The migration of neutrophils into inflamed tissues is a fundamental component of innate immunity. A decisive step in this process is the polarized migration of blood neutrophils through endothelial cells (ECs) lining the venular lumen (transendothelial migration (TEM)) in a luminal-to-abluminal direction. By real-time confocal imaging, we found that neutrophils had disrupted polarized TEM ('hesitant' and 'reverse') in vivo. We noted these events in inflammation after ischemia-reperfusion injury, characterized by lower expression of junctional adhesion molecule C (JAM-C) at EC junctions, and they were enhanced by blockade or genetic deletion of JAM-C in ECs. Our results identify JAM-C as a key regulator of polarized neutrophil TEM in vivo and suggest that reverse TEM of neutrophils can contribute to the dissemination of systemic inflammation.
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We thank E. Dejana (FIRC Institute of Molecular Oncology) for mAb BV11 to JAM-A; S.J. Weiss for critical reading of the manuscript; R. Yadav for advice on surgical procedures; and H.M. McGetterick for contributions to analysis of rTEM neutrophils generated in vitro. Supported by the Wellcome Trust (081172/Z/06/Z to S.N.), the UK National Institute for Health Research (for the translational research portfolio of Barts and the London Cardiovascular Biomedical Research Unit, to which the work of S.N. contributes), the Swiss National Science Foundation (310000-122423, 310000-109402 and CR32I3_129987 to P.M. and 310030-120184 for B.A.I.), the Juvenile Diabetes Research Foundation (40-2011-11 to P.M.), the European Union (BETAIMAGE 222980; IMIDIA and C2008-T7 to P.M.; and a Marie Curie Fellowship (FP7-PEOPLE-2009-IEF-252091) to M.B.), the Intramural Research Program of the US National Institutes of Health (T.C.), the National Cancer Institute (T.C.) and Deutsche Forschungsgemeinschaft (T.C.).
The authors declare no competing financial interests.
Supplementary Figures 1–6, Supplementary Methods and Supplementary Results (PDF 752 kb)
Development of an inflammatory response in an IL-1β-stimulated tissue. (WMV 1553 kb)
Migration of leukocytes in a paracellular mode. (WMV 1172 kb)
Paracellular transmigration and pore formation (WMV 1058 kb)
Transcellular TEM and pore formation (WMV 1067 kb)
Hesitant TEM as induced by I-R injury (example 1). (WMV 1373 kb)
Hesitant TEM as induced by I-R injury (example 2). (WMV 1259 kb)
Hesitant TEM as induced by I-R injury (example 3). (WMV 917 kb)
Reverse TEM as induced by I-R injury. (WMV 1847 kb)
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Woodfin, A., Voisin, M., Beyrau, M. et al. The junctional adhesion molecule JAM-C regulates polarized transendothelial migration of neutrophils in vivo. Nat Immunol 12, 761–769 (2011). https://doi.org/10.1038/ni.2062
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