Interleukin 6 production by human proximal tubular epithelial cells in vitro: analysis of the effects of interleukin-1 alpha (IL-1 alpha) and other cytokines

• Published in: Nephrology, dialysis, transplantation Vol. 9; no. 6; p. 599
• Main Authors: Boswell, R N, Yard, B A, Schrama, E, van Es, L A, Daha, M R, van der Woude, F J
• Format: Journal Article
• Language: English
• Published: England 1994
• Subjects: Actins - biosynthesis; Actins - drug effects; Actins - genetics; Base Sequence; Blotting, Western; Cells, Cultured; Cycloheximide - pharmacology; DNA, Complementary - biosynthesis; Epithelium - drug effects; Epithelium - metabolism; Humans; Interferon-gamma - pharmacology; Interleukin-1 - pharmacology; Interleukin-2 - pharmacology; Interleukin-6 - biosynthesis; Interleukin-6 - genetics; Kidney Tubules, Proximal - drug effects; Kidney Tubules, Proximal - metabolism; Lipopolysaccharides - pharmacology; Molecular Sequence Data; Polymerase Chain Reaction; Recombinant Proteins; Tumor Necrosis Factor-alpha - pharmacology
• ISSN: 0931-0509
• DOI: 10.1093/ndt/9.6.599

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 alpha), tumour necrosis factor alpha (TNF alpha), interleukin 2 (IL-2), interferon gamma (INF gamma) 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/10(5) PTEC. PTEC cell lines exposed to medium alone produced intermediate amounts of IL-6 with substantial variability between cell lines. Introduction of IL-1 alpha resulted in a dose- and time-dependent increase in IL-6 production by PTEC that was maximal at 1 ng/ml IL-1 alpha at 24 h. All PTEC cell lines showed an increased IL-6 production on exposure to IL-1 alpha varying from 1.3- to 24-fold increase over baseline production. This response was completely blocked by anti-rIL-1 alpha. No significant IL-6 production by PTEC could be induced by TNF alpha, IL-2, IFN gamma, 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 alpha of IL-6 mRNA in a dose-response fashion, indicating that IL-1 alpha regulates IL-6 production at a pretranslational value of protein synthesis.