Analytical Validation and Cross-Validation of an NFAT-Regulated Gene Expression Assay for Pharmacodynamic Monitoring of Therapy With Calcineurin Inhibitors.


BACKGROUND:Analysis of residual gene expression of the nuclear factor of activated T cell (NFAT)-regulated genes has been developed as a pharmacodynamic biomarker to monitor therapy with calcineurin inhibitors. The availability of commercial primer sets (Search-LC) and the well-established assay protocol makes this biomarker a promising candidate to be used clinically in different laboratories. However, implementation of the method in routine practice requires analytical robustness and comparable results across laboratories. Therefore, a protocol originally established at the Institute of Immunology, Heidelberg was verified at the Institute of Laboratory Medicine, Klinikum Stuttgart, and a comparison study was conducted between the 2 laboratories. METHODS:For the analytical verification, whole blood samples of healthy individuals were incubated with tacrolimus in vitro. Linearity, imprecision, and limit of quantification, as well as sample stability, were investigated. For interlaboratory comparison, samples of patients under cyclosporine A therapy were analyzed in Heidelberg and then reanalyzed in Stuttgart within 24 hours. RESULTS:Tacrolimus (6.25-50 mcg/L) decreased the expression of NFAT-regulated genes in vitro dose dependently (15%-89%). Within- and between-assay coefficient of variations (n = 6 each) were <17%. The limit of quantification was <200 cDNA copies for each of the interleukin-2, interferon-γ, and granulocyte-macrophage colony-stimulating factor genes. Samples were stable for 24 hours. Interlaboratory comparison using patient samples correlated well (r = 0.951) but showed an inconsistent bias depending on the magnitude of residual gene expression. CONCLUSIONS:The assay can be set up with a satisfactory analytical performance in a routine molecular biological laboratory and shows comparable results between laboratories. The reproducibility of the NFAT-regulated gene expression assay across laboratories can facilitate the implementation of this assay for pharmacodynamic routine monitoring of calcineurin inhibitors in different centers.


Ther Drug Monit


Abdel-Kahaar E,Giese T,Sommerer C,Rieger H,Shipkova M,Wieland E




Has Abstract


2016-12-01 00:00:00












  • Determinants of mycophenolic acid levels after renal transplantation.

    abstract::There are data suggesting an association between mycophenolic acid (MPA) levels and acute rejection and toxicity in renal transplant recipients treated with mycophenolate mofetil (MMF), and therefore, knowledge of factors determining MPA levels may aid in accurate adjustment of MMF dosage. A total of 4970 samples take...

    journal_title:Therapeutic drug monitoring

    pub_type: 临床试验,杂志文章


    authors: Borrows R,Chusney G,James A,Stichbury J,Van Tromp J,Cairns T,Griffith M,Hakim N,McLean A,Palmer A,Papalois V,Taube D

    更新日期:2005-08-01 00:00:00

  • Δ9-Tetrahydrocannabinol and Cannabidiol Time Courses in the Sera of "Light Cannabis" Smokers: Discriminating Light Cannabis Use from Illegal and Medical Cannabis Use.

    abstract:BACKGROUND:Δ9-tetrahydrocannabinol (THC) and cannabidiol (CBD) time courses in serum and physiological and behavioral effects associated with smoking 1 or 4 "light cannabis" cigarettes were studied. Biomarkers to differentiate light cannabis versus illegal and medical cannabis use were also investigated. METHODS:Sera ...

    journal_title:Therapeutic drug monitoring

    pub_type: 杂志文章


    authors: Pichini S,Mannocchi G,Berretta P,Zaami S,Pirani F,Pacifici R,Busardò FP

    更新日期:2020-02-01 00:00:00

  • Monitoring cyclosporin in blood: between-assay differences at trough and 2 hours post-dose (C2).

    abstract::With the introduction of a cyclosporin monitoring strategy based on the use of a sample collected 2 hours after dosing (C2) rather than the predose sample (C0), there was concern that the differences in blood cyclosporin results from the various assay systems would result in assay-specific target ranges for C2 monitor...

    journal_title:Therapeutic drug monitoring

    pub_type: 临床试验,杂志文章,多中心研究


    authors: Johnston A,Chusney G,Schütz E,Oellerich M,Lee TD,Holt DW

    更新日期:2003-04-01 00:00:00

  • Old and new antirheumatic drugs and the risk of hepatotoxicity.

