Fas ligand (FasL), an apoptosis-inducing member of the TNF cytokine family, and its receptor Fas are critical for the shutdown of chronic immune responses1,2,3 and prevention of autoimmunity4,5. Accordingly, mutations in their genes cause severe lymphadenopathy and autoimmune disease in mice6,7 and humans8,9. FasL function is regulated by deposition in the plasma membrane and metalloprotease-mediated shedding10,11. Here we generated gene-targeted mice that selectively lack either secreted FasL (sFasL) or membrane-bound FasL (mFasL) to resolve which of these forms is required for cell killing and to explore their hypothesized non-apoptotic activities. Mice lacking sFasL (FasLΔs/Δs ) appeared normal and their T cells readily killed target cells, whereas T cells lacking mFasL (FasLΔm/Δm ) could not kill cells through Fas activation. FasLΔm/Δm mice developed lymphadenopathy and hyper-gammaglobulinaemia, similar to FasLgld/gld mice, which express a mutant form of FasL that cannot bind Fas, but surprisingly, FasLΔm/Δm mice (on a C57BL/6 background) succumbed to systemic lupus erythematosus (SLE)-like autoimmune kidney destruction and histiocytic sarcoma, diseases that occur only rarely and much later in FasLgld/gld mice. These results demonstrate that mFasL is essential for cytotoxic activity and constitutes the guardian against lymphadenopathy, autoimmunity and cancer, whereas excess sFasL appears to promote autoimmunity and tumorigenesis through non-apoptotic activities.
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We thank Genentech, in particular A. Ashkenazi and S. Masters, for Fas-Fc fusion proteins. We also thank our colleagues at the Walter and Eliza Hall Institute of Medical Research (Melbourne), G. Siciliano, N. Iannarella, J. Coughlin for animal care; J. Corbin for automated blood analysis; B. Helbert and C. Young for genotyping; S. Mihajlovic, E. Tsui, A. Hasanein, V. Babo and K. Weston for histological sections; A. Light and K. O’Donnell for help with antibody measurements; and S. Drake for help with cytokine quantification. We also thank J. Melny (Royal Melbourne Hospital) for measuring anti-DNA antibody levels; A. Banerjee, S. Gerondakis and R. Gugasyan (Burnet Medical Research Institute, Prahran) for antibodies and advice; J. Silke and L. Wong (La Trobe University, Bundoora) for TNFα; P. Morgan for assistance with protein purification; and A. Silva and J. Sharkey for help with animal procedures (Peter MacCallum Cancer Centre). This work was supported in part by the NHMRC (programme numbers 461221 and 454569, a CJ Martin fellowship to N.M.H. and an RD Wright fellowship to L.A.O’R.). We also acknowledge support from the IRIISS (grant numbers 361646 and 257502 to P.B.), the Victorian State Government (an OIS grant), the Leukemia and Lymphoma Society (SCOR grant number 7015), the NIH (grant numbers CA043540-18 and CA80188-6), the JDRF/NHMRC, the Association for International Cancer Research, the Charles and Sylvia Viertel Charitable Foundation (to P.B.) and the Leukemia Research Foundation.
Author Contributions L.A.O’R. planned and performed most experiments and wrote the manuscript. L.T., L.L., E.A.K., S.G., W.D.F., N.M.H., D.M.T., J.-G.Z., G.T.B., M.J.S., P.B. and L.R. contributed to the planning and execution of the experiments and the writing of the manuscript. A.S. conceived the study, planned experiments and wrote the manuscript.
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O’ Reilly, L., Tai, L., Lee, L. et al. Membrane-bound Fas ligand only is essential for Fas-induced apoptosis. Nature 461, 659–663 (2009). https://doi.org/10.1038/nature08402
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