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Urine protein profiling identified alpha-1-microglobulin and haptoglobin as biomarkers for early diagnosis of acute allograft rejection following kidney transplantation

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Abstract

Purpose

Early diagnosis of acute rejection and effective immunosuppressive therapy lead to improvement in graft survival following kidney transplantation. In this study, we aimed to establish a urinary protein profile suitable to distinguish between patients with rejection and stable graft function and to predict acute rejection based on postoperatively collected urine samples. A further objective was to identify candidate proteins for the use as biomarkers in clinical practice.

Methods

Urine samples of 116 kidney recipients were included. Rejection was proven by biopsy (n = 58), and stable transplant function was monitored for at least 2 years (n = 58). Postoperative urine samples were collected between 3rd and 10th day following transplantation. Urinary protein profiles were obtained by surface-enhanced laser desorption/ionization time-of-flight mass spectrometry. Protein identification and validation were performed using multiplex fluorescence 2DE, peptide mass fingerprinting and enzyme-linked immunosorbent assay.

Results

A protein profile including four mass peaks differentiated acute rejection from stable transplants at the time point of rejection and at the postoperative state with 73 % sensitivity and 88 % specificity. Alpha-1-microglobulin (A1MG) and Haptoglobin (Hp) were identified as putative rejection biomarkers. Protein levels were significantly higher in postoperative urine from patients with rejection (A1MG 29.13 vs. 22.06 μg/ml, p = 0.001; Hp 628.34 vs. 248.57 ng/ml, p = 0.003). The combination of both proteins enabled the diagnosis of early rejection with 85 % sensitivity and 80 % specificity.

Conclusion

Protein profiling using mass spectrometry is suitable for noninvasive detection of rejection-specific changes following kidney transplantation. A specific protein profile enables the prediction of early acute allograft rejection in the immediate postoperative period. A1MG and Hp appear to be reliable rejection biomarkers.

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Acknowledgments

We like to thank Ms. Maria Pötsch for conducting MALDI-TOF-MS analysis and Mr. Saiful Miah for critical reading of this manuscript.

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical standard

The ethical approval was obtained from the Ethics Committee of the Medical Faculty of the University Jena. Samples were obtained with written consent.

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Authors and Affiliations

  1. Department of Urology, Jena University Hospital, Jena, Germany

    Beatrice Stubendorff, Stephanie Finke, Martina Walter, Torsten Gruschwitz, Thomas Steiner, Heiko Wunderlich & Kerstin Junker

  2. Clinic of Urology and Pediatric Urology, Saarland University Medical Center, Kirrberger Strasse, 66421, Homburg/Saar, Germany

    Beatrice Stubendorff & Kerstin Junker

  3. Department of Molecular and Applied Microbiology, Leibniz Institute for Natural Product Research and Infection Biology - Hans-Knöll-Institute and Integrated Research and Treatment Center – Center for Sepsis Control and Care (CSCC), Jena University Hospital, Friedrich Schiller University, Jena, Germany

    Olaf Kniemeyer

  4. Institute of Human Genetics, Core Unit Chip Application (CUCA), Jena University Hospital, Jena, Germany

    Ferdinand von Eggeling

  5. Department of Urology, Helios Hospital Erfurt, Erfurt, Germany

    Thomas Steiner

  6. KfH Kuratorium für Dialyse und Nierentransplantation e.V., KfH Nierenzentrum, Jena, Germany

    Undine Ott & Gunter Wolf

  7. Department of Internal Medicine III, Jena University Hospital, Jena, Germany

    Undine Ott & Gunter Wolf

  8. Clinic of Urology and Pediatric Urology, Eisenach St. Georg Hospital, Eisenach, Germany

    Heiko Wunderlich

Authors
  1. Beatrice Stubendorff
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  2. Stephanie Finke
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  3. Martina Walter
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  4. Olaf Kniemeyer
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  5. Ferdinand von Eggeling
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  6. Torsten Gruschwitz
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  7. Thomas Steiner
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  8. Undine Ott
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  9. Gunter Wolf
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  10. Heiko Wunderlich
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  11. Kerstin Junker
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Corresponding author

Correspondence to Kerstin Junker.

Additional information

Beatrice Stubendorff and Stephanie Finke have contributed equally to this work.

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Stubendorff, B., Finke, S., Walter, M. et al. Urine protein profiling identified alpha-1-microglobulin and haptoglobin as biomarkers for early diagnosis of acute allograft rejection following kidney transplantation. World J Urol 32, 1619–1624 (2014). https://doi.org/10.1007/s00345-014-1263-z

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  • DOI: https://doi.org/10.1007/s00345-014-1263-z

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