Interleukin-4 induces 15-lipoxygenase-1 expression in human orbital fibroblasts from patients with Graves disease. Evidence for anatomic site-selective actions of Th2 cytokines

J Biol Chem. 2006 Jul 7;281(27):18296-306. doi: 10.1074/jbc.M603484200. Epub 2006 May 4.

Abstract

Orbital fibroblasts orchestrate tissue remodeling in Graves disease, at least in part, because they exhibit exaggerated responses to proinflammatory cytokines. A hallmark of late stage orbital disease is vision-threatening fibrosis, the molecular basis of which remains uncertain. We report here that the Th2 cytokines, interleukin (IL)-4 and IL-13, can induce in these cells the expression of 15-lipoxygenase-1 (15-LOX-1) and in so doing up-regulate the production of 15-hydroxyeicosatetraenoic acid. IL-4 increases 15-LOX-1 protein levels through pretranslational actions. The increased steady-state 15-LOX-1 mRNA is independent of ongoing protein synthesis and involves very modestly increased gene promoter activity. Importantly, IL-4 substantially enhances 15-LOX-1 transcript stability, activity that localizes to a 293-bp sequence of the 3'-untranslated region. IL-4 activates Jak2 in orbital fibroblasts. Interrupting signaling through that pathway, either with the specific chemical inhibitor, AG490, or by transiently transfecting the cells with a Jak2 dominant negative mutant kinase, attenuates the 15-LOX-1 induction. Interferongamma, a Th1 cytokine, could block this induction by attenuating IL-4-dependent mRNA stabilization. 15-LOX-1 protein and its mRNA were undetectable in IL-4-treated dermal fibroblasts, despite comparable levels of cell surface IL-4 receptor and phosphorylated Jak2 and STAT6. Our findings suggest that orbital connective tissues may represent a site of localized 15-hydroxyeicosatetraenoic acid generation resulting from cell type-specific 15-LOX-1 mRNA stabilization by IL-4. These results may have relevance to the pathogenesis of orbital Graves disease, an inflammatory autoimmune condition that gives way to extensive fibrosis associated with a Th2 response.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arachidonate 15-Lipoxygenase / biosynthesis*
  • Arachidonate 15-Lipoxygenase / genetics
  • Autoimmunity
  • Dexamethasone / pharmacology
  • Enzyme Induction / drug effects
  • Fibroblasts / enzymology*
  • Fibroblasts / immunology
  • Fibroblasts / pathology
  • Gene Expression Regulation, Enzymologic / drug effects*
  • Graves Disease / enzymology*
  • Graves Disease / immunology
  • Graves Disease / pathology
  • Humans
  • Interferon-gamma / pharmacology
  • Interleukin-13 / immunology
  • Interleukin-13 / pharmacology
  • Interleukin-4 / immunology
  • Interleukin-4 / pharmacology*
  • Orbit / pathology
  • Organ Specificity / immunology
  • Th2 Cells / immunology

Substances

  • Interleukin-13
  • Interleukin-4
  • Dexamethasone
  • Interferon-gamma
  • Arachidonate 15-Lipoxygenase