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Proteinuria after renal transplantation: Diagnosis with highly sensitive silver stain in sodium dodecylsulphate-polyacrylamide gradient gel electrophoresis

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Summary

Sodium dodecylsulphate-polyacrylamide gel electrophoresis (SDS-PAGE) of urinary proteins is becoming increasingly significant in monitoring renal allograft recipients. After conventional Coomassie Blue staining, changes in renal proteinuric patterns during acute renal transplant rejection could not be seen in many cases. Discrimination between rejection and nephrotoxic side effects of cyclosporine overdosage was also not possible. We therefore examined the possible advantages of a highly sensitive silver staining technique in this study. A total of 734 urine samples obtained from 38 patients after allogenic kidney transplantation were examined by SDS-PAGE with consecutive Coomassie Blue and silver staining. Twenty-two histologically proven rejections and 20 cyclosporine overdosage episodes were diagnosed in these patients within a time period of up to 524 days after transplantation. No changes in proteinuric patterns were seen in 9 of 22 patients after Coomassie stain during rejection, and only 12 cases showed a rise of glomerular protein bands, whereas silver stain revealed an increase in 19 of 22 cases. Discrimination between cyclosporine overdosage and rejection was possible with a probability ofp<0.001 after silver stain when using the changes in the number of glomerular protein bands as a criterion. These findings suggest that application of a highly sensitive silver stain instead of the conventional Coomassie stain after SDS-PAGE reflects considerable progress in monitoring renal allograft recipients.

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Abbreviations

SDS-PAGE:

Sodium dodecylsulphate-polyacrylamide gel electrophoresis

References

  1. 1.

    Andrassy K (1977) Diagnostik der akuten Abstoßung mit Hilfe von Laboruntersuchungen unter besonderer Berück-sichtigung der Polyacrylamidgel-Elektrophorese. In: Lenhard V, Dreikorn K, Ritz E (Hrsg) Aktuelle prä- und postoperative Probleme der Nierentransplantation. Hygieneplan, Frankfurt, S 105–111

  2. 2.

    Boesken WH (1985) Bedeutung der Proteinurie — Analyse für die nephrologische Diagnostik. Mitt Klin Nephrol 14:96–111

  3. 3.

    Burghard R, Galaske RG, Offner G, Ehrich JHH, Leititis JU, Brodehl J (1986) Urinary protein analysis in the early detection of acute rejection episodes after renal transplantation in children. Clin Nephrol 26:1–6

  4. 4.

    Frey BM, Frey FJ, Wegmüller E, Kodier G (1977) Proteinurie nach Nierentransplantation. Schweiz Med Wochenschr 107:1181–1190

  5. 5.

    Heukeshoven J, Dernick R (1986) Neue Ergebnisse zum Mechanismus der Silberfärbung. Elektrophorese Forum 86:22–27

  6. 6.

    Kirchherr H, Schiwara HW (1985) Quantitative Proteinbestimmung im Harn mit einer empfindlichen Streulichtmethode. J Clin Chem Clin Biochem 23:57–62

  7. 7.

    Krishna KS, Pandey AP, Kirubakaran MG, Kanagasabapathy AS (1987) Urinary protein/creatinine ratio as an inidcator of allograft function following live related donor renal transplantation. Clin Chim Acta 163:51–61

  8. 8.

    Lapin A, Zyman H, Gabl F (1986) Monitoring of urinary proteins by SDS-Electrophoresis in kidney transplant patients. J Clin Chem Clin Biochem 24:221–225

  9. 9.

    Lillehoj EP, Poulik MD (1986) Normal and abnormal aspects of proteinuria. Exp Pathol 29:1–28

  10. 10.

    Lison AE, Brandt U, Weisshaar CH, Meyer E, Buchholz B, Zastrow F (1985) Frühzeitige Diagnose der Abstoßungsreaktion nach Nierentransplantation mit Hilfe der SDS-Disk-Elektrophorese? Nieren- und Hochdruckkrankheiten 14:537–542

  11. 11.

    Lison AE, Rolf N, Igel G, Koch J, Meyer E (1986) Nephrotoxische Effekte unter Langzeittherapie mit Cyclosporin A. Nieren- und Hochdruckkrankheiten 15:417

  12. 12.

    Lison AE, Doedt E, Meyer E (1989) Automatic microdisc electrophoresis for urinary protein analysis: a comparison with standard analysis. Clin Nephrol 32:40–45

  13. 13.

    Odlind B, Backman U, Forbes MA, Cooper EH (1988) Proteinuria after renal transplantation. Contrib Nephrol 68:149–156

  14. 14.

    Reiber H (1983) Kinetics of protein agglomeration. A nephelometric method for the determination of total protein in biological samples. J Biochem Biophys Methods 7:153–160

  15. 15.

    Schiwara HW, Hebell T, Kirchherr H, Postel W, Weser J, Görg A (1986) Ultrathin-layer sodium dodecyl sulfatepolyacrylamide gradient gel electrophoresis and silver staining of urinary proteins. Electrophoresis 7:496–505

  16. 16.

    Sethi K, First MR, Pesce AJ, Fidler JP, Pollak VE (1977) Proteinuria following renal transplantation. Nephron 18:49–59

  17. 17.

    Steinhoff J, Feddersen A, Wood WG, Hoyer J, Sack K (1991) Glomerular proteinuria as an early sign of renal transplant rejection. Clin Nephrol 35:255–262

  18. 18.

    Vathsala A, Verani R, Schoenberg L, Lewis RM, Van Buren CT, Kerman RH, Kahan BD (1990) Proteinuria in cyclosporine-treated renal transplant recipients. Transplantation 49:35–41

  19. 19.

    Weber MH (1988) Urinary protein analysis. J Chromatogr 429:315–344

  20. 20.

    Woo J, Floyd M, Cannon DC (1981) Albumin andβ2- microglobulin radioimmunoassay applied to monitoring of renal allograft function and in differentiating glomerular and tubular diseases. Clin Chem 27:709–713

  21. 21.

    Wood WG, Herhahn D, Steinhoff J, Feddersen A, Schulz E, Sack K (1990) The diagnostic relevance of specific urinary proteins after renal transplantation. Ärztl Lab 36:260–267

  22. 22.

    Wurster U, Ehrich JHH (1985) Silberfärbung von Urinproteinen. Nieren- und Hochdruckkrankheiten 14:491–496

  23. 23.

    Zastrow F, Buchholz B, Lison AE, Gottschalk IS (1981) Die Abstoßungsreaktion der transplantierten Niere — ihre Frühdiagnose und Behandlung. Med Welt 47:1787–1789

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Kirschstein, M., Jensen, R., Schröder, R. et al. Proteinuria after renal transplantation: Diagnosis with highly sensitive silver stain in sodium dodecylsulphate-polyacrylamide gradient gel electrophoresis. Klin Wochenschr 69, 847–852 (1991). https://doi.org/10.1007/BF01649458

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Key words

  • Renal transplantation
  • Electrophoresis
  • Silver stain
  • Coomassie stain
  • Rejection
  • Proteinuria
  • Cyclosporine