title: Variation in leukocyte subset concentrations affects calcineurin activity measurement: implications for pharmacodynamic monitoring strategies.
abstract: BACKGROUND
  In renal transplantation patients, therapeutic drug monitoring of the calcineurin (CN) inhibitor cyclosporin A (CsA) is mandatory because of the drug's narrow therapeutic index. Pharmacodynamic monitoring of CN inhibition therapy could provide a tool to define and maintain the therapeutic efficacy of CsA therapy. We investigated the effect of variation in cell counts of leukocyte subsets on leukocyte CN activity measurement in renal transplant recipients.
  
  
  METHODS
  We measured leukocyte CN activity, whole blood CsA concentrations, and leukocyte subset cell counts in 25 renal transplant recipients. Blood was collected before graft implantation and CsA therapy, 1 day before transplantation when CsA therapy was already started, and 5 days after transplantation. Monocyte, granulocyte, CD4+ T-cell, CD8+ T-cell, B-cell, and natural killer-cell CN activities and CsA inhibition sensitivities were determined in vitro by a spectrophotometric CN assay.
  
  
  RESULTS
  Leukocyte CN activity was inhibited after drug intake. Inter- and intrapatient variation in leukocyte subset cell counts resulted in variation of sample composition. The mean (SD) CN activity varied among leukocyte cell subsets, ranging from 650 (230) to 166 (26) pmol/min/10(6) cells for monocytes and CD4+ T cells, respectively. CsA half maximal inhibitory concentration (IC(50)) values ranged from 15 to 78 microg/L for monocytes and B cells, respectively.
  
  
  CONCLUSIONS
  Inter- and intraindividual leukocyte subset cell count variation can affect measured CN activity independent of CsA concentration. Cell-specific activity and drug sensitivity should be considered for sample validation to optimize method specificity when pharmacodynamic monitoring strategies are applied in a clinical setting.
authors.affiliation: Department of Clinical Chemistry, Leiden University Medical Center, Leiden, the Netherlands. H.Rossum@lumc.nl
authors.forename: Huub H
authors.initials: HH
authors.lastname: van Rossum
authors.forename: Fred P H T M
authors.initials: FP
authors.lastname: Romijn
authors.forename: Kathryn J
authors.initials: KJ
authors.lastname: Sellar
authors.forename: Nico P M
authors.initials: NP
authors.lastname: Smit
authors.forename: Paul J M
authors.initials: PJ
authors.lastname: van der Boog
authors.forename: Johan W
authors.initials: JW
authors.lastname: de Fijter
authors.forename: Johannes
authors.initials: J
authors.lastname: van Pelt
mesh_terms: D001402:B-Lymphocytes / Q000378:metabolism / Q000473:pathology; D015704:CD4 Antigens / Q000378:metabolism; D016827:CD8 Antigens / Q000378:metabolism; D019703:Calcineurin / Q000097:blood*; D065095:Calcineurin Inhibitors; D016572:Cyclosporine / Q000097:blood / Q000494:pharmacology*; D016903:Drug Monitoring; D005260:Female; D006098:Granulocytes / Q000378:metabolism / Q000473:pathology*; D006801:Humans; D007166:Immunosuppressive Agents / Q000097:blood / Q000494:pharmacology*; D016030:Kidney Transplantation; D007694:Killer Cells, Natural / Q000378:metabolism / Q000473:pathology*; D016131:Lymphocyte Subsets / Q000378:metabolism / Q000473:pathology*; D008297:Male; D008875:Middle Aged; D009000:Monocytes / Q000378:metabolism / Q000473:pathology*; D013601:T-Lymphocytes / Q000378:metabolism / Q000473:pathology
chemical_list: D015704:CD4 Antigens; D016827:CD8 Antigens; D065095:Calcineurin Inhibitors; D007166:Immunosuppressive Agents; D016572:Cyclosporine; D019703:Calcineurin