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In Vitro Pentamer Formation as a Biomarker of Tacrolimus-Related Immunosuppressive Activity after Liver Transplantation

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Published/Copyright: November 12, 2000
Clinical Chemistry and Laboratory Medicine (CCLM)
From the journal Volume 38 Issue 11

Abstract

Therapeutic drug monitoring of tacrolimus (FK) is widely performed to assist adjustments of drug dosage but may be an inadequate surrogate of the immunosuppression induced. The aim of this investigation was to develop an alternative method for measuring FK-related immunosuppressive activity in blood samples from liver transplant recipients. A pentamer formation assay (PFA) was devised based on the attachment of the 12 kDa FK-binding protein (FKBP12) to microtitre plates in the presence of calcineurin, calmodulin, Ca++ and FK. Pentamer formation could be detected at FK concentrations ≥0.2 μg/l by optimising assay conditions, particularly by including Ca++ (0.5 mM) only during the formation of the pentameric complex. Three methods (blood lysis, proteolytic digestion and use of commercial solutions used in a microparticle enzyme immunoassay (MEIA) technique) were incompatible with PFA measurements after extracting immunosuppressive FK-related material from patients' blood samples. However, therapeutic amounts of FK-related material could be quantified by the PFA assay after extraction of blood samples with methanol. There was a moderate correlation (r = 0.689) of FK equivalents assayed by PFA with results using MEIA in 56 blood samples from 14 liver graft recipients, but no obvious relationship of results to variables reflecting their clinical status.

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Published Online: 2000-11-12
Published in Print: 2000-11-12

Copyright © 2000 by Walter de Gruyter GmbH & Co. KG

Articles in the same Issue

  1. European Meeting on Biomarkers of Organ Damage and Dysfunction Selected Papers from the Meeting Held in Cambridge, UK April 3rd-7th, 2000
  2. Kidney Retrieval Conditions Influence Damage to Renal Medulla: Evaluation by Proton Nuclear Magnetic Resonance (NMR) Spectroscopy
  3. Citrate, Acetate and Renal Medullary Osmolyte Excretion in Urine as Predictor of Renal Changes after Cold Ischaemia and Transplantation
  4. Comparison of Proteolytic Enzymes and Glutathione S-Transferase Levels in Non-Heart-Beating Donors' (NHBD) Kidney Perfusates
  5. Assessment of Non-Heart-Beating Donor (NHBD) Kidneys for Viability on Machine Perfusion
  6. Low Levels of Urinary Albumin Excretion Are Associated with Cardiovascular Risk Factors in the General Population
  7. Evaluation of a Fully Automated Assay to Measure C-Telopeptide of Type I Collagen in Serum
  8. Bone Turnover Markers and Estradiol Level in Postmenopausal Women
  9. External Quality Assessment of Bone Metabolism Marker Assays. Initial Experiences in a UK NEQAS Programme
  10. The Monoethylglycinexylidide (MEGX) Test as a Marker of Hepatic Dysfunction in Septic Patients with Pneumonia
  11. Transforming Growth Factor-β1 as a Surrogate Marker of Hepatic Dysfunction in Chronic Liver Diseases
  12. A New Approach to the Evaluation of Liver Graft Function by Nuclear Magnetic Resonance Spectroscopy. A Comparative Study between Euro-Collins and University of Wisconsin Solutions
  13. Metallothionein and Heat Shock Protein Expression during Acute Liver Injury and Regeneration in Rats
  14. Biochemical Evaluation of Patients with Acute Pancreatitis
  15. Screening for Acute Myocardial Injury: Creatine Kinase Is Comparable to Myoglobin
  16. Different Time Frames for the Occurrence of Elevated Levels of Cardiac Troponin T and C-Reactive Protein in Patients with Acute Myocardial Infarction
  17. Determination of Decision Limits for ACS:Systems Cardiac Troponin I
  18. Stability of Cardiac Troponins and CK-MB Mass Isoenzyme
  19. Cytokines as Plasma Markers of Abdominal Aortic Aneurysm
  20. The Early Fall in Levels of S-100 β in Traumatic Brain Injury
  21. Protein S-100β in Brain and Serum after Deep Hypothermic Circulatory Arrest in Rabbits: Relationship to Perivascular Astrocytic Swelling
  22. The Effect of Continuous Treatment with Sodium Nitroprusside on the Serum Kinetics of the Brain Marker Protein S-100β in Neonates Undergoing Corrective Cardiac Surgery by Means of Hypothermic Cardiopulmonary Bypass
  23. Differentiated Therapy with Prostaglandin E1 (Alprostadil) after Orthotopic Liver Transplantation: the Usefulness of Procalcitonin (PCT) and Hepatic Artery Resistive Index (RI) for the Evaluation of Early Graft Function and Clinical Course
  24. The Postoperative Course of γ-Glutamyl Transpeptidase – a Marker of Cytomegalovirus (CMV) Replication Risk?
  25. TNF-α and Its Receptors Mediate Graft Rejection and Loss after Liver Transplantation
  26. Impact of Ig-Therasorb® Immunoapheresis on Stability of Xenogeneic Ex Vivo Porcine Liver Perfusion – Value of Aminotransferases and Flow Rates for the Assessment of Metabolic Graft Viability
  27. The Effect of Human Organ Preservation and Albumin Flush Solution on In Vitro Cell Metabolic Activity
  28. An Ex Vivo Model of Tolerance vs. Rejection: Comparison of Different Signal Transducers and Activators of Transcription, STAT1, STAT4, STAT5 and STAT6
  29. Detection of Cytomegalovirus (CMV) Antigens in Kidney Biopsies and Transplant Nephrectomies as a Marker for Renal Graft Dysfunction
  30. Comparison of Five Cyclosporin Immunoassays with HPLC
  31. In Vitro Pentamer Formation as a Biomarker of Tacrolimus-Related Immunosuppressive Activity after Liver Transplantation
  32. Pharmacodynamic Monitoring of Mycophenolate Mofetil
https://doi.org/10.1515/CCLM.2000.190

