PURPOSE: The purpose of the study was to determine if cytokines could be measured from the supernatant of kidney biopsy cell suspensions and if measured cytokines correlated with tissue pathology and clinical course.

METHODS: In preparing cell suspensions for flow cytometry from one third of a 16 gauge renal biopsy core, a cell-free supernatant is obtained. We used the Milliplex high sensitivity bead capture and Luminex platform to determine the amounts of IL-1-beta, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12/p70, IL-13, IFN-gamma, GM-CSF and TNF-alpha in this supernatant. Cytokine levels can be normalized to the total number of cells associated with the supernatant.

RESULTS: Cytokines within renal biopsies are reproducibly measureable at picogram/ml levels, which is possible with the use of a high-sensitivity Milliplex system. IL-6 and IL-8 were elevated in kidneys with recent ischemia. TNF-alpha was highest in kidneys with moderate-severe persistent rejection. Compared to normal native and transplanted kidneys, higher IL-4 and IL-10 were found in allografts with rejection and nonspecific inflammation. IL-5 and GM-CSF were not detected in any of the 150 kidneys assessed for the presence of cytokines. IL-2, IFN-gamma, IL-1-beta, IL-12/p70 and IL-13 levels were detected at low levels and did not have an obvious correlation with pathology or clinical course.

CONCLUSION: Cytokines can be measured within kidney allograft biopsies and levels vary with pathology. This provides useful information about the condition of allografts without requiring more tissue than is standardly obtained for transplant kidney biopsies.

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