Abstract
The effector functions of natural killer cells are regulated by activating receptors, which recognize stress-inducible ligands expressed on target cells and signal through association with signaling adaptors. Here we developed a mouse model in which a fusion of the signaling adaptor DAP10 and ubiquitin efficiently downregulated expression of the activating receptor NKG2D on the surfaces of natural killer cells. With this system, we identified coupling of the signaling pathways triggered by NKG2D and DAP10 to those initiated by the interleukin 15 receptor. We suggest that this coupling of activating receptors to other receptor systems could function more generally to regulate cell type–specific signaling events in distinct physiological contexts.
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References
Biron, C.A., Nguyen, K.B., Pien, G.C., Cousens, L.P. & Salazar-Mather, T.P. Natural killer cells in antiviral defense: function and regulation by innate cytokines. Annu. Rev. Immunol. 17, 189–220 (1999).
Smyth, M.J., Hayakawa, Y., Takeda, K. & Yagita, H. New aspects of natural-killer-cell surveillance and therapy of cancer. Nat. Rev. Cancer 2, 850–861 (2002).
Lanier, L.L. NK cell recognition. Annu. Rev. Immunol. 23, 225–274 (2005).
Raulet, D.H. Interplay of natural killer cells and their receptors with the adaptive immune response. Nat. Immunol. 5, 996–1002 (2004).
Yokoyama, W.M., Kim, S. & French, A.R. The dynamic life of natural killer cells. Annu. Rev. Immunol. 22, 405–429 (2004).
Kim, S. et al. Licensing of natural killer cells by host major histocompatibility complex class I molecules. Nature 436, 709–713 (2005).
Fernandez, N.C. et al. A subset of natural killer cells achieves self-tolerance without expressing inhibitory receptors specific for self-MHC molecules. Blood 105, 4416–4423 (2005).
Lanier, L.L. Natural killer cell receptor signaling. Curr. Opin. Immunol. 15, 308–314 (2003).
Colucci, F. et al. Natural cytotoxicity uncoupled from the Syk and ZAP-70 intracellular kinases. Nat. Immunol. 3, 288–294 (2002).
Gilfillan, S., Ho, E.L., Cella, M., Yokoyama, W.M. & Colonna, M. NKG2D recruits two distinct adaptors to trigger NK cell activation and costimulation. Nat. Immunol. 3, 1150–1155 (2002).
Bakker, A.B. et al. DAP12-deficient mice fail to develop autoimmunity due to impaired antigen priming. Immunity 13, 345–353 (2000).
Tomasello, E. et al. Combined natural killer cell and dendritic cell functional deficiency in KARAP/DAP12 loss-of-function mutant mice. Immunity 13, 355–364 (2000).
Chiesa, S. et al. Multiplicity and plasticity of natural killer cell signaling pathways. Blood 107, 2364–2372 (2006).
Wu, J. et al. An activating immunoreceptor complex formed by NKG2D and DAP10. Science 285, 730–732 (1999).
Jamieson, A.M. et al. The role of the NKG2D immunoreceptor in immune cell activation and natural killing. Immunity 17, 19–29 (2002).
Raulet, D.H. Roles of the NKG2D immunoreceptor and its ligands. Nat. Rev. Immunol. 3, 781–790 (2003).
Roth, A.F. & Davis, N.G. Ubiquitination of the PEST-like endocytosis signal of the yeast a-factor receptor. J. Biol. Chem. 275, 8143–8153 (2000).
Garrity, D., Call, M.E., Feng, J. & Wucherpfennig, K.W. The activating NKG2D receptor assembles in the membrane with two signaling dimers into a hexameric structure. Proc. Natl. Acad. Sci. USA 102, 7641–7646 (2005).
Rosen, D.B. et al. A Structural basis for the association of DAP12 with mouse, but not human, NKG2D. J. Immunol. 173, 2470–2478 (2004).
Hicke, L. & Dunn, R. Regulation of membrane protein transport by ubiquitin and ubiquitin-binding proteins. Annu. Rev. Cell Dev. Biol. 19, 141–172 (2003).
Diefenbach, A. et al. Selective associations with signaling proteins determine stimulatory versus costimulatory activity of NKG2D. Nat. Immunol. 3, 1142–1149 (2002).
Hayakawa, Y. et al. Cutting edge: tumor rejection mediated by NKG2D receptor-ligand interaction is dependent upon perforin. J. Immunol. 169, 5377–5381 (2002).
Kennedy, M.K. et al. Reversible defects in natural killer and memory CD8 T cell lineages in interleukin 15-deficient mice. J. Exp. Med. 191, 771–780 (2000).
Lodolce, J.P. et al. IL-15 receptor maintains lymphoid homeostasis by supporting lymphocyte homing and proliferation. Immunity 9, 669–676 (1998).
