title: Validation of immunological biomarkers for the pharmacodynamic monitoring of immunosuppressive drugs in humans.
abstract: Pharmacodynamic monitoring (PD) can evaluate the efficacy of immunosuppressive drug therapies. In this study, the expressions of PD biomarkers [lymphocyte proliferation, CD25 and CD71 expression, interleukin-2 (IL-2), and tumor necrosis factor-alpha (TNF-alpha) synthesis] were determined in whole-blood assays and were validated for their application in PD of immune modulators in future clinical trials. Initially, the assay conditions were re-evaluated. The measurement of T-lymphocyte proliferation and activation marker expression in whole-blood cultures resulted in optimized stimulation for 72 hours with 7.5 microg/mL concavalin A. Intracellular cytokine expression of CD3+ T-cells received optimized stimulation for 4 hours with 15 ng/mL phorbol 12-myristate 13-acetate and 0.75 microg/mL ionomycin. Statistical assay parameters (intra-assay, intra-individual, and interindividual variabilities) were determined. It was found that blood storage for up to 24 hours is possible without any change in biomarker expression. Dosage effects of immunosuppressive drugs (tacrolimus, cyclosporin A, sirolimus, mycophenolic acid, and methylprednisolone) were evaluated in vitro and the assay was applied successfully to dialysis, renal transplant, and liver transplant patients. We conclude that these biomarkers used in whole-blood assays are suitable for PD of immune modulators in clinical trials.
authors.affiliation: INSERM U466, Institut Louis Bugnard, Toulouse, France. boehlert@toulouse.inserm.fr
authors.forename: Torsten
authors.initials: T
authors.lastname: Böhler
authors.forename: Jochen
authors.initials: J
authors.lastname: Nolting
authors.forename: Nassim
authors.initials: N
authors.lastname: Kamar
authors.forename: Purevtseren
authors.initials: P
authors.lastname: Gurragchaa
authors.forename: Kathrin
authors.initials: K
authors.lastname: Reisener
authors.forename: Petra
authors.initials: P
authors.lastname: Glander
authors.forename: Hans-H
authors.initials: HH
authors.lastname: Neumayer
authors.forename: Klemens
authors.initials: K
authors.lastname: Budde
authors.forename: Jochen
authors.initials: J
authors.lastname: Klupp
mesh_terms: D000328:Adult; D015703:Antigens, CD / Q000097:blood; D015415:Biomarkers / Q000097:blood*; D015496:CD4-Positive T-Lymphocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D049109:Cell Proliferation / Q000187:drug effects; D003208:Concanavalin A / Q000494:pharmacology; D016572:Cyclosporine / Q000494:pharmacology; D016903:Drug Monitoring / Q000379:methods*; D006801:Humans; D007166:Immunosuppressive Agents / Q000145:classification / Q000494:pharmacology*; D007376:Interleukin-2 / Q000097:blood; D053645:Interleukin-2 Receptor alpha Subunit / Q000097:blood; D008213:Lymphocyte Activation / Q000187:drug effects; D008775:Methylprednisolone / Q000494:pharmacology; D008875:Middle Aged; D009173:Mycophenolic Acid / Q000494:pharmacology; D011990:Receptors, Transferrin / Q000097:blood; D015203:Reproducibility of Results; D020123:Sirolimus / Q000494:pharmacology; D016559:Tacrolimus / Q000494:pharmacology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014409:Tumor Necrosis Factor-alpha / Q000097:blood
chemical_list: D015703:Antigens, CD; D015415:Biomarkers; C068126:CD71 antigen; D007166:Immunosuppressive Agents; D007376:Interleukin-2; D053645:Interleukin-2 Receptor alpha Subunit; D011990:Receptors, Transferrin; D014409:Tumor Necrosis Factor-alpha; D003208:Concanavalin A; D016572:Cyclosporine; D009173:Mycophenolic Acid; D013755:Tetradecanoylphorbol Acetate; D020123:Sirolimus; D016559:Tacrolimus; D008775:Methylprednisolone