title: Differentiated HL-60 cells are a valid model system for the analysis of human neutrophil migration and chemotaxis.
abstract: We have carried out a detailed comparison of the motile properties of differentiated HL-60 cells and human peripheral blood neutrophils. We compared the effects of chemotactic stimuli and of inhibitors of signalling proteins on morphology, chemokinesis and chemotaxis of neutrophils and differentiated HL-60 cells using videomicroscopy and a filter assay for chemotaxis. We also assessed expression of signalling and cytoskeletal proteins using Western blotting. Chemotactic peptide induced a front-tail polarity in HL-60 cells comparable to that of neutrophils. Chemokinetic and chemotactic responses to chemotactic peptide were also very similar for both cell types, concerning mean speed of migration, the fraction of migrated cells and the concentration of stimulus optimal for activation. The cytokine interleukin-8 was in contrast clearly less effective in activating motile responses of differentiated HL-60 cells as compared to neutrophils. An important functional role of Rho-activated kinases and phosphatidylinositol 3-kinase in motile responses of HL-60 cells, consistent with their upregulation during differentiation, could be confirmed using inhibitors with specificity for the corresponding enzymes. The only difference observed here between HL-60 cells and neutrophils concerned the differential effects of a protein kinase C inhibitor.In summary, the results presented here show that differentiated HL-60 cells, stimulated with chemotactic peptide, are a valid model system to study molecular mechanisms of neutrophil emigration.
authors.affiliation: Department of Pathology, University of Bern, Murtenstr. 31, CH-3010 Bern, Switzerland.
authors.forename: Anna Barbara
authors.initials: AB
authors.lastname: Hauert
authors.forename: Sibylla
authors.initials: S
authors.lastname: Martinelli
authors.forename: Camilla
authors.initials: C
authors.lastname: Marone
authors.forename: Verena
authors.initials: V
authors.lastname: Niggli
mesh_terms: D000577:Amides / Q000494:pharmacology; D002454:Cell Differentiation; D002465:Cell Movement / Q000187:drug effects / Q000502:physiology*; D016764:Cell Polarity; D048429:Cell Size; D002634:Chemotaxis, Leukocyte / Q000187:drug effects / Q000502:physiology*; D003598:Cytoskeletal Proteins / Q000378:metabolism; D019206:GTP-Binding Protein alpha Subunits, Gi-Go / Q000378:metabolism; D018922:HL-60 Cells / Q000166:cytology* / Q000187:drug effects / Q000502:physiology*; D006801:Humans; D066298:In Vitro Techniques; D016209:Interleukin-8 / Q000494:pharmacology; D047908:Intracellular Signaling Peptides and Proteins; D008954:Models, Biological; D009504:Neutrophils / Q000166:cytology* / Q000187:drug effects / Q000502:physiology*; D009842:Oligopeptides / Q000494:pharmacology; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism; D011493:Protein Kinase C / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors; D011725:Pyridines / Q000494:pharmacology; D015398:Signal Transduction; D054460:rho-Associated Kinases
chemical_list: D000577:Amides; D003598:Cytoskeletal Proteins; D019206:GTP-Binding Protein alpha Subunits, Gi-Go; D016209:Interleukin-8; D047908:Intracellular Signaling Peptides and Proteins; D009842:Oligopeptides; D019869:Phosphatidylinositol 3-Kinases; D010744:Phosphoric Monoester Hydrolases; D011493:Protein Kinase C; D017346:Protein Serine-Threonine Kinases; D011725:Pyridines; D054460:rho-Associated Kinases; C108830:Y 27632; C021642:F-chemotactic peptide