title: The pharmacodynamic effect of sirolimus: individual variation of cytokine mRNA expression profiles in human whole blood samples.
abstract: Sirolimus (SRL) has become an important alternative to calcineurin inhibitors due to its unique mechanism of action. Since rejection and poor graft outcome are still frequent problems despite therapeutic-range blood concentrations, pharmacodynamic measurements of its immunosuppressive effects would be of great clinical value to optimize treatment in individual patients. We performed a human whole blood assay using real time cytokine RT-PCR for the pharmacodynamic assessment of SRL. IL-2, IL-4 and IL-6 mRNA levels were quantitatively determined upon T-cell-specific stimulation in healthy individuals (n=11; in vitro) and in kidney-transplant patients (n=3; ex vivo). Furthermore, IL-2 protein secretion and T-cell proliferation was measured. After 24h incubation we observed a stronger suppression of IL-2 and IL-4 mRNA expression upon SRL addition (p<0.005; p<0.005) versus 4h (p<0.05; p<0.05). SRL effects displayed a remarkable interindividual variation, which proved to be independent of the concentration applied. Notably, 3/11 and 2/11 individuals had unaffected IL-2 and IL-4 mRNA expression after 4h incubation with SRL, respectively. In contrast, a general suppression of IL-2 protein secretion and T-cell proliferation was induced. Analysis of kidney-transplant patients verified interindividual variation and proved comparability of in vitro and ex vivo effects. We describe an individual degree of SRL-sensitivity that may correlate with clinical efficacy. Rather than analysis of one single peak, we suggest determination of two absolute cytokine mRNA peak levels for the pharmacodynamic assessment of SRL. However, prospective clinical studies are necessary to determine whether individual degrees of SRL-sensitivity correlate with clinical outcome.
authors.affiliation: Institute of Transfusion Medicine and Immunology, Faculty of Clinical Medicine Mannheim, University of Heidelberg, Friedrich-Ebert-Strasse 107, 68167 Mannheim, Germany. m.mueller-steinhardt@blutspende.de
authors.forename: Michael
authors.initials: M
authors.lastname: Müller-Steinhardt
authors.forename: Kristina
authors.initials: K
authors.lastname: Wortmeier
authors.forename: Lutz
authors.initials: L
authors.lastname: Fricke
authors.forename: Brigitte
authors.initials: B
authors.lastname: Ebel
authors.forename: Christoph
authors.initials: C
authors.lastname: Härtel
mesh_terms: D000368:Aged; D017252:CD3 Complex / Q000276:immunology; D049109:Cell Proliferation / Q000187:drug effects; D020869:Gene Expression Profiling; D006084:Graft Rejection / Q000097:blood / Q000235:genetics / Q000378:metabolism* / Q000517:prevention & control*; D006801:Humans; D007165:Immunosuppression Therapy; D007166:Immunosuppressive Agents / Q000493:pharmacokinetics*; D007376:Interleukin-2 / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D015847:Interleukin-4 / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D015850:Interleukin-6 / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D016030:Kidney Transplantation / Q000276:immunology*; D008213:Lymphocyte Activation / Q000187:drug effects; D008875:Middle Aged; D012333:RNA, Messenger / Q000032:analysis*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D020123:Sirolimus / Q000493:pharmacokinetics*; D013601:T-Lymphocytes / Q000276:immunology / Q000378:metabolism* / Q000473:pathology
chemical_list: D017252:CD3 Complex; D007166:Immunosuppressive Agents; D007376:Interleukin-2; D015850:Interleukin-6; D012333:RNA, Messenger; D015847:Interleukin-4; D020123:Sirolimus