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. 2019 Dec 13;9:19037. doi: 10.1038/s41598-019-55442-x

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Assessment of alloreactive T cells using urine-derived donor TEC-stimulation shows higher sensitivity compared to donor splenocytes-stimulation. (a) Splenocytes express higher levels of HLA-DR than TEC. Representative plots demonstrating flow cytometric analysis of HLA-ABC and HLA-DR expression on untreated and 24 h IFNγ and TNFα treated urine-derived TEC or splenocytes. Both cell types were derived from the same deceased donor. (b,c) Frequencies of HLA-ABC (b) and HLA-DR (c) expressing living splenocytes and treated living TEC determined by flow cytometry (n = 4). (d) TEC elicit a higher alloreactivity than splenocytes from the same donor. PBMCs of four kidney-allograft recipients of different time points (n = 15) were stimulated with IFNγ and TNFα treated TEC and lysed TEC or splenocytes and lysed splenocytes, both derived from the kidney-transplant donor. Specific activation was assessed by flow cytometric measurements of the activation marker CD154 and CD137 expression on CD4⁺ T cells. Statistical analysis was done with Wilcoxon matched-pairs signed rank test.