Abstract: TH-PO0898
Immortalized Microvascular Renal Endothelial Cells as Target Cells for Antibody-Mediated Rejection Risk Assessment in Kidney Transplant Recipients
Session Information
- Transplantation: Basic Research
November 06, 2025 | Location: Exhibit Hall, Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Transplantation
- 2101 Transplantation: Basic
Authors
- Tiller, Gesa, University Medical Center Groningen, Department of Internal Medicine, Division of Nephrology, Groningen, Netherlands
- Altulea, Dania, University Medical Center Groningen, Department of Internal Medicine, Division of Nephrology, Groningen, Netherlands
- Lammerts, Rosa G.m., University Medical Center Groningen, Department of Laboratory Medicine, Transplantation Immunology, Groningen, Netherlands
- Dam, Wendy, University Medical Center Groningen, Department of Pathology and Medical Biology, Groningen, Netherlands
- Lais, Rahat, University Medical Center Groningen, Department of Internal Medicine, Division of Nephrology, Groningen, Netherlands
- Van Den Born, Jacob, University Medical Center Groningen, Department of Pathology and Medical Biology, Groningen, Netherlands
- Heeringa, Peter, University Medical Center Groningen, Department of Pathology and Medical Biology, Groningen, Netherlands
- Figueiredo, Constanca, Hannover Medical School,Institute of Transfusion Medicine and Transplant Engineering, Hannover, Germany
- Yard, Benito, University Medical Centre Mannheim, University of Heidelberg, 5 Fifth Department of Medicine (Nephrology/Endocrinology/Rheumatology/ Pneumology), Heidelberg, Germany
- Sanders, Jan-Stephan, University Medical Center Groningen, Department of Internal Medicine, Division of Nephrology, Groningen, Netherlands
- Berger, Stefan P., University Medical Center Groningen, Department of Internal Medicine, Division of Nephrology, Groningen, Netherlands
Background
Antibody-mediated rejection (ABMR) in kidney transplantation, driven by donor-specific antibodies (DSA), involves complement-dependent (CDC) and antibody-dependent cellular cytotoxicity (ADCC). Current models lack kidney-specific endothelial cell (EC) targets and donor diversity. This study developed immortalized microvascular renal ECs (iMPRECs) from donor perfusion fluid for use in flow cytometry-based crossmatch (FXM) and in vitro NK cell-mediated ADCC assays.
Methods
ECs were isolated via CD31 magnetic beads and immortalized using a lentiviral vector. Endothelial identity was confirmed through flow cytometry (expression of CD31, CD34, VEGFR-2, ET-1, and vWF), single-cell RNA sequencing, and in functional tube formation assay. iMPRECs were evaluated in flow crossmatch (FXM) and ADCC assays using sera with anti-HLA antibodies and complement for the CDC, and CD16+ oNK-1 cells for the ADCC. oNK-1 cells were kindly provided by Acepodia Inc. (California, USA).
Results
Seven iMPREC lines were generated and confirmed to have retained their endothelial markers and angiogenic capacity. FXM demonstrated significant IgM, IgG, and C3b binding post–anti-HLA exposure (P<0.05). ADCC assays showed elevated cytotoxicity against HLA class I–sensitized iMPRECs compared to controls (P=0.01), with a trend towards differences in relative cell cytotoxicity in the CDC and ADCC assay when comparing anti-HLA sera of different kidney transplant recipients.
Conclusion
We showed that iMPRECs provide organ-specific targets for assessing DSA-mediated cytotoxicity. FXM and ADCC assays capture complement- and NK cell–driven damage, revealing differential cytotoxic potential among anti-HLA sera. This model addresses limitations of non-renal ECs and supports personalized risk stratification. Future directions include expanding the cell bank and exploring HLA gene silencing to model DSA-negative rejection mechanisms.