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Src kinases and neutrophil extravasation
dependent LN degradation creates peptides that are chemoattractant for neutrophils.33 In line with these find- ings, we were able to show diminished LN degradation by SFK-ko neutrophils in vitro using spinning disk confocal microscopy. Surprisingly, we observed that SFK-ko neu- trophils, which are unable to degrade the LN barrier them- selves, arrange around wildtype neutrophils, strongly sup- porting the hypothesis of the chemoattractant function of LN-fragments. Although our data seem to partly contrast with the study by Kovács et al., in which no intrinsic neu- trophil migration defect was observed in the genetic absence of SFK,15 we propose that this is due to the differ- ent models used. It is also likely that the rescued extrava- sation into the synovial joint in their mixed chimeric mice (50% SFK-ko, 50% wildtype neutrophils) is in part due to restored NE or protease secretion and subsequent BM degradation by wildtype neutrophils.
Taken together, we have demonstrated a critical role for SFK in various steps along the neutrophil adhesion cas- cade during acute inflammation. While its effect on adhe- sion strengthening is the direct consequence of its promi- nent role directly downstream of integrin ligation and sig-
naling, the regulation of vesicle trafficking during neu- trophil extravasation is unexpected and provides new insights on the extravasation process through the vascular BM, supporting the concept of neutrophil-mediated vas- cular BM digestion along this route.
Acknowledgments
The authors would like to thank Susanne Bierschenk and Nadine Schmidt for their excellent technical assistance, as well as Andreas Thomae and Steffen Dietzel (Core Facility BioImaging, BioMedical Center, Ludwig-Maximilians-Universität München) for their help with confocal and multi-photon microscopy. The authors thank Thomas Graf for providing the Lyz2GFP mice.
Funding
This work was supported by grants from the European Community’s Seventh Framework Programme (FP7-2007- 2013) under grant agreement HEALTH-F4-2011-282095 (TARKINAID to MS and AM), Deutsche Forschungsgemeinschaft (SFB 914, project B01 [MS], Z03 [MS]) and the Else Kröner-Fresenius-Stiftung (grant 2015_A68 [IR]).
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