Abstract
Human peripheral blood neutrophils obtained from healthy adults were examined in vitro. We assessed the effects of sequential stepwise increases in the concentration of the chemotactic dipeptide N-formyl-l-methionyl-l-phenylalanine (f-Met-Phe) on neutrophil attachment to serum-coated glass, detachment from serum-coated glass and the distribution on the cell surface of binding sites for albumin-coated latex beads. The initial exposure to f-Met-Phe resulted in increased adhesiveness and binding of latex beads in a random pattern over the cell surface. The second exposure to f-Met-Phe resulted in decreased adherence, detachment of neutrophils from serum-coated glass, and movement of binding sites for latex beads to the uropod. Enhanced adhesiveness and redistribution of binding sites were blocked by 0.1 mM N-alpha-p-tosyl-l-lysine chloromethyl ketone, a concentration that did not reduce the change in cellular shape caused by f-Met-Phe. Cytochalasin B (5 mug/ml) blocked the redistribution of binding sites as well as the change in shape. The third exposure to f-Met-Phe was given along with the latex beads. The stimulus was stopped after 2 min by fixing cells in suspension with glutaraldehyde. If the third exposure was at a concentration higher than the second, the beads were bound in the region of the lamellipodia in 70% of the cells. If lower, binding to the lamellipodia was found in a significantly smaller proportion of cells (13%). The results support the concept that neutrophils develop a polarized distribution of f-Met-Phe-induced adhesion sites in response to increasing concentrations of f-Met-Phe, and these sites flow from the region of the lamellipodia to the uropod.
MeSH Terms
Adult
Binding Sites
Cell Adhesion/drug effects
Chemotactic Factors/pharmacology
Cytochalasin B/pharmacology
Dipeptides/pharmacology
Female
Humans
Male
Microscopy, Electron, Scanning
N-Formylmethionine/analogs & derivatives,pharmacology
Neutrophils/cytology,drug effects,physiology
Serum Albumin
Tosyllysine Chloromethyl Ketone/pharmacology
Chemicals
Chemotactic Factors
Dipeptides
Serum Albumin
Tosyllysine Chloromethyl Ketone
N-formylmethionylphenylalanine
Cytochalasin B
N-Formylmethionine
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Smith C W
Hollers J C
References (16)
16 references, click to expand
-
The deactivation of rabbit neutrophils by chemotactic factor and the nature of the activatable esterase.
J Exp Med. 1968 Apr 1;127(4):693-709
PMID: 5642465
-
Alteration of the cell periphery during granulocyte maturation: relationship to cell function.
Blood. 1972 Mar;39(3):301-16
PMID: 5059645
-
Factors affecting the redistribution of surface-bound concanavalin A on human polymorphonuclear leukocytes.
J Cell Biol. 1974 Aug;62(2):351-65
PMID: 4426911
-
Active site chemotactic factors and the regulation of the human neutrophil chemotactic response.
Antibiot Chemother (1971). 1974;19:218-32
PMID: 4618739
-
A visual analysis of chemotactic and chemokinetic locomotion of human neutrophil leucocytes. Use of a new chemotaxis assay with Candida albicans as gradient source.
Exp Cell Res. 1978 Jan;111(1):191-203
PMID: 340239
-
Chemiluminescence of phagocytic cells caused by N-formylmethionyl peptides.
J Exp Med. 1978 Jan 1;147(1):182-95
PMID: 627835
-
Chemotaxis by polymorphonuclear leukocytes.
J Cell Biol. 1978 May;77(2):269-87
PMID: 649652
-
Analogous ultrastructure and surface properties during capping and phagocytosis in leukocytes.
J Cell Biol. 1978 Jun;77(3):789-804
PMID: 567226
-
Intracellular control of human neutrophil secretion. I. C5a-induced stimulus-specific desensitization and the effects of cytochalasin B.
J Immunol. 1978 Sep;121(3):851-5
PMID: 211165
-
Effects of albumin on the phagocytosis of polystyrene spherules by rabbit polymorphonuclear leucocytes.
J Cell Physiol. 1978 Oct;97(1):29-36
PMID: 568628
-
Chemotactic deactivation of human neutrophils: evidence for nonspecific and specific components.
Infect Immun. 1978 Nov;22(2):441-4
PMID: 730365
-
Chemotactic deactivation of human neutrophils: possible relationship to stimulation of oxidative metabolism.
Infect Immun. 1979 Feb;23(2):282-6
PMID: 422242
-
Motility and adhesiveness in human neutrophils. Effects of chemotactic factors.
J Clin Invest. 1979 Feb;63(2):221-9
PMID: 372238
-
Desensitization and deactivation of the secretory responsiveness of rabbit neutrophils induced by the chemotactic peptide, formyl-methionyl-leucyl-phenylalanine.
J Reticuloendothel Soc. 1979 Feb;25(2):139-50
PMID: 439096
-
Modulating influence of chemotactic factor-induced cell adhesiveness on granulocyte function.
J Clin Invest. 1979 Jul;64(1):8-16
PMID: 447862
-
Ability of polymorphonuclear leukocytes to orient in gradients of chemotactic factors.
J Cell Biol. 1977 Nov;75(2 Pt 1):606-16
PMID: 264125