Abstract
During the course of an immune response in kidney transplantation, distinct functional subsets of effector and regulatory T cells are generated in the lymphoid compartment following the differentiation of T cells under the influence of specific cytokines. In addition to effector cells inflammation in a CXCR3-independent manner, a CXCR3-chemokine dependent pathway exacerbates these inflammatory processes in this scenario. Indeed, the upregulation of CXCR3 ligands mediates the mobilization of effector CTLs to the peripheral site of infection. The immediate upregulation of CXCR3 on CD4+ CTLs and CD8+ CTL following DC activation makes this an interesting proposal, as activated T cells are recruited into lymphoid and noncanonical lymphoid compartments. The importance of tissue in vivo characterization is emerged as a central topic in kidney transplantation.
In the following method we describe a protocol based on immunohistochemistry (IHC) and immunofluorescence/confocal microscopy along with strategies of overall signals quantification and positive cells quantification (Aperio, Adobe Photoshop). Through an in vivo approach, we focus on those changes that result relevant during immune response in transplantation. Although less quantitative than other methods, the information gained from IHC combined with microscopy provides a “picture” that can help to address our subsequent experiments. The advantage of this protocol consists in the possibility to evidence CTLs tissue accumulation and to investigate the different areas (tubular, glomerular, and interstitial) of the graft directly affected by CTLs-specific activity.
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References
Strehlau J, Pavlakis M, Lipman M et al (1997) Quantitative detection of immune activation transcripts as a diagnostic tool in kidney transplantation. Proc Natl Acad Sci U S A 94:695–700
Lipman ML, Stevens AC, Strom TB (1994) Heightened intragraft CTL gene expression in acutely rejecting renal allografts. J Immunol 152:5120–5127
Kummer JA, Wever PC, Kamp AM (1995) Expression of granzyme A and B proteins by cytotoxic lymphocytes involved in acute renal allograft rejection. Kidney Int 47:70–77
Lipman ML1, Shen Y, Jeffery JR, Gough J, McKenna RM, Grimm PC, Rush DN (1998) Immune-activation gene expression in clinically stable renal allograft biopsies: molecular evidence for subclinical rejection. Transplantation 27;66(12):1673–1681
Hoffmann SC, Hale DA, Kleiner DE, Mannon RB, Kampen RL, Jacobson LM, Cendales LC, Swanson SJ, Becker BN, Kirk AD (2005) Functionally significant renal allograft rejection is defined by transcriptional criteria. Am J Transplant 5:573–581
Bromley SK, Mempel TR, Luster AD (2008) Orchestrating the orchestrators: chemokines in control of T cell traffic. Nat Immunol 9:970–980
Campanella GS, Grimm J, Manice LA et al (2006) Oligomerization of CXCL10 is necessary for endothelial cell presentation and in vivo activity. J Immunol 177:6991–6998
Qin S, Rottman JB, Myers P et al (1998) The chemokine receptors CXCR3 and CCR5 mark subsets of T cells associated with certain inflammatory reactions. J Clin Invest 101:746–754
Menke J, Zeller GC, Kikawada E et al (2008) CXCL9, but not CXCL10, promotes CXCR3-dependent immune-mediated kidney disease. J Am Soc Nephrol 19:1177–1189
Steinmetz OM, Turner JE, Paust HJ et al (2009) CXCR3 mediates renal Th1 and Th17 immune response in murine lupus nephritis. J Immunol 183:4693–4704
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Rascio, F., Divella, C., Grandaliano, G. (2014). CTL and Transplantation: Tissue In Vivo Characterization. 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_16
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DOI: https://doi.org/10.1007/978-1-4939-1158-5_16
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-1157-8
Online ISBN: 978-1-4939-1158-5
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