Delivery of the interleukin-1 type ii receptor cdna inhibits interleurin-8 production in human fibroblasts

Daren L Knoell, C. B. Marsh, M. D. Wewers

Research output: Contribution to journalArticle

Abstract

Communication between monocytes, fibroblasts, and hepatocytes in the liver constitute an important feedback pathway in inflammation. Interleukin-1 (ILI) is a key mediator in all three cell types and is directly involved in the liver response to inflammatory stimuli. Recent evidence suggests that the Interleukin-1 Type II receptor (IL-1 Ilr) serves as an endogenous inhibitor of IL-1. We hypothesized that overexpression of the soluble IL-1 Ilr in hepatocytes would inhibit IL-1β mediated inflammation by hepatocytes, fibroblasts, and monocytes. To test this hypothesis, transfection of the human hepatoma cell line HepG2 was optimized using a β-galactosidase reporter plasmid and Lipofectamine®, A mammalian expression vector containing the human IL-1 Ilr cDNA (provided by U.Gubler) was then used to transfect HepG2 cells. Soluble IL-1 Ilr was detectable by ELISA in the supernatents of transfected HepG2 cells (1500-4500 pg/ml) 3-5 days after transfection. The same supernatents were found to bind IL-1 as determined by decreased IL-I ïïr ELISA detection in the presence of recombinant IL-1β. When transfected HepG2 supernatents were incubated with human gingival fibroblasts in the presence of IL-1β (100 pg/ml), interleukin-8 production was completely inhibited (as measured by ELISA in supernatents) whereas, IL-8 induction was unaffected in fibroblasts treated with control and sham-transfected conditioned médias (ANOVA p=.001). IL-1 induction of IL-8 in HepG2 cells and human monocytes was not inhibited by the presence of soluble IL-1 Ilr from transfected cells. These results suggest that the inhibitory effects of soluble IL-1 Ilr are cell dependent and demonstrate the potential for gene delivery to parenchymal cells of the liver to inhibit cytokine mediated inflammation.

Original languageEnglish (US)
JournalFASEB Journal
Volume10
Issue number6
StatePublished - Dec 1 1996

Fingerprint

Interleukin-1 Receptors
interleukin-1
Fibroblasts
Interleukin-1
fibroblasts
receptors
hepatocytes
Hep G2 Cells
interleukin-8
Interleukin-8
Liver
monocytes
Monocytes
Hepatocytes
Enzyme-Linked Immunosorbent Assay
inflammation
Inflammation
enzyme-linked immunosorbent assay
cells
transfection

ASJC Scopus subject areas

  • Agricultural and Biological Sciences (miscellaneous)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Cell Biology

Cite this

Delivery of the interleukin-1 type ii receptor cdna inhibits interleurin-8 production in human fibroblasts. / Knoell, Daren L; Marsh, C. B.; Wewers, M. D.

In: FASEB Journal, Vol. 10, No. 6, 01.12.1996.

Research output: Contribution to journalArticle

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