripheral blood , cytometry , immunology , microbiology and biotechnology , biochemistry , in vitro
The chemotactic peptide fMet‐Leu‐Phe‐Lys is a potent chemoattractant for human blood monocytes. However, only one‐third of the monocytes respond. To determine whether or not lack of response reflected absence of attractant receptors, we equilibrated peripheral blood mononuclear cells with fMet‐Leu‐Phe‐Lys‐FITC and analyzed binding by flow cytometry. The fluoresceinated peptide bound rapidly at 0°, and the amount bound approached saturation with increasing concentration. The percentage of blood monocytes that bound the peptide was 60 ± 8 (SEM for seven experiments). In contrast, only 36 ± 3% (SEM for 16 experiments) of monocytes responded to the attractant by directed migration. It follows that, among the 64 nonmigrating monocytes per 100 total monocytes, approximately 40, or two‐thirds of them, fail to bind attractant; the remaining one‐third bind attractant but do not respond with directed movement.