Matsuda, J.L. et al. Homeostasis of Vα14i NKT cells. Nat. Immunol. 3, 966–974 (2002).
Cooper, M.A. et al. In vivo evidence for a dependence on interleukin 15 for survival of natural killer cells. Blood 100, 3633–3638 (2002).
Huntington, N.D. et al. Interleukin 15-mediated survival of natural killer cells is determined by interactions among Bim, Noxa and Mcl-1. Nat. Immunol. 8, 856–863 (2007).
Meresse, B. et al. Coordinated induction by IL15 of a TCR-independent NKG2D signaling pathway converts CTL into lymphokine-activated killer cells in celiac disease. Immunity 21, 357–366 (2004).
Sechler, J.M., Barlic, J., Grivel, J.C. & Murphy, P.M. IL-15 alters expression and function of the chemokine receptor CX3CR1 in human NK cells. Cell. Immunol. 230, 99–108 (2004).
Karlhofer, F.M., Orihuela, M.M. & Yokoyama, W.M. Ly-49-independent natural killer (NK) cell specificity revealed by NK cell clones derived from p53-deficient mice. J. Exp. Med. 181, 1785–1795 (1995).
Kovanen, P.E. & Leonard, W.J. Cytokines and immunodeficiency diseases: critical roles of the γc-dependent cytokines interleukins 2, 4, 7, 9, 15, and 21, and their signaling pathways. Immunol. Rev. 202, 67–83 (2004).
Krutzik, P.O. & Nolan, G.P. Intracellular phospho-protein staining techniques for flow cytometry: monitoring single cell signaling events. Cytometry A 55, 61–70 (2003).
Zhang, X., Sun, S., Hwang, I., Tough, D.F. & Sprent, J. Potent and selective stimulation of memory-phenotype CD8+ T cells in vivo by IL-15. Immunity 8, 591–599 (1998).
Lucas, M., Schachterle, W., Oberle, K., Aichele, P. & Diefenbach, A. Dendritic cells prime natural killer cells by trans-presenting interleukin 15. Immunity 26, 503–517 (2007).
Minagawa, M. et al. Enforced expression of Bcl-2 restores the number of NK cells, but does not rescue the impaired development of NKT cells or intraepithelial lymphocytes, in IL-2/IL-15 receptor β-chain-deficient mice. J. Immunol. 169, 4153–4160 (2002).
Imada, K. et al. Stat5b is essential for natural killer cell-mediated proliferation and cytolytic activity. J. Exp. Med. 188, 2067–2074 (1998).
Sudbeck, E.A. et al. Structure-based design of specific inhibitors of Janus kinase 3 as apoptosis-inducing antileukemic agents. Clin. Cancer Res. 5, 1569–1582 (1999).
Betts, M.R. et al. Sensitive and viable identification of antigen-specific CD8+ T cells by a flow cytometric assay for degranulation. J. Immunol. Methods 281, 65–78 (2003).
Sutherland, C.L. et al. UL16-binding proteins, novel MHC class I-related proteins, bind to NKG2D and activate multiple signaling pathways in primary NK cells. J. Immunol. 168, 671–679 (2002).
Tomasello, E. & Vivier, E. KARAP/DAP12/TYROBP: three names and a multiplicity of biological functions. Eur. J. Immunol. 35, 1670–1677 (2005).
Takegahara, N. et al. Plexin-A1 and its interaction with DAP12 in immune responses and bone homeostasis. Nat. Cell Biol. 8, 615–622 (2006).
Koga, T. et al. Costimulatory signals mediated by the ITAM motif cooperate with RANKL for bone homeostasis. Nature 428, 758–763 (2004).
Mocsai, A. et al. Integrin signaling in neutrophils and macrophages uses adaptors containing immunoreceptor tyrosine-based activation motifs. Nat. Immunol. 7, 1326–1333 (2006).
Acknowledgements
We thank D. Raulet (University of California, Berkeley) for DAP10-specific antiserum and communication of unpublished results; W. Yokoyama for the KY-1 cells; S. Holley and C. Anicelli for help with mouse work; G. Tokmoulina and T. Taylor for assistance with cell sorting; and S. Shih and members of the lab for discussions and for critically reviewing the manuscript. VACD2 vector was provided by D. Kioussis (National Institute for Medical Research, London, UK), and wild-type and catalytically inactive Jak3 were from J. Ihle (St. Jude Children's Research Hospital). Supported by the Howard Hughes Medical Institute and the National Institutes of Health (AI-0466888 and AI-055502).
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Horng, T., Bezbradica, J. & Medzhitov, R. NKG2D signaling is coupled to the interleukin 15 receptor signaling pathway. Nat Immunol 8, 1345–1352 (2007). https://doi.org/10.1038/ni1524
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DOI: https://doi.org/10.1038/ni1524
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