Nephrol Dial Transplant (2002) 17: 573-579
© 2002 European Renal Association-European Dialysis and Transplant Association
Rapid Communication
Quantitative gene expression of TGF-ß1, IL-10, TNF-
and Fas Ligand in renal cortex and medulla
1 Department of Paediatric Nephrology and Metabolic Diseases, Children's Hospital, Hannover Medical School, D-30623 Hannover, Germany and 2 Division of Nephrology and Immunology, University of Alberta, 250 Heritage Medical Research Centre, Edmonton, Alberta T6G 2S2, Canada
Abstract
Background. Inflammatory, fibrogenic or apoptotic processes in the kidney are regulated by intra- and intercellular mediators. Intrarenal upregulation of genes may precede structural changes by days and can be examined in extremely small amounts of tissue. With the advent of new quantitative PCR methods results of gene expression are available within few hours after kidney biopsies.
Methods. In order to establish reference values for intrarenal gene expression of TGF-ß1, IL-10, TNF-
and Fas Ligand in renal cortex and medulla, we analysed 28 histologically normal kidney samples available after tumour nephrectomy by quantitative real-time PCR. After reverse transcription of isolated RNA, cDNA aliquots were quantified for target genes using the threshold cycle (Ct) method normalized for the house keeping gene GAPDH.
Results. Expression of target genes was lower in cortex as compared to medulla, but the differences were only significant for IL-10 (P=0.0125). TGF-ß1 was found with the highest gene expression about five PCR-cycles (delta Ct) after GAPDH with markedly lower results for TNF- (delta Ct 
9), IL-10 (12) and Fas Ligand (14).
Conclusions. These results are the first reported reference values for human intrarenal gene expression which should facilitate interpretation of data from native or transplant kidneys in future studies.
Keywords: cytokines; delta delta CT method; intrarenal gene expression; kidney transplantation; quantitative RT-PCR
Notes
Correspondence and offprint requests to: Dr Christian von Schnakenburg, PhD, Kinderklinik, Universitätsklinikum Göttingen, Robert-Koch-Strasse 40, D-37075 Göttingen, Germany. Email: cschnak{at}gwdg.de
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