One of the hallmarks of live cells is the asymmetric distribution of lipids across their plasma membrane. Changes in this asymmetry due to lipid “scrambling” result in phosphatidylserine exposure at the cell surface that is detected by annexin V staining. This alteration is observed during cell death processes such as apoptosis, and during physiological responses such as platelet degranulation and membrane repair. Previous studies have shown that activation of NK cells is accompanied by exposure of phosphatidylserine at the cell surface. While this response was thought to be indicative of ongoing NK cell death, it may also reflect the regulation of NK cell activation in the absence of cell death. Herein, we found that NK cell activation was accompanied by rapid phosphatidylserine exposure to an extent proportional to the degree of NK cell activation. Through enforced expression of a lipid scramblase, we provided evidence that activation-induced lipid scrambling in NK cells is reversible and does not lead to cell death. In contrast, lipid scrambling attenuates NK cell activation. This response was accompanied by reduced cell surface expression of activating receptors such as 2B4, and by loss of binding of Src family protein tyrosine kinases Fyn and Lck to the inner leaflet of the plasma membrane. Hence, lipid scrambling during NK cell activation is, at least in part, a physiological response that reduces the NK cell activation level. This effect is due to the ability of lipid scrambling to alter the distribution of membrane-associated receptors and kinases required for NK cell activation.
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Sunshine, H. & Iruela-Arispe, M. L. Membrane lipids and cell signaling. Curr. Opin. Lipidol. 28, 408–413 (2017).
van Meer, G., Voelker, D. R. & Feigenson, G. W. Membrane lipids: where they are and how they behave. Nat. Rev. Mol. Cell Biol. 9, 112–124 (2008).
Leventis, P. A. & Grinstein, S. The distribution and function of phosphatidylserine in cellular membranes. Annu. Rev. Biophys. 39, 407–427 (2010).
Bevers, E. M. & Williamson, P. L. Getting to the outer leaflet: physiology of phosphatidylserine exposure at the plasma membrane. Physiol. Rev. 96, 605–645 (2016).
Balasubramanian, K. & Schroit, A. J. Aminophospholipid asymmetry: a matter of life and death. Annu. Rev. Physiol. 65, 701–734 (2003).
Gong, Y. N. et al. ESCRT-III acts downstream of MLKL to regulate necroptotic cell death and its consequences. Cell 169, 286–300 e216 (2017).
Neumann, B. et al. EFF-1-mediated regenerative axonal fusion requires components of the apoptotic pathway. Nature 517, 219–222 (2015).
Dillon, S. R., Mancini, M., Rosen, A. & Schlissel, M. S. Annexin V binds to viable B cells and colocalizes with a marker of lipid rafts upon B cell receptor activation. J. Immunol. 164, 1322–1332 (2000).
Elliott, J. I. et al. Membrane phosphatidylserine distribution as a non-apoptotic signalling mechanism in lymphocytes. Nat. Cell Biol. 7, 808–816 (2005).
Fischer, K. et al. Antigen recognition induces phosphatidylserine exposure on the cell surface of human CD8+ T cells. Blood 108, 4094–4101 (2006).
Ma, Y., Poole, K., Goyette, J. & Gaus, K. Introducing membrane charge and membrane potential to T cell signaling. Front Immunol. 8, 1513 (2017).
Rival, C. M. et al. Phosphatidylserine on viable sperm and phagocytic machinery in oocytes regulate mammalian fertilization. Nat. Commun. 10, 4456 (2019).
Nagata, S. & Segawa, K. Sensing and clearance of apoptotic cells. Curr. Opin. Immunol. 68, 1–8 (2020).
Segawa, K. & Nagata, S. An apoptotic ‘Eat Me’ signal: phosphatidylserine exposure. Trends Cell Biol. 25, 639–650 (2015).
Ruhl, S. et al. ESCRT-dependent membrane repair negatively regulates pyroptosis downstream of GSDMD activation. Science 362, 956–960 (2018).
Nagata, S. Apoptosis and clearance of apoptotic cells. Annu. Rev. Immunol. 36, 489–517 (2018).
Fujii, T., Sakata, A., Nishimura, S., Eto, K. & Nagata, S. TMEM16F is required for phosphatidylserine exposure and microparticle release in activated mouse platelets. Proc. Natl Acad. Sci. USA 112, 12800–12805 (2015).
Wu, N. et al. Critical role of lipid scramblase TMEM16F in phosphatidylserine exposure and repair of plasma membrane after pore formation. Cell Rep. 30, 1129–1140 e1125 (2020).
Yeung, T. et al. Receptor activation alters inner surface potential during phagocytosis. Science 313, 347–351 (2006).
O’Donnell, V. B., Rossjohn, J. & Wakelam, M. J. Phospholipid signaling in innate immune cells. J. Clin. Investig. 128, 2670–2679 (2018).
