Abstract
Cytokines of the TNF superfamily, particularly TNF itself, are important mediators of inflammatory leukocyte recruitment and activation in parenchymal organs. In inflammatory kidney diseases, leukocytes accumulate in glomeruli and the tubulointerstitium, leading to glomerulonephritis and tubulointerstitial nephritis, respectively. In particular, glomeruli can be the target of organ-threatening leukocyte-mediated inflammation. As microvasculatures of the glomerulus and the tubulointerstitium differ markedly in their structural and functional properties, recruitment and subsequent activation of leukocytes to these sites occur via distinct mechanisms. To understand the pathways and mediators of leukocyte-driven inflammation in the kidney it is therefore essential to analyze glomerular and tubulointerstitial leukocyte recruitment in a compartment-specific way. The protocol presented here describes an easy and rapid technique that allows compartment-specific quantitation and qualitative analysis of leukocytes present in glomeruli and tubulointerstitial tissue by flow cytometry after separation of these tissue compartments.
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Acknowledgements
The author thanks Andreas Blutke, Institute for Veterinary Pathology, Ludwig-Maximilians-University Munich for sharing his experience with the magnetic glomerular isolation procedure, Werner Vielhauer for designing the pressure-controlled perfusion device, and Nuru Eltrich for his continued technical support. Sibylle von Yorck is acknowledged for providing the excellent graphical illustration of the glomerular isolation procedure. This work was supported by DFG grant VI 231/2-1 to V. V.
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Vielhauer, V. (2014). Compartment-Specific Flow Cytometry for the Analysis of TNF-Mediated Recruitment and Activation of Glomerular Leukocytes in Murine Kidneys. In: Bayry, J. (eds) The TNF Superfamily. Methods in Molecular Biology, vol 1155. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0669-7_15
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DOI: https://doi.org/10.1007/978-1-4939-0669-7_15
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-0668-0
Online ISBN: 978-1-4939-0669-7
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