title: HS1 deficiency impairs neutrophil recruitment in vivo and activation of the small GTPases Rac1 and Rap1.
abstract: Neutrophil extravasation is a critical step of the innate immune system's response to inflammation. This multistep process is tightly regulated by adhesion and signaling molecules in the endothelium and neutrophils. Activation of the β2integrin LFA-1 is critical for adhesion of leukocytes to postcapillary venules. This step requires coordinated activation of signaling pathways in chemokine-stimulated neutrophils, including GTPase activation and cytoskeletal remodeling, leading to conformational changes in LFA-1. Hematopoietic cell-specific lyn substrate 1 (HS1) is a cortactin-related and leukocyte-specific actin-binding protein (ABP) that regulates several processes in various immune cells. It has been shown in vitro that HS1 is important for neutrophil chemotaxis and transendothelial migration of NK cells, but its role in neutrophil extravasation in vivo has not been investigated yet. Intravital microscopy of CXCL1-stimulated cremaster venules revealed an increased rolling velocity and reduced neutrophil adhesion and transmigration in HS1 knockout (KO) mice. CXCL1-induced rapid neutrophil arrest in vivo and adhesion under flow conditions in vitro were also reduced significantly. Whereas random motility of neutrophils was unaffected, chemotaxis toward a CXCL1 gradient was reduced in the absence of HS1. Further analysis of the underlying mechanisms demonstrated that HS1 controls CXCL1-induced activation of the small GTPases Ras-related C3 botulinum toxin substrate 1 (Rac1) and Ras-related protein 1 (Rap1), thus supporting LFA-1-mediated neutrophil adhesion. Importantly, with the use of Rac1 KO neutrophils, we could show that Rac1 acts upstream of Rap1. Our results establish HS1 as an important regulator of proper Rac1 and Rap1 activation and neutrophil extravasation.
authors.affiliation: Department for Vascular Cell Biology, Max Planck Institute for Molecular Biomedicine, Münster, Germany; and.
authors.forename: Joanna
authors.initials: J
authors.lastname: Latasiewicz
authors.affiliation: Department for Vascular Cell Biology, Max Planck Institute for Molecular Biomedicine, Münster, Germany; and.
authors.forename: Annette
authors.initials: A
authors.lastname: Artz
authors.affiliation: Department for Vascular Cell Biology, Max Planck Institute for Molecular Biomedicine, Münster, Germany; and.
authors.forename: Ding
authors.initials: D
authors.lastname: Jing
authors.affiliation: Department for Vascular Cell Biology, Max Planck Institute for Molecular Biomedicine, Münster, Germany; and.
authors.forename: Mariana Pacheco
authors.initials: MP
authors.lastname: Blanco
authors.affiliation: Department for Vascular Cell Biology, Max Planck Institute for Molecular Biomedicine, Münster, Germany; and.
authors.forename: Silke M
authors.initials: SM
authors.lastname: Currie
authors.affiliation: Department for Molecular Biomedicine, Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional, Mexico City, Mexico.
authors.forename: Martha Velázquez
authors.initials: MV
authors.lastname: Avila
authors.affiliation: Department for Molecular Biomedicine, Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional, Mexico City, Mexico mschnoor@cinvestav.mx.
authors.forename: Michael
authors.initials: M
authors.lastname: Schnoor
authors.affiliation: Department for Vascular Cell Biology, Max Planck Institute for Molecular Biomedicine, Münster, Germany; and vestweb@mpi-muenster.mpg.de.
authors.forename: Dietmar
authors.initials: D
authors.lastname: Vestweber
mesh_terms: D000009:Abdominal Muscles / Q000098:blood supply / Q000166:cytology / Q000276:immunology; D000818:Animals; D002448:Cell Adhesion / Q000187:drug effects; D054360:Chemokine CXCL1 / Q000235:genetics / Q000276:immunology / Q000494:pharmacology; D002633:Chemotaxis / Q000187:drug effects; D016179:Granulocyte Colony-Stimulating Factor / Q000172:deficiency / Q000235:genetics / Q000276:immunology*; D007113:Immunity, Innate; D000069416:Intravital Microscopy; D016169:Lymphocyte Function-Associated Antigen-1 / Q000235:genetics / Q000276:immunology*; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D009479:Neuropeptides / Q000235:genetics / Q000276:immunology*; D020556:Neutrophil Infiltration / Q000187:drug effects; D009504:Neutrophils / Q000187:drug effects / Q000276:immunology* / Q000473:pathology; D010538:Peritonitis / Q000235:genetics / Q000276:immunology* / Q000473:pathology; D061251:Primary Cell Culture; D020830:rac1 GTP-Binding Protein / Q000235:genetics / Q000276:immunology*; D020827:rap1 GTP-Binding Proteins / Q000235:genetics / Q000276:immunology*
chemical_list: D054360:Chemokine CXCL1; C475895:Cxcl1 protein, mouse; D016169:Lymphocyte Function-Associated Antigen-1; D009479:Neuropeptides; C484602:Rac1 protein, mouse; C524015:hematopoietic lineage cell-specific protein 1, mouse; D016179:Granulocyte Colony-Stimulating Factor; C000620839:Rap1 protein, mouse; D020830:rac1 GTP-Binding Protein; D020827:rap1 GTP-Binding Proteins
keywords: adhesion; inflammation; integrins; leukocyte extravasation; transmigration