Abstract
Extracellular signals are transmitted to intracellular targets thanks to a complex network of interacting proteins that regulate a large number of cellular processes. The signaling mechanisms that occur in CTLs are initiated by T cell receptors (TCRs) that recognize antigens presented in the groove of class I histocompatibility (MHC) molecules of target cells. The strength of this interaction and the consequent activation of intracellular mechanisms determine the fate of the developing T cells. The intracellular mediators of activation can be easily detected using a classical methodology such as western blot. Here we describe this technique bearing particular attention to these mediators using SNAP i.d., a newly developed protein detection system.
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References
Beal AM, Anikeeva N, Varma R, Cameron TO, Vasiliver-Shamis G, Norris PJ, Dustin ML, Sykulev Y (2009) Kinetics of early T cell receptor signaling regulate the pathway of lytic granule delivery to the secretory domain. Immunity 31:632–642
Robertson LK, Mireau LR, Ostergaard HL (2005) A role for phosphatidylinositol 3-kinase in TCR-stimulated ERK activation leading to paxillin phosphorylation and CTL degranulation. J Immunol 175:8138–8145
Pardo J, Buferne M, MartÃnez-Lorenzo MJ, Naval J, Schmitt-Verhulst AM, Boyer C, Anel A (2003) Differential implication of protein kinase C isoforms in cytotoxic T lymphocyte degranulation and TCR-induced Fas ligand expression. Int Immunol 15:1441–1450
Salmond RJ, Filby A, Qureshi I, Caserta S, Zamoyska R (2009) T-cell receptor proximal signaling via the Src-family kinases, Lck and Fyn, influences T-cell activation, differentiation, and tolerance. Immunol Rev 228:9–22
Welke J, Zavazava N (2002) P59(fyn) is upregulated in anergic CD8+ T cells. Hum Immunol 63:834–843
Jordan MS, Singer AL, Koretzky GA (2003) Adaptors as central mediators of signal transduction in immune cells. Nat Immunol 4:110–116
Fuller CL, Ravichandran KS, Braciale VL (1999) Phosphatidylinositol 3-kinase-dependent and -independent cytolytic effector functions. J Immunol 162:6337–6340
Falahati R, Leitenberg D et al (2007) Changes in the role of the CD45 protein tyrosine phosphatase in regulating Lck tyrosine phosphorylation during thymic development. J Immunol 178:2056–2064
Acknowledgments
This work was supported by grants from the MiUR (PON-REC ONEV 134/2011; FIRB RBAP11B2SX) and Regione Puglia (BISIMANE project, H31D08000030007).
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Cox, S.N. (2014). Intracellular Signaling of CTLs. In: Ranieri, E. (eds) Cytotoxic T-Cells. Methods in Molecular Biology, vol 1186. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1158-5_4
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DOI: https://doi.org/10.1007/978-1-4939-1158-5_4
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Publisher Name: Humana Press, New York, NY
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Online ISBN: 978-1-4939-1158-5
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