Abstract
Recent advances in imaging technology have enabled significant advances in the study of NK cell cytotoxic effector function through quantitative analysis of the NK cell immunological synapse. This can include the use of high- and super-resolution microscopy to quantify dynamics of cytoskeletal elements and the role they play in the regulation and execution of NK cell directed secretion. Here we describe a protocol for the recapitulation of the NK cell lytic synapse on glass, the acquisition of microscopy images, and suggested approaches for the processing and analysis of microscopy data.
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References
Mace EM, Dongre P, Hsu HT et al (2014) Cell biological steps and checkpoints in accessing NK cell cytotoxicity. Immunol Cell Biol 92:245–255
Lagrue K, Carisey A, Oszmiana A et al (2013) The central role of the cytoskeleton in mechanisms and functions of the NK cell immune synapse. Immunol Rev 256:203–221
Mace EM, Orange JS (2012) New views of the human NK cell immunological synapse: recent advances enabled by super- and high-resolution imaging techniques. Front Immunol 3:421
Brown AC, Oddos S, Dobbie IM et al (2011) Remodelling of cortical actin where lytic granules dock at natural killer cell immune synapses revealed by super-resolution microscopy. PLoS Biol 9, e1001152
Rak GD, Mace EM, Banerjee PP et al (2011) Natural killer cell lytic granule secretion occurs through a pervasive actin network at the immune synapse. PLoS Biol 9, e1001151
Gong JH, Maki G, Klingemann HG (1994) Characterization of a human cell line (NK-92) with phenotypical and functional characteristics of activated natural killer cells. Leukemia 8:652–658
Comrie WA, Babich A, Burkhardt JK (2015) F-actin flow drives affinity maturation and spatial organization of LFA-1 at the immunological synapse. J Cell Biol 208:475–491
Pageon SV, Cordoba SP, Owen DM et al (2013) Superresolution microscopy reveals nanometer-scale reorganization of inhibitory natural killer cell receptors upon activation of NKG2D. Sci Signal 6:ra62
Mace EM, Wu WW, Ho T et al (2012) NK cell lytic granules are highly motile at the immunological synapse and require F-actin for post-degranulation persistence. J Immunol 189:4870–4880
Mace EM, Orange JS (2014) Lytic immune synapse function requires filamentous actin deconstruction by Coronin 1A. Proc Natl Acad Sci U S A 111:6708–6713
Toomre D, Bewersdorf J (2010) A new wave of cellular imaging. Annu Rev Cell Dev Biol 26:285–314
Mace EM, Orange JS (2014) Visualization of the immunological synapse by dual color time-gated stimulated emission depletion (STED) nanoscopy. J Vis Exp
Orange JS, Roy-Ghanta S, Mace EM et al (2011) IL-2 induces a WAVE2-dependent pathway for actin reorganization that enables WASp-independent human NK cell function. J Clin Invest 121:1535–1548
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Mace, E.M., Orange, J.S. (2016). High- and Super-Resolution Microscopy Imaging of the NK Cell Immunological Synapse. In: Somanchi, S. (eds) Natural Killer Cells. Methods in Molecular Biology, vol 1441. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3684-7_12
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DOI: https://doi.org/10.1007/978-1-4939-3684-7_12
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-3682-3
Online ISBN: 978-1-4939-3684-7
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