Articles in the same Issue

  1. European Meeting on Biomarkers of Organ Damage and Dysfunction Selected Papers from the Meeting Held in Cambridge, UK April 3rd-7th, 2000
  2. Kidney Retrieval Conditions Influence Damage to Renal Medulla: Evaluation by Proton Nuclear Magnetic Resonance (NMR) Spectroscopy
  3. Citrate, Acetate and Renal Medullary Osmolyte Excretion in Urine as Predictor of Renal Changes after Cold Ischaemia and Transplantation
  4. Comparison of Proteolytic Enzymes and Glutathione S-Transferase Levels in Non-Heart-Beating Donors' (NHBD) Kidney Perfusates
  5. Assessment of Non-Heart-Beating Donor (NHBD) Kidneys for Viability on Machine Perfusion
  6. Low Levels of Urinary Albumin Excretion Are Associated with Cardiovascular Risk Factors in the General Population
  7. Evaluation of a Fully Automated Assay to Measure C-Telopeptide of Type I Collagen in Serum
  8. Bone Turnover Markers and Estradiol Level in Postmenopausal Women
  9. External Quality Assessment of Bone Metabolism Marker Assays. Initial Experiences in a UK NEQAS Programme
  10. The Monoethylglycinexylidide (MEGX) Test as a Marker of Hepatic Dysfunction in Septic Patients with Pneumonia
  11. Transforming Growth Factor-β1 as a Surrogate Marker of Hepatic Dysfunction in Chronic Liver Diseases
  12. A New Approach to the Evaluation of Liver Graft Function by Nuclear Magnetic Resonance Spectroscopy. A Comparative Study between Euro-Collins and University of Wisconsin Solutions
  13. Metallothionein and Heat Shock Protein Expression during Acute Liver Injury and Regeneration in Rats
  14. Biochemical Evaluation of Patients with Acute Pancreatitis
  15. Screening for Acute Myocardial Injury: Creatine Kinase Is Comparable to Myoglobin
  16. Different Time Frames for the Occurrence of Elevated Levels of Cardiac Troponin T and C-Reactive Protein in Patients with Acute Myocardial Infarction
  17. Determination of Decision Limits for ACS:Systems Cardiac Troponin I
  18. Stability of Cardiac Troponins and CK-MB Mass Isoenzyme
  19. Cytokines as Plasma Markers of Abdominal Aortic Aneurysm
  20. The Early Fall in Levels of S-100 β in Traumatic Brain Injury
  21. Protein S-100β in Brain and Serum after Deep Hypothermic Circulatory Arrest in Rabbits: Relationship to Perivascular Astrocytic Swelling
  22. The Effect of Continuous Treatment with Sodium Nitroprusside on the Serum Kinetics of the Brain Marker Protein S-100β in Neonates Undergoing Corrective Cardiac Surgery by Means of Hypothermic Cardiopulmonary Bypass
  23. Differentiated Therapy with Prostaglandin E1 (Alprostadil) after Orthotopic Liver Transplantation: the Usefulness of Procalcitonin (PCT) and Hepatic Artery Resistive Index (RI) for the Evaluation of Early Graft Function and Clinical Course
  24. The Postoperative Course of γ-Glutamyl Transpeptidase – a Marker of Cytomegalovirus (CMV) Replication Risk?
  25. TNF-α and Its Receptors Mediate Graft Rejection and Loss after Liver Transplantation
  26. Impact of Ig-Therasorb® Immunoapheresis on Stability of Xenogeneic Ex Vivo Porcine Liver Perfusion – Value of Aminotransferases and Flow Rates for the Assessment of Metabolic Graft Viability
  27. The Effect of Human Organ Preservation and Albumin Flush Solution on In Vitro Cell Metabolic Activity
  28. An Ex Vivo Model of Tolerance vs. Rejection: Comparison of Different Signal Transducers and Activators of Transcription, STAT1, STAT4, STAT5 and STAT6
  29. Detection of Cytomegalovirus (CMV) Antigens in Kidney Biopsies and Transplant Nephrectomies as a Marker for Renal Graft Dysfunction
  30. Comparison of Five Cyclosporin Immunoassays with HPLC
  31. In Vitro Pentamer Formation as a Biomarker of Tacrolimus-Related Immunosuppressive Activity after Liver Transplantation
  32. Pharmacodynamic Monitoring of Mycophenolate Mofetil
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