Resh, M. D. Myristylation and palmitylation of Src family members: the fats of the matter. Cell 76, 411–413 (1994).
Resh, M. D. Targeting protein lipidation in disease. Trends Mol. Med. 18, 206–214 (2012).
Yeung, T. et al. Membrane phosphatidylserine regulates surface charge and protein localization. Science 319, 210–213 (2008).
Segawa, K. et al. Caspase-mediated cleavage of phospholipid flippase for apoptotic phosphatidylserine exposure. Science 344, 1164–1168 (2014).
Suzuki, J., Imanishi, E. & Nagata, S. Exposure of phosphatidylserine by Xk-related protein family members during apoptosis. J. Biol. Chem. 289, 30257–30267 (2014).
Suzuki, J., Umeda, M., Sims, P. J. & Nagata, S. Calcium-dependent phospholipid scrambling by TMEM16F. Nature 468, 834–838 (2010).
Vivier, E. et al. Innate or adaptive immunity? The example of natural killer cells. Science 331, 44–49 (2011).
Guo, H. et al. Deletion of Slam locus in mice reveals inhibitory role of SLAM family in NK cell responses regulated by cytokines and LFA-1. J. Exp. Med. 213, 2187–2207 (2016).
Zhang, Z. et al. DNAM-1 controls NK cell activation via an ITT-like motif. J. Exp. Med. 212, 2165–2182 (2015).
Cruz-Munoz, M. E., Dong, Z., Shi, X., Zhang, S. & Veillette, A. Influence of CRACC, a SLAM family receptor coupled to the adaptor EAT-2, on natural killer cell function. Nat. Immunol. 10, 297–305 (2009).
Sandilands, E., Brunton, V. G. & Frame, M. C. The membrane targeting and spatial activation of Src, Yes and Fyn is influenced by palmitoylation and distinct RhoB/RhoD endosome requirements. J. Cell Sci. 120, 2555–2564 (2007).
Stacey, M. A., Marsden, M., Wang, E. C., Wilkinson, G. W. & Humphreys, I. R. IL-10 restricts activation-induced death of NK cells during acute murine cytomegalovirus infection. J. Immunol. 187, 2944–2952 (2011).
Nakamura, K. et al. Fratricide of natural killer cells dressed with tumor-derived NKG2D ligand. Proc. Natl Acad. Sci. USA 110, 9421–9426 (2013).
Rudd-Schmidt, J. A. et al. Lipid order and charge protect killer T cells from accidental death. Nat. Commun. 10, 5396 (2019).
Zech, T. et al. Accumulation of raft lipids in T-cell plasma membrane domains engaged in TCR signalling. EMBO J. 28, 466–476 (2009).
Petkovic, M., Oses-Prieto, J., Burlingame, A., Jan, L. Y. & Jan, Y. N. TMEM16K is an interorganelle regulator of endosomal sorting. Nat. Commun. 11, 3298 (2020).
Hu, Y. et al. Scramblase TMEM16F terminates T cell receptor signaling to restrict T cell exhaustion. J. Exp. Med. 213, 2759–2772 (2016).
Wu, N. et al. A hematopoietic cell-driven mechanism involving SLAMF6 receptor, SAP adaptors and SHP-1 phosphatase regulates NK cell education. Nat. Immunol. 17, 387–396 (2016).
Lemay, S., Davidson, D., Latour, S. & Veillette, A. Dok-3, a novel adapter molecule involved in the negative regulation of immunoreceptor signaling. Mol. Cell Biol. 20, 2743–2754 (2000).
Veillette, A., Thibaudeau, E. & Latour, S. High expression of inhibitory receptor SHPS-1 and its association with protein-tyrosine phosphatase SHP-1 in macrophages. J. Biol. Chem. 273, 22719–22728 (1998).
This work was supported by grants from the Canadian Institutes of Health Research (CIHR) MT-14429, MOP-82906, and FDN-143338 to A.V., and NSFC-31870863 from the National Natural Science Foundation of China to N.W. N.W. was supported by a Postdoctoral Fellowship from Fonds de recherche du Québec Santé (FRQS). A.V. is the Canada Research Chair on Signaling in the Immune System.
A.V. received a contract from Bristol Myers-Squibb to study the mechanism of action of the anti-SLAMF7 monoclonal antibody elotuzumab in multiple myeloma. He was also a consultant for Boehringer-Ingelheim on the topic of the SIRPα-CD47 blockade in anticancer immunotherapy. The authors declare no competing interests.
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Wu, N., Song, H. & Veillette, A. Plasma membrane lipid scrambling causing phosphatidylserine exposure negatively regulates NK cell activation. Cell Mol Immunol 18, 686–697 (2021). https://doi.org/10.1038/s41423-020-00600-9
- NK cell activation
- Phosphatidylserine exposure
- Lipid scrambling