Regulation of 15-lipoxygenase expression in lung epithelial cells by interleukin-4

Brinckmann, Roland; Topp, Max S.; Zalán, Ildikó; Heydeck, Dagmar; Ludwig, P.; Kühn, Hartmut; Berdel, Wolfgang E.; Habenicht, Andreas J. R.

doi:10.1042/bj3180305

Cited by 90 publications

(76 citation statements)

References 53 publications

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“…11 These effects were inhibited by IFN-␥, as was seen in IL-4-induced 15-LO-1 expression. 10 Similar results have been observed by Brinkmann et al 12 in a human epithelial cell line, A549 in which induction of 15-LO-1 by IL-13 and IL-4 was demonstrated.…”

Section: Introductionsupporting

confidence: 77%

Ku autoantigen (DNA helicase) is required for interleukins-13/-4-induction of 15-Lipoxygenase-1 gene expression in human epithelial cells

2000

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As reported previously in human monocytes, a human lung epithelial cell line, A549, showed de novo induction of 15-Lipoxygenase-1 (15-LO-1) in response to interleukins-13 (IL-13) and −4 (IL-4). In this cell line, 15-LO-1 expression, by RT-PCR and western blotting, was observed following 6 and 24 h of exposure to human IL-13 (ED 50 5 ng/ml) and IL-4 (ED 50 0.2 ng/ml). We have previously shown that no cis-acting regulatory elements exist within the 15-LO-1 promoter region. To define IL-13 and IL-4 responsive trans-acting elements, we identified a region (DP2: −353 to −304 bp site) within the 15-LO-1 promoter (by footprinting experiments) to which IL-13-responsive elements (or factors) bind specifically (Kelavkar et al, 1998, Mol Biol Rep 25, 173-182). To further delineate this region, we constructed (by site-directed mutagenesis) several deletion mutants in the 'LOPB5' region containing the 29 bp within the −353 to −304 bp of the DP2 core element. These were: DP3 (site totally deleted), DP4 (5 bp deleted at the center of the site), DP5 (8 bp at the 5Ј-end of the site) and DP6 (13 bp at the 3Ј-end of the site). Cotransfection of these deletion constructs (driving luciferase reporter genes) was associated with 90% (DP4, DP5 and DP6) or 100% (DP3) abrogation of promoter activity at 24 h. Purification of nuclear protein extracts from IL-13 and IL-4-stimulated A549 cells, using a DP2 core containing affinity column, identified a 150 kDa protein under non-denaturing conditions, and two, 70 and 85 kDa proteins under denaturing conditions. These were not detectable by Coomassie blue staining in control nuclear protein extracts. Matrix assisted laser desorption ionization mass spectrometry (MALDI-MS) of the tryptic digests of these proteins, identified one as the 86 kDA Lupus KU autoantigen protein P86 and the second as the 70 kDa Lupus KU autoantigen protein P70. Gel shift and supershift experiments using monoclonal antibodies toward Ku antigen and its individual subunits, and utilizing DP2 and other mutant oligonucleotides with purified nuclear protein extracts from control and cytokine-treated A549 cells, confirmed our findings. Furthermore, electroporation of neutralizing anti-Ku70, Ku 80 and Ku70/80 antibodies into A549 cells totally suppressed IL-13 and IL-4-stimulated 15-LO-1 induction in these cells. Further, immunoprecipitation experiments data suggests that IL-4 and IL-13 activate Ku antigens and 15-LO-1 expression through distinct signaling events. In summary, in A549 cells, Ku antigen is induced in response to the cytokines, IL-13 and −4, and a 29 bp region within the −353 to −304 bp region of the 15-LO-1 promoter is required for its binding and subsequent induction of 15-LO-1 gene expression. The findings may provide an important link between the established dysregulated function of Ku antigen in auto-immune diseases, such as systemic lupus erythematosus and thyroiditis, and the increasingly recognized 'anti-inflammatory' role of 15-LO-1. Genes and Immunity (2000) 1, 237-250.

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Section: Introductionsupporting

confidence: 77%

Ku autoantigen (DNA helicase) is required for interleukins-13/-4-induction of 15-Lipoxygenase-1 gene expression in human epithelial cells

2000

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“…These include platelet-type 12-LO, leukocyte-type 12-LO, epidermaltype 12-LO, and 12-(R)-LO (36). In addition, 15-lipoxygenase 1 is capable of synthesizing 12-HpETE as a minor product and has been shown to be expressed in A549 cells in response to IL-4 and IL-13 (38). Alternatively, bacterial infection could simply stimulate the production of increased amounts of HXA 3 precursors such as arachidonic acid, thus providing more substrate for a constitutively active 12-LO enzyme.…”

Section: Discussionmentioning

confidence: 99%

Polymorphonuclear Cell Transmigration Induced by Pseudomonas aeruginosa Requires the Eicosanoid Hepoxilin A3

Hurley

Siccardi

Mrsny

et al. 2004

The Journal of Immunology

161

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Lung inflammation resulting from bacterial infection of the respiratory mucosal surface in diseases such as cystic fibrosis and pneumonia contributes significantly to the pathology. A major consequence of the inflammatory response is the recruitment and accumulation of polymorphonuclear cells (PMNs) at the infection site. It is currently unclear what bacterial factors trigger this response and exactly how PMNs are directed across the epithelial barrier to the airway lumen. An in vitro model consisting of human PMNs and alveolar epithelial cells (A549) grown on inverted Transwell filters was used to determine whether bacteria are capable of inducing PMN migration across these epithelial barriers. A variety of lung pathogenic bacteria, including Klebsiella pneumoniae, Escherichia coli, and Pseudomonas aeruginosa are indeed capable of inducing PMN migration across A549 monolayers. This phenomenon is not mediated by LPS, but requires live bacteria infecting the apical surface. Bacterial interaction with the apical surface of A549 monolayers results in activation of epithelial responses, including the phosphorylation of ERK1/2 and secretion of the PMN chemokine IL-8. However, secretion of IL-8 in response to bacterial infection is neither necessary nor sufficient to mediate PMN transepithelial migration. Instead, PMN transepithelial migration is mediated by the eicosanoid hepoxilin A3, which is a PMN chemoattractant secreted by A549 cells in response to bacterial infection in a protein kinase C-dependent manner. These data suggest that bacterial-induced hepoxilin A3 secretion may represent a previously unrecognized inflammatory mechanism occurring within the lung epithelium during bacterial infections.

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“…The amino acid sequence similarity between these enzymes is only 40% (3). IL-4 and/or IL-13 induce the expression of 15-LO-1 in cultured mast cells, monocytes, and epithelial cells (4,8,9). Thus, these cytokines may act as important physiological/pathophysiological regulators of 15-LO-1 expression in vivo.…”

mