Functional Differentiation of Normal Human Neutrophils

In the past differentiation of human neutrophils has been defined by morphology, cytochemistry, or surface markers. In our experiments we have sequenced the various events that occur during the functional differentiation of the normal human neutrophil and have also examined some of the functional pr...

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Bibliographic Details
Published in:Blood 1987-03, Vol.69 (3), p.937-944
Main Authors: Glasser, Lewis, Fiederlein, Roger L.
Format: Article
Language:English
Subjects:
Bacteriolysis
Biological and medical sciences
Cell Differentiation
Cell Separation
Chemotaxis, Leukocyte
Cytotoxicity, Immunologic
Fundamental and applied biological sciences. Psychology
Humans
Neutrophils - classification
Neutrophils - cytology
Neutrophils - physiology
Nitroblue Tetrazolium - metabolism
Oxidation-Reduction
Oxygen - physiology
Phagocytosis
Receptors, Complement - analysis
Receptors, Fc - analysis
Vertebrates: blood, hematopoietic organs, reticuloendothelial system
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Summary:In the past differentiation of human neutrophils has been defined by morphology, cytochemistry, or surface markers. In our experiments we have sequenced the various events that occur during the functional differentiation of the normal human neutrophil and have also examined some of the functional properties in relationship to surface markers and biochemical events. Granulocytes were obtained from the bone marrow and blood of hematologically normal individuals. Cells were separated into different stages of maturation by their physical properties using counterflow centrifugal elutriation and density gradient separation. Three cell fractions were obtained that were enriched for either immature myeloid cells, band neutrophils, or segmented neutrophils. Since the enriched fractions were not entirely pure, methodologies for functional assays were chosen that allowed cytologic evaluation of the functional capacity of each cell type. The criteria used to classify the stages of differentiation included both morphology by light microscopy and DNA labeling with tritiated thymidine. Various neutrophilic properties were studied: Fc receptors, complement receptors (CR1, CR3), phagocytosis of both live and dead opsonized Staphylococcus aureus, microbial killing of S aureus, NBT dye reduction after cellular stimulation with endotoxin, and chemotaxis. Our results indicate that the functional properties of the neutrophil appear in a distinct order. The sequence for the functional differentiation of the human neutrophil appears to be the following: Fc receptors → immune phagocytosis → complement receptors → oxygen-independent microbial killing → oxy-gen-dependent microbial killing → chemotaxis. © 1987 by Grune & Stratton, Inc.
ISSN:0006-4971
1528-0020
DOI:10.1182/blood.V69.3.937.937