Src family kinase-mediated vesicle trafficking is critical for neutrophil basement membrane penetration

Leukocyte recruitment into inflamed tissue is highly dependent on the activation and binding of integrins to their respective ligands, followed by the induction of various signaling events within the cell referred to as outside-in signaling. Src family kinases are the central players in the outside-in signaling process, assigning them a critical role for proper immune cell function. Our study investigated the role of SFKs on neutrophil recruitment in vivo using Hck-/- Fgr-/- Lyn-/- mice, which lack SFKs expressed in neutrophils. We show that loss of SFKs strongly reduces neutrophil adhesion and post-arrest modifications in a shear force dependent manner. Additionally, we found that in the absence of SFKs, neutrophils display impaired Rab27a-dependent surface mobilization of neutrophil elastase, VLA3 and VLA6 containing vesicles. This results in a defect in neutrophil vascular basement membrane penetration and thus strongly impaired extravasation. Taken together, we demonstrate a role of SFKs in neutrophil post-arrest modifications and extravasation during acute inflammation. These findings may support the current efforts to use SFK-inhibitors in inflammatory diseases with unwanted neutrophil recruitment.