The Anti‐Inflammatory Effect of Normothermic Machine Perfused Human Kidneys on Monocyte‐Derived Dendritic Cells
Laura W. D. Knijff, Asel S. Arykbaeva, Sandra W. van der Kooij, Dorottya K. de Vries, Mieke F. van Essen, Ian P. J. Alwayn, Henri G. D. Leuvenink, Rutger J. Ploeg, Cees van KootenABSTRACT
Background
The immunomodulatory activity of normothermic machine perfusion (NMP) on donor organs may be reflected by the release of immune‐regulating factors into the perfusate. Dendritic cells (DCs) are potent antigen‐presenting cells and are able to sense these factors. We hypothesized that activation of DCs could be used to monitor the release of these mediators into the perfusate.
Methods
Human monocytes from buffy coats were differentiated into monocyte‐derived DCs (moDCs). Perfusate was obtained from human donor kidneys deemed unsuitable for transplantation ( n = 15) that underwent 6 h of NMP. MoDCs were incubated for 24 h with perfusate collected at different timepoints. DC supernatant was analyzed with Luminex and ELISA. MoDCs were analyzed with flow cytometry for maturation markers and co‐cultured with allogenic T cells.
Results
Incubation with perfusate resulted in moDCs with a reduced surface expression of CD86, without affecting other maturation markers. Analysis of DC supernatant showed there was a specific increase of IL‐10 production upon incubation with perfusate. As a consequence, moDCs incubated with perfusate showed lower T cell stimulatory capacity.
Conclusion
This study showed that the activation status of DCs can be used to monitor the release of factors into the perfusate during NMP with an anti‐inflammatory DC phenotype and function. It demonstrates the feasibility of a cell‐based model to determine immune regulatory effects during ex vivo organ perfusion.