Nephrol Dial Transplant (1994) 9: 599-606
© 1994 European Renal Association-European Dialysis and Transplant Association
research-article
Interleukin 6 production by human proximal tubular epithelial cells in vitro: analysis of the effects of interleukin-1
(IL-1) and other cytokines
1Department of Nephrology, University Hospital Leiden, The Netherlands 2Department of Medicine, Wilford Hall USAF MC, Texas USA
Correspondence and offprint requests to: Correspondence and offprint requests to: Dr B. A. Yard, Dept of Nephrology, University Hospital Leiden, Building 1 C3-p, Rijnsburgerweg 10, 2333 AA Leiden, The Netherland
Proximal tubular epithelial cells (PTEC) from human renal tissue obtained from biopsy or nephrectomy were grown in monoculture and evaluated in vitro at passage 2–4 for interleukin 6 (IL-6) production in response to medium alone or to interleukin 1 alpha (IL-1
), tumour necrosis factor alpha (TNF), interleukin 2 (IL-2), interferon gamma (INF
) or lipopolysaccharide (LPS). IL-6 bioactivity was quantitated using the IL-6-dependent murine hybridoma cell line (B9) and expressed as IL-6 units/ml/105 PTEC. PTEC cell lines exposed to medium alone produced intermediate amounts of IL-6 with substantial variability between cell lines. Introduction of IL-1 resulted in a dose- and time-dependent increase in IL-6 production by PTEC that was maximal at 1 ng/ml IL-1 at 24 h. All PTEC cell lines showed an increased IL-6 production on exposure to IL-1 varying from 1.3- to 24-fold increase over baseline production. This response was completely blocked by anti-rIL-1. No significant IL-6 production by PTEC could be induced by TNF, IL-2, IFN, or LPS over a broad dosage range. Cycloheximide inhibited IL-6 production without irreversible cell toxicity, indicating de-novo synthesis. IL-6 produced by PTEC had a molecular weight of 26-29 kDa as demonstrated by Western blot analysis. Using PCR analysis we could demonstrate upregulation by IL-1 of IL-6 mRNA in a dose-response fashion, indicating that IL-1 regulates IL-6 production at a pretranslational value of protein synthesis. These results show that human cultured PTEC produce IL-6 under both normal and IL-1-stimulated conditions, and suggest that they may have a regulatory function in response to cytokines in the setting of inflammation in the renal cortex.
Keywords: Interleukin 6; epithelial cells; Interleukin-1; proximal tubule
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