    abstract::Given the high prevalence of the use of medications in daily practice and the large number of people taking antirheumatic agents, the risk of drug-drug interactions and of hepatotoxicity is of concern. Both old and new compounds show such a risk. Nonsteroidal antinflammatory drugs are widely used drugs with potential ...

    journal_title:Therapeutic drug monitoring

    pub_type: 杂志文章,评审


    authors: Anelli MG,Scioscia C,Grattagliano I,Lapadula G

    更新日期:2012-12-01 00:00:00

  • Cancer Chemotherapy: The Case for Therapeutic Drug Monitoring.

    abstract::The selection of an appropriate therapy and dosing regimen is a significant challenge in the treatment of cancer. Although there are recommended standardized chemotherapy protocols for some types of cancer, protocol changes that usually only occur after large clinical trials demonstrate improvements and individual pat...

    journal_title:Therapeutic drug monitoring

    pub_type: 杂志文章


    authors: Knezevic CE,Clarke W

    更新日期:2020-02-01 00:00:00

  • Monitoring phenytoin in salivary and plasma ultrafiltrates of pediatric patients.

    abstract::The plasma binding of phenytoin and the relationship between phenytoin concentrations in salivary and plasma ultrafiltrates were evaluated in pediatric epileptic patients aged 2-15 years. Paired samples of plasma and saliva were ultrafiltered through an Amicon YMT membrane. Phenytoin concentrations were measured by a ...

    journal_title:Therapeutic drug monitoring

    pub_type: 杂志文章


    authors: Bachmann K,Forney RB Jr,Voeller K

    更新日期:1983-01-01 00:00:00

  • Plasma concentrations of dothiepin and its metabolites are not correlated with clinical efficacy in major depressive illness.

    abstract::Plasma concentration-antidepressant response relationships for dothiepin, nordothiepin, dothiepin-S-oxide, and nordothiepin-S-oxide were investigated in 50 patients (33 women and 17 men), who had had a major depressive episode. Depression and anxiety were assessed at the start of therapy and after 2 and 4 weeks by mea...

    journal_title:Therapeutic drug monitoring

    pub_type: 临床试验,杂志文章


    authors: Ilett KF,Blythe TH,Hackett LP,Ong RT,Tannenbaum DA,Clarke TM

    更新日期:1993-10-01 00:00:00

  • Evaluation of Infliximab and Anti-infliximab LISA-TRACKER Immunoassays for the Therapeutic Drug Monitoring of SB2 Infliximab Biosimilar.

    abstract:BACKGROUND:SB2, an infliximab (IFX) biosimilar to the reference infliximab (R.I.) product (Remicade), received approval in the European Union for all IFX indications. Many decision algorithms based on the measurement of IFX trough levels and antibodies to infliximab are being increasingly used to optimize IFX treatment...

    journal_title:Therapeutic drug monitoring

    pub_type: 杂志文章


    authors: Jentzer A,Berger AE,Labetoulle R,Haccourt A,Roblin X,Paul S

    更新日期:2018-12-01 00:00:00

  • Oral aripiprazole-induced severe hypoglycemia.

    abstract::This case report highlights a very rare adverse drug reaction caused by oral aripiprazole resulting in severe hypoglycemia. A 72-year-old-male patient suffering from Parkinson disease on prolonged carbidopa plus levodopa combination therapy (carbidopa 25 mg plus levodopa 100 mg, thrice daily) for 1.3 years was recentl...

    journal_title:Therapeutic drug monitoring

    pub_type: 杂志文章,评审


    authors: Mondal S,Saha I,Das S,Ganguly A,Ghosh A,Kumar Das A

    更新日期:2012-06-01 00:00:00

  • Comparison of particle-enhanced turbidimetric inhibition immunoassay and fluorescence polarization immunoassay using monoclonal antibodies for theophylline.

    abstract::The DuPont theophylline assay reagent kit, a particle-enhanced turbidimetric inhibition immunoassay (PETINIA) method adapted for use on a centrifugal fast analyzer, was evaluated. It was compared with fluorescence polarization immunoassay (FPIA). Day-to-day precision was 4.7% at 6.8 micrograms/ml, and 3.3% at 26.6 mic...

