Donor-Derived Cell-Free DNA for Diagnosing Subclinical Rejection: A Pilot Multicenter Study in Japanese Living-Donor Kidney Transplantation

Clin Transplant. 2025 Jul;39(7):e70221. doi: 10.1111/ctr.70221.

Abstract

Background: Donor-derived cell-free DNA (dd-cfDNA) offers a non-invasive biomarker for detecting allograft injury. However, its performance in predominantly living-donor transplant settings in Asian populations remains underexplored.

Methods: In a multicenter prospective pilot study, 73 living-donor kidney transplant (LKT) recipients from Japan underwent plasma dd-cfDNA testing and protocol biopsies approximately 1 year post-transplant. Diagnostic performance of dd-cfDNA for subclinical antibody-mediated rejection (AMR) was evaluated at 1.0% and 0.5% cut-off thresholds.

Results: Eight patients (11.0%) were diagnosed with subclinical active AMR. Patients with dd-cfDNA ≥1.0% showed significantly higher incidence of AMR compared to those with dd-cfDNA <1.0% (80% vs. 5.9%, p < 0.001). ROC analysis revealed an AUC of 0.85 for dd-cfDNA in diagnosing AMR. Lowering the threshold to 0.5% improved sensitivity while maintaining high specificity. Patients with persistent donor-specific antibodies (DSA) at 1-year follow-up exhibited significantly higher dd-cfDNA levels. Neither HLA nor ABO incompatibility nor donor-recipient relation significantly affected dd-cfDNA levels.

Conclusion: dd-cfDNA demonstrates high negative predictive value and complements DSA testing for early detection of subclinical AMR in living-donor transplants. This pilot study provides foundational evidence for dd-cfDNA-based surveillance strategies in Asian cohorts.

Keywords: biomarker; donor‐derived cell‐free DNA; donor‐specific antibody; subclinical antibody‐mediated rejection; surveillance biopsy.

Publication types

  • Multicenter Study

MeSH terms

  • Adult
  • Aged
  • Biomarkers* / blood
  • Cell-Free Nucleic Acids* / blood
  • Cell-Free Nucleic Acids* / genetics
  • East Asian People
  • Female
  • Follow-Up Studies
  • Glomerular Filtration Rate
  • Graft Rejection* / blood
  • Graft Rejection* / diagnosis
  • Graft Rejection* / etiology
  • Graft Survival
  • Humans
  • Isoantibodies / blood
  • Japan
  • Kidney Failure, Chronic* / surgery
  • Kidney Transplantation* / adverse effects
  • Living Donors*
  • Male
  • Middle Aged
  • Pilot Projects
  • Prognosis
  • Prospective Studies
  • Risk Factors

Substances

  • Cell-Free Nucleic Acids
  • Biomarkers
  • Isoantibodies

Supplementary concepts

  • Japanese people