Reactivity of the immune system against donor antigens determines transplant-rejection after transplantation, and analyzing the alloimmune response has been shown to predict acute rejection in previous studies. The available methods for assessing alloreactivity are based on measurements of reactivity of recipient peripheral blood mononuclear cells (PBMCs) upon stimulation by stimulator cells derived from donor spleen cells or artificial third party stimulator cell banks. Despite its potential utility alloreactivity testing in clinical routine is restricted mostly by either low quantity (limited amount of donor spleen cells) or quality (lack of sufficient matching between HLA-bank and donor HLA) of stimulator cells. The aim of this study was to establish a renewable source of donor-derived cells for alloimmunity analysis in kidney transplantation.

Urinary samples were succecssfully used for generation of induced pluripotent stem cells in a previous study. Taking advantage of this, we established a protocol for the generation of patient-specific donor-derived stimulator cell lines using recipient's urine. The urinary samples were centrifugated, washed and the cell-pellet was seeded and cultivated with daily medium changes. The cultivated cells showed an epithelial phenotype and HLA genotyping revealed that >50% were donor-derived. Alloreactivity was proven by proliferation assay. For this, cell lines were incubated with the recipient's PBMCs in the ratios 1:1 and 1:5 (epithelial cells:PBMC) to test for the direct alloresponse. Lysed cell lines presentated on recipients PBMCs were used for indirect alloimmunity testing. Multi-color flow-cytometry measurement showed a dose-dependent CD3+ cell proliferation upon direct stimulation, and, at a lesser extent, after indirect stimulation.

In conclusion, we established a novel platform for monitoring allo-reactivity in kidney transplant patients. Further studies are required to assess clinical utility of this assay.

CITATION INFORMATION: Thieme C, Weist B, Schmück M, Reinke P, Babel N. Urine-Derived Cell Lines as a Novel Tool for Monitoring Allo-Reactivity in Kidney Transplant Patients. Am J Transplant. 2016;16 (suppl 3).

« Back to 2016 American Transplant Congress