    journal_title:Therapeutic drug monitoring

    pub_type: 杂志文章


    authors: Lawrence JL,Elser RC

    更新日期:1986-01-01 00:00:00

  • Multicenter quality control study of amikacin assay for monitoring once-daily dosing regimens. International Antimicrobial Therapy Cooperative Group of the European Organization for Research and Treatment of Cancer.

    abstract::During once-daily dosing regimens of aminoglycosides, administration of large single doses results in high peak levels and low 24-h trough levels. However, commercial assays for monitoring aminoglycoside levels are designed to cover the smaller range of serum concentrations usually observed during multiple daily dosin...

    journal_title:Therapeutic drug monitoring

    pub_type: 临床试验,杂志文章,多中心研究


    authors: Blaser J,König C,Fatio R,Follath F,Cometta A,Glauser M

    更新日期:1995-04-01 00:00:00

  • Pharmacokinetics of high-dose oral and intravenous dexamethasone.

    abstract::Pharmacokinetics of intravenous and oral pulsed high-dose dexamethasone were studied in four patients with pemphigus vulgaris. Doses for dexamethasone were varied from 100 to 300 mg. Serum concentrations were measured by high-performance liquid chromatographic procedure with diode assay detection. Bioavailability was ...

    journal_title:Therapeutic drug monitoring

    pub_type: 杂志文章


    authors: Tóth GG,Kloosterman C,Uges DR,Jonkman MF

    更新日期:1999-10-01 00:00:00

  • Multicenter analytical evaluation of the automated electrochemiluminescence immunoassay for cyclosporine.

    abstract:BACKGROUND:Cyclosporine A (CsA) is used as a posttransplantation immunosuppressant drug, and careful monitoring of CsA concentration in whole blood is essential. A new automated electrochemiluminescence immunoassay (ECLIA) for CsA measurement has been assessed in a multicenter evaluation. METHODS:Residual EDTA whole b...

    journal_title:Therapeutic drug monitoring

    pub_type: 杂志文章,多中心研究


    authors: Vogeser M,Shipkova M,Rigo-Bonnin R,Wallemacq P,Orth M,Widmann M,Verstraete AG

    更新日期:2014-10-01 00:00:00

  • Acute antidepressant response and plasma levels of bupropion and metabolites in a pediatric-aged sample: an exploratory study.

    abstract::Studies examining associations between antidepressant response and plasma levels of bupropion and its metabolites have yielded contradictory findings. There have been no such studies in youth. This study explored such associations in 8 boys and 8 girls, age 11 to 17 years, all prescribed bupropion sustained release (S...

    journal_title:Therapeutic drug monitoring

    pub_type: 临床试验,杂志文章


    authors: Daviss WB,Perel JM,Brent DA,Axelson DA,Rudolph GR,Gilchrist R,Nuss S,Birmaher B

    更新日期:2006-04-01 00:00:00

  • Possible interaction between cyclosporine and glibenclamide in posttransplant diabetic patients.

    abstract::The possible occurrence of a kinetic interaction between cyclosporine A and glibenclamide was assessed by reviewing data of six posttransplant diabetic patients who received the two drugs concurrently. Coadministration of the two drugs resulted in a 57% increase in the steady-state plasma cyclosporine levels despite n...

    journal_title:Therapeutic drug monitoring

    pub_type: 杂志文章


    authors: Islam SI,Masuda QN,Bolaji OO,Shaheen FM,Sheikh IA

    更新日期:1996-10-01 00:00:00

  • A rapid and simple microassay for lidocaine in human blood plasma using gas-liquid chromatography with nitrogen detection.

    abstract::Lidocaine is widely used in antiarrhythmic therapy, and the need to monitor the drug concentration in the blood of patients receiving lidocaine is well established. We have developed a new assay for lidocaine designed for the routine clinical therapeutic drug monitoring laboratory. A 100-microliter aliquot of blood pl...

    journal_title:Therapeutic drug monitoring

    pub_type: 杂志文章


    authors: Gal J,Freedman MD,Kumar E,Freed CR

    更新日期:1981-01-01 00:00:00

  • External evaluation of population pharmacokinetics and pharmacodynamics in linezolid-induced thrombocytopenia: the transferability of published models to different hospitalized patients.

    abstract:OBJECTIVES:The objective of the present study was to perform an external evaluation of published linezolid population pharmacokinetic and pharmacodynamic models, in order to evaluate the predictive performance using an independent dataset. Another aim was to offer an elegant environment for display and simulation of bo...

    journal_title:Therapeutic drug monitoring

    pub_type: 杂志文章


    authors: Ogami C,Tsuji Y,Nishi Y,Kawasuji H,To H,Yamamoto Y

    更新日期:2020-09-28 00:00:00

  • The effect of hyperlipidemia on therapeutic drug assays.

    abstract::Hyperlipidemia interferes with spectrophotometric assays of a number of analytes. No interference was found in the assay of gentamicin, phenytoin, or phenobarbital due to turbidity in levels of triglyceride less than or equal to 2,000 mg/dl with the Abbott TDx or Syva enzyme multiplied immunoassay (EMIT) methods. Ther...

    journal_title:Therapeutic drug monitoring

    pub_type: 杂志文章


    authors: Baer DM,Paulson RA

    更新日期:1987-01-01 00:00:00

  • Optimum dosage regimen of palivizumab?

    abstract::Palivizumab is a humanized, monoclonal antibody used to protect at-risk infants against respiratory syncytial virus (RSV) infection. The regular dosage scheme causes a low initial trough level and accumulation of the antibody after subsequent injections. Using a simple pharmacokinetic model, the authors devised an alt...

    journal_title:Therapeutic drug monitoring

    pub_type: 杂志文章


    authors: Zaaijer HL,Vandenbroucke-Grauls CM,Franssen EJ

    更新日期:2002-06-01 00:00:00

  • Assessment of cyclosporin A in whole blood and plasma in five patients with different hematocrits.

    abstract::Measurement of whole blood and plasma levels of cyclosporin A (CsA) by radioimmunoassay in kidney transplant recipients receiving the drug showed that CsA concentrations in plasma increased nonlinearly when whole blood levels of the drug exceeded 1,000 ng/ml. At low plasma levels (less than 200 ng/ml), most of the CsA...

    journal_title:Therapeutic drug monitoring

    pub_type: 杂志文章


    authors: Agarwal RP,McPherson RA,Threatte GA

    更新日期:1985-01-01 00:00:00

  • Sorption of thiotepa to polyurethane catheter causes falsely elevated plasma levels.

    abstract::Central venous access catheters are commonly used in clinical oncology. The double lumen variant is applied in pharmacokinetic studies for simultaneous administration and blood sampling when frequent blood collections are necessary. Occlusion of one lumen, a common complication, necessitates the investigator perform b...

    journal_title:Therapeutic drug monitoring

    pub_type: 杂志文章


    authors: de Jonge ME,Mathôt RA,van Dam SM,Rodenhuis S,Beijnen JH

    更新日期:2003-06-01 00:00:00

  • Pharmacokinetic/pharmacodynamic modeling of psychomotor impairment induced by oral clonazepam in healthy volunteers.

    abstract::This study was undertaken to model the relationship between clonazepam plasma concentrations and a central nervous system adverse effect (impairment of the psychomotor performance) following the oral administration of immediate-release tablets of clonazepam in healthy volunteers. Such a (P)pharmacokinetic/(P)pharmacod...

    journal_title:Therapeutic drug monitoring

    pub_type: 杂志文章,随机对照试验


    authors: dos Santos FM,Gonçalves JC,Caminha R,da Silveira GE,Neves CS,Gram KR,Ferreira CT,Jacqmin P,Noël F

    更新日期:2009-10-01 00:00:00

  • Serum levels of olanzapine and its N-desmethyl and 2-hydroxymethyl metabolites in child and adolescent psychiatric disorders: effects of dose, diagnosis, age, sex, smoking, and comedication.

    abstract::The aim of this study was to assess dose-related steady-state serum concentrations of olanzapine (OLZ) and its metabolites N-desmethyl OLZ (DMO) and 2-hydroxymethyl OLZ (2-OH-OLZ) (assessed by high-performance liquid chromatography) in 122 child and adolescent psychiatric patients (age 16.9 +/- 2.2, range, 10-21 years...

    journal_title:Therapeutic drug monitoring

    pub_type: 杂志文章


    authors: Theisen FM,Haberhausen M,Schulz E,Fleischhaker C,Clement HW,Heinzel-Gutenbrunner M,Remschmidt H

    更新日期:2006-12-01 00:00:00

  • The relation between dosage, serum concentrations, and clinical outcome in children and adolescents treated with sertraline: a naturalistic study.

    abstract:OBJECTIVE:This naturalistic therapeutic drug monitoring (TDM) study aimed to evaluate the relationship between dosage, serum concentration, and clinical outcome in children and adolescents treated with the serotonin reuptake inhibitor sertraline for different indications. METHODS:Steady-state trough serum concentratio...

    journal_title:Therapeutic drug monitoring

    pub_type: 杂志文章


    authors: Taurines R,Burger R,Wewetzer C,Pfuhlmann B,Mehler-Wex C,Gerlach M,Egberts K

    更新日期:2013-02-01 00:00:00

  • Carbamazepine and its epoxide: an open study of efficacy and side effects after carbamazepine dose increment in refractory partial epilepsy.

    abstract::We evaluated the efficacy, development of adverse effects, and possible correlation between the plasma concentration of carbamazepine (CBZ) and its major metabolite, carbamazepine-10,11-epoxide (CBZ-E), in a group of epileptic patients in whom selective increases in CBZ doses were made. Eighteen patients with refracto...

    journal_title:Therapeutic drug monitoring

    pub_type: 临床试验,杂志文章


    authors: Semah F,Gimenez F,Longer E,Laplane D,Thuillier A,Baulac M

    更新日期:1994-12-01 00:00:00

  • Azathioprine Therapy in a Pediatric TPMT-Deficient Patient-Still an Option.

    abstract::We describe the case of a pediatric patient on azathioprine therapy with previously undiagnosed homozygote thiopurine S-methyltransferase (TPMT) deficiency, resulting in myelotoxic thiopurine metabolite levels. The patient was successfully treated with a very low azathioprine dose of 50 mg once a week (4% of standard ...

    journal_title:Therapeutic drug monitoring

    pub_type: 杂志文章


    authors: van Moorsel SA,Bevers N,Meurs M,van Rossum LK,Hooymans PM,Wong DR

    更新日期:2017-02-01 00:00:00

  • Mechanisms of action and overview of OKT3.

    abstract::OKT3 is a murine monoclonal antibody of the immunoglobulin IgG2a isotype. The target of OKT3, CD3, is a 17-20 kilodalton (kD) molecule that is part of a multimolecular complex found only on mature T cells and medullary thymocytes. This complex is uniquely situated next to the T-cell receptor for antigen. An interactio...

    journal_title:Therapeutic drug monitoring

    pub_type: 杂志文章,评审


    authors: Norman DJ

    更新日期:1995-12-01 00:00:00

  • Nonparametric estimation of population characteristics of the kinetics of lithium from observational and experimental data: individualization of chronic dosing regimen using a new Bayesian approach.

    abstract::A population analysis of the kinetics of lithium was performed from experimental and observational data in 113 subjects in order to propose a new approach for lithium dosage individualization. The kinetics of lithium is described by a two-compartment model. Age, body weight, height, and serum creatinine are included a...

    journal_title:Therapeutic drug monitoring

    pub_type: 杂志文章


    authors: Taright N,Mentré F,Mallet A,Jouvent R

    更新日期:1994-06-01 00:00:00

  • Pharmacokinetic modeling and development of Bayesian estimators in kidney transplant patients receiving the tacrolimus once-daily formulation.

    abstract:BACKGROUND:The once-daily formulation of tacrolimus has been reported to exhibit the same efficacy and safety profile as compared with the immediate-release form administered twice daily. However, as a result of differences in their pharmacokinetic (PK) profile, the PK models or Bayesian estimators (MAP-BE) previously ...

    journal_title:Therapeutic drug monitoring

    pub_type: 杂志文章


    authors: Saint-Marcoux F,Debord J,Undre N,Rousseau A,Marquet P

    更新日期:2010-04-01 00:00:00

  • Measurement of Non-Vitamin K Antagonist Oral Anticoagulants in Patient Plasma Using Heptest-STAT Coagulation Method.

    abstract:BACKGROUND:Non-vitamin K antagonist oral anticoagulants (NOACs) are approved for several indications for prophylaxis of thromboembolism at fixed oral doses. The analysis of NOAC activity/concentration may be required in special patient populations. Heptest coagulation assay determines both factor Xa and thrombin inhibi...

    journal_title:Therapeutic drug monitoring

    pub_type: 杂志文章


    authors: Du S,Harenberg J,Krämer S,Krämer R,Wehling M,Weiss C

    更新日期:2015-06-01 00:00:00