Assessment of Arf6 Deletion in PLB-985 Differentiated in Neutrophil-Like Cells and in Mouse Neutrophils: Impact on Adhesion and Migration

• Published in: Mediators of inflammation Vol. 2020; no. 2020; pp. 1 - 13
• Main Authors: Bourgoin, Sylvain G., Kanaho, Yasunori, Funakoshi, Yuji, Hongu, Tsunaki, Rollet-Labelle, Emmanuelle, Davis, Lynn, Gamara, Jouda, Aoudjit, Fawzi
• Format: Journal Article
• Language: English
• Published: Cairo, Egypt Hindawi Publishing Corporation 2020; Hindawi; John Wiley & Sons, Inc; Hindawi Limited
• Subjects: ADP-Ribosylation Factors - genetics; ADP-Ribosylation Factors - metabolism; Animals; Apoptosis; Apoptosis - genetics; Apoptosis - physiology; Blood; Blood Cell Count; Blotting, Western; Bone marrow; CD11a antigen; CD11b antigen; Cell adhesion & migration; Cell differentiation; Cell Line; Cell surface; Cells, Cultured; Cloning; Endothelium; Ethylenediaminetetraacetic acid; Extracellular Matrix Proteins - genetics; Extracellular Matrix Proteins - metabolism; Fibrin; Fibrinogen; Fibronectin; Fibronectins; Flow Cytometry; Formyl peptides; Genetic Vectors - genetics; Health aspects; Humans; Immunodeficiency; Inflammation; Integrin beta Chains - genetics; Integrin beta Chains - metabolism; Integrins; Intercellular adhesion molecule 1; Kinases; Leukocyte migration; Leukocytes (neutrophilic); Ligands; Lipopolysaccharides; Lymphocyte Function-Associated Antigen-1 - genetics; Lymphocyte Function-Associated Antigen-1 - metabolism; Macrophage-1 Antigen - genetics; Macrophage-1 Antigen - metabolism; Mice; Mice, Knockout; Neutrophils; Neutrophils - cytology; Neutrophils - metabolism; Peptides; Proteins; Reverse Transcriptase Polymerase Chain Reaction; RNA, Small Interfering - genetics; RNA, Small Interfering - metabolism; Canada; United States > US
• ISSN: 0962-9351; 1466-1861
• DOI: 10.1155/2020/2713074

Chemoattractant sensing, adhesiveness, and migration are critical events underlying the recruitment of neutrophils (PMNs) to sites of inflammation or infection. Defects in leukocyte adhesion or migration result in immunodeficiency disorders characterized by recurrent infections. In this study, we evaluated the role of Arf6 on PMN adhesion in vitro and on migration to inflammatory sites using PMN-Arf6 conditional knockout (cKO) mice. In PMN-like PLB-985 silenced for Arf6 fMLP-mediated adhesion to the β2 integrin ligands, ICAM-1 and fibrinogen or the β1/β2 integrin ligand fibronectin was significantly reduced. Furthermore, overexpression of wild-type Arf6 promoted basal and fMLP-induced adhesion to immobilized integrin ligands, while overexpression of the dominant-negative Arf6 has the opposite effects. Using the Elane-Cre deleting mouse strains, we report that the level of Arf6 deletion in inflammatory PMNs isolated from the dorsal air pouches was stronger when compared to naïve cells isolated from the bone marrow. In PMN-Arf6 cKO mice, the recruitment of PMNs into the dorsal air pouch injected with LPS or the chemoattractant fMLP was significantly diminished. Impaired cell migration correlated with reduced cell surface expression of CD11a and CD11b in Arf6 cKO PMNs. Our results highlight that Arf6 regulates the activity and possibly the recycling of PMN integrins, and this compromises PMN migration to inflammatory sites.