Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
3392506 | Transplant Immunology | 2009 | 4 Pages |
IntroductionInitial high dose glucocorticosteroid (GC) treatment of transplant recipients induces changes in the glucocorticoid receptor (GR) that may affect the mode of action of the drug. We developed a TaqMan one-step RT-PCR quantification method to investigate the effect of bolus GC treatment on the structure of the GR complex by measuring the levels of GR isoform expression and FK binding protein 5 (FKBP5) transcription.MethodsPeripheral blood mononuclear cells (PBMCs) were obtained before renal transplantation and immediately after the initial methylprednisolone treatment. Gene expression of the GR-α, -β, -P isoforms and FKBP5 were quantified using a simplex TaqMan relative quantification assay. Genotyping was performed using the TaqMan allele discrimination assay.ResultsThe gene expression assay was validated and is efficient to detect extremely low quantity of GR-β expression. Increased expression of FKBP5 (14.01 + 11.52 folds), GR-α (1.47 + 1.00 folds), GR-β (6.21 + 5.71 folds) and GR-P (2.72 + 1.90 folds) were observed after steroid bolus. The GR haplotype A-T-G is significantly related to elevation of GRβ transcription (p = 0.002). Patients’ hospitalization time after transplantation correlated with increment of GR-α (p = 0.01) and FKBP5 (p = 0.007) gene expression. The GR-β expression level is extremely low compared with the GR-α isoform. By contrast, the GR-P isoform is relatively abundantly expressed in PBMCs.ConclusionThe method to measure GR isoform enabled us to determine that acute high-dose GC treatment alters the structure of the GR. The initial therapy with high dose steroid may induce a GC resistance phenotype. The modulatory role of the GR-P isoform is not fully defined but may provide another dimension in understanding the function of GR and the role of steroid in transplant immunosuppression.