Abstract
The immune synapse is a complex cellular structure that enables cell–cell communications between immune cells, mainly at the interface of an effector T-cell and an antigen-presenting cell (APC) that expresses the appropriate peptide–MHC complexes. With progressive technological advances, there has been increasing interest in understanding molecular events that take place in motile T-lymphocytes at the immune synapse. Here, we provide an optimized method to induce the formation of an immune synapse between a T-cell and an APC in vitro. The experimental protocol described herein would be useful in characterizing the role of cell surface molecules and downstream signaling pathways in the context of cell-to-cell communications between T-cells and APCs.
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References
Dustin ML (2014) The immunological synapse. Cancer Immunol Res 2:1023–1033
Dustin ML, Bromley SK, Kan Z, Peterson DA, Unanue ER (1997) Antigen receptor engagement delivers a stop signal to migrating T lymphocytes. Proc Natl Acad Sci U S A 94:3909–3913
Huppa JB, Davis MM (2003) T-cell-antigen recognition and the immunological synapse. Nat Rev Immunol 3:973–983
Schubert DA, Gordo S, Sabatino JJ, Vardhana S, Gagnon E, Sethi DK, Seth NP, Choudhuri K, Reijonen H, Nepom GT, Evavold BD, Dustin ML, Wucherpfennig KW (2012) Self-reactive human CD4 T cell clones form unusual immunological synapses. J Exp Med 209:335–352
Zheng P, Bertolet G, Chen Y, Huang S, Liu D (2015) Super-resolution imaging of the natural killer cell immunological synapse on a glass-supported planar lipid bilayer. J Vis Exp (96):1–7
Purbhoo M, Liu H, Oddos S, Owen DM, Neil MA, Pageon SV, French PM, Rudd CE, Davis DM (2010) Dynamics of subsynaptic vesicles and surface microclusters at the immunological synapse. Sci Signal 3(121):ra36
Fischer AH, Jacobson KA, Rose J, Zeller R (2008) Preparation of slides and coverslips for microscopy. CSH Protoc 2008:pdb.prot4988
Acknowledgements
The authors acknowledge funding support from the Lee Kong Chian School of Medicine, Nanyang Technological University Singapore Start-Up Grant, the Ministry of Education (MOE) Singapore under its Singapore MOE Academic Research Fund (AcRF) Tier 2 Grant (MOE2017-T2-2-004) and the Industry Alignment Fund Pre-Positioning (IAF-PP) grant under the Wound Care Innovation for the Tropics IAF-PP Programme (H17/01/a0/0K9).
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Fazil, M.H.U.T., Kumar, P., Verma, N.K. (2019). A Protocol to Study T-Cell Signaling in an Immune Synapse by Microscopy. In: Verma, N. (eds) T-Cell Motility. Methods in Molecular Biology, vol 1930. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9036-8_15
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DOI: https://doi.org/10.1007/978-1-4939-9036-8_15
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-9035-1
Online ISBN: 978-1-4939-9036-8
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