Outcome and prognosis of kidney transplantation are determined by allograft rejection. Some evidences suggest an involvement of Th17 cells, a cell subset producing IL-17, in the pathogenesis of allograft rejection. In addition, an important role of regulatory T cells (Treg) suppressing acute rejection has been previously described. Although some experimental data directly link Th17 and/or Treg cells to rejection, most clinical data are limited to the detection of these cell subsets either by immunohistochemistry (IHC) or gene expression analysis of graft biopsy samples posing a significant problem for true cell counting, since IHC and gene expression analysis are limited in formal quantification of cell numbers.
Here, we established a fully quantitative DNA methylation-based method for Th17 cell counting. Treg cells were enumerated by analysing demethylation in FOXP3 locus as previously established by our group. Using this methodology, we analyzed renal allograft biopsy samples from patient with acute cellular rejection. Histological evaluation were performed according to Banff criteria. Biopsies of age, gender and immunosuppressive regimen matched kidney transplant patients were used as controls. Patients were clinically monitored with a median follow up of 8.6 years.
Our data showed a significantly higher number of Th17 cells in acute rejection biopsies (median 0.98 %; range 0.18%-1.67%) as compared to control biopsies (median 0.26%; range 0.06-0.47%) (p<0.05). In addition, we observed a significantly higher level of Treg cells infiltrating allografts with acute rejection (median 1.5%; range 0.5-3.7%) compared to control biopsies (median 0.1%; range 0.1-0.2%) (p<0.01).
In conclusion, we established a new method for analysis of Th17 cells, which allows characterization of cellular intragraft infiltrates in a specific, standardized, and a fully quantitative manner. Elevated number of Th17 cells found in kidney allograft biopsies with acute rejections confirms previous observations in animal models and provides new opportunities as a therapeutic target.
To cite this abstract in AMA style:Babel N, Baron U, Rosenberger C, Schindler R, Dziubianau M, Sefrin A, Olek S, Reinke P. Epigenetic Approach for Standardized and Quantitative Analysis of Renal Allograft Biopsies: Acute Cellular Rejection Is Characterized by Infiltration of Th17 and Treg Cells [abstract]. Am J Transplant. 2013; 13 (suppl 5). https://atcmeetingabstracts.com/abstract/epigenetic-approach-for-standardized-and-quantitative-analysis-of-renal-allograft-biopsies-acute-cellular-rejection-is-characterized-by-infiltration-of-th17-and-treg-cells/. Accessed October 27, 2020.
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