Abstract
At sites of inflammation, ligation of leukocyte integrins is critical for the activation of cellular effector functions required for host defense. However, the signaling pathways linking integrin ligation to cellular responses are poorly understood. Here we show that integrin signaling in neutrophils and macrophages requires adaptors containing immunoreceptor tyrosine-based activation motifs (ITAMs). Neutrophils and macrophages lacking two ITAM-containing adaptor proteins, DAP12 and FcRγ, were defective in integrin-mediated responses. Activation of the tyrosine kinase Syk by integrins required that DAP12 and FcRγ were first phosphorylated by Src family kinases. Retroviral transduction of neutrophils and macrophages with wild-type and mutant Syk or DAP12 demonstrated that the Src homology 2 domains of Syk and the ITAM of DAP12 were required for integrin signaling. Our data show that integrin signaling for the activation of cellular responses in neutrophils and macrophages proceeds by an immunoreceptor-like mechanism.
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Acknowledgements
We thank Y. Refaeli for the pMIG-W vector; K. Makara and Á. Mikesy for mouse colony management; M. Kovács for help with experiments; and A. Erdei for access to equipment. Syk+/− mice and anti-Syk were from V. Tybulewicz (National Institute for Medical Research); Itgb2−/− (CD18-knockout) mice were from A. Beaudet (Baylor College of Medicine); anti-DAP12 was from T. Takai (Tohoku University); and the GST-Syk-(SH2)2 fusion protein was from A. DeFranco (University of California, San Francisco). Supported by the US National Institutes of Health (TW006831 to C.A.L. and A.M.; AI065150 and AI065495 to C.A.L.; and AI068129 to L.L.L.), the Hungarian National Scientific Research Fund (T046409 to A. M.), the Wellcome Trust (A.M.), the European Molecular Biology Organization–Howard Hughes Medical Institute (A.M.), the Hungarian Academy of Sciences (Bolyai Research Fellowship to A.M.) and the American Cancer Society Research (L.L.L.).
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A.M. and C.A.L. initiated the study; A.M., C.L.A., Z.J. and C.A.L. designed and did the experiments; A.M. supervised the work in Budapest; Y.H. did fetal liver stem cell transfers and maintained mouse colonies; L.L.L. provided intellectual guidance and manuscript editing; and A.M., C.L.A. and C.A.L. wrote the manuscript.
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Supplementary information
Supplementary Fig. 1
Normal integrin and Gr1 expression on neutrophils lacking DAP12 and FcRγ. (PDF 57 kb)
Supplementary Fig. 2
ITAM-like signaling in human neutrophils. (PDF 107 kb)
Supplementary Fig. 3
Lack of the FcRγ-chain associated kinase activity in Syk-KO neutrophils. (PDF 94 kb)
Supplementary Fig. 4
Model of integrin signaling in neutrophils and macrophages. (PDF 50 kb)
Supplementary Fig. 5
Normal expression of β2-integrins on macrophages lacking DAP12 and FcRγ. (PDF 46 kb)
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Mócsai, A., Abram, C., Jakus, Z. et al. Integrin signaling in neutrophils and macrophages uses adaptors containing immunoreceptor tyrosine-based activation motifs. Nat Immunol 7, 1326–1333 (2006). https://doi.org/10.1038/ni1407
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DOI: https://doi.org/10.1038/ni1407
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