Related Experiment Video
Updated: Aug 30, 2026
Murine Kidney Transplant Technique
Published on: October 20, 2015
Cytokine gene expression in kidney allograft donor biopsies after cold ischemia and reperfusion using in situ RT-PCR
D Kaminska1, B Tyran, O Mazanowska
1Department of Nephrology and Transplantation Medicine, Wroclaw Medical University, Poland. dorotakaminska@interia.pl
Abstract:
It was previously reported that ischemia-reperfusion injury initiates an inflammatory response and may significantly affect the transplanted organ function. The aim of this study was to assess changes of intragraft cytokine mRNA expression in kidneys after cold ischemia (CI) and following reperfusion. We examined mRNA of a product of activated T lymphocytes (IFN-gamma) and a monocyte product (IL-6). Eleven kidneys were transplanted after CI time ranging from 16 to 39 hours. Renal needle core biopsies were obtained from donors after cold ischemia and approximately after 20 minutes of reperfusion. Tubular and glomerular expression of IFN-gamma and IL-6 mRNA were assessed using semiquantitative evaluation of the RT-PCR in situ. After reperfusion an intense increase of IL-6 mRNA expression was observed in four specimens, a slight increase was noticed in five specimens, and a very slight decrease in two specimens. Changes in IL-6 mRNA expression were limited only to tubules. In contrast, the glomerular and tubular mRNA expression of IFN-gamma and glomerular of IL-6 remained stable. Mean CI time for patients with an intense increase was higher than for patients with a slight increase and with the decrease of IL-6 mRNA expression (32.0 +/- 6.8 vs 25.2 +/- 7.3 and 26.0 +/- 5.7 hours). Our results suggest that early inflammatory changes at the time of implantation of renal allografts depends mainly on monocyte/macrophage-associated products. The observed intensity of their expression in tubules was connected to longer CI time.
Insights
Cold ischemia and reperfusion in kidney transplants increase interleukin-6 (IL-6) mRNA expression in tubules, linked to longer cold ischemia times. Interferon-gamma (IFN-gamma) expression remained stable, suggesting early inflammation involves monocyte products.
Area of Science:
- Transplantation immunology
- Renal pathology
- Molecular biology
Background:
- Ischemia-reperfusion injury (IRI) is a critical factor impacting transplanted organ function.
- Early inflammatory responses post-transplantation can significantly affect graft outcomes.
- Understanding intragraft molecular changes is crucial for mitigating IRI.
Purpose of the Study:
- To investigate changes in intragraft cytokine messenger RNA (mRNA) expression in kidneys following cold ischemia (CI) and reperfusion.
- To specifically assess the expression of interferon-gamma (IFN-gamma) and interleukin-6 (IL-6) mRNA in renal allografts.
Main Methods:
- Analysis of renal needle core biopsies from eleven kidney transplant donors.
- Assessment of IFN-gamma and IL-6 mRNA expression in tubular and glomerular compartments using in situ RT-PCR.
- Evaluation of expression levels post-cold ischemia and after reperfusion.
Main Results:
- A significant increase in IL-6 mRNA expression was observed in tubules post-reperfusion in most specimens.
- IFN-gamma mRNA expression (tubular and glomerular) and glomerular IL-6 mRNA expression remained stable.
- Higher mean CI times correlated with more intense IL-6 mRNA upregulation in tubules.
Conclusions:
- Early inflammatory changes in renal allografts are primarily associated with monocyte/macrophage-derived products like IL-6.
- The intensity of IL-6 expression in tubules is linked to the duration of cold ischemia.
- Targeting early inflammatory mediators may be beneficial in reducing IRI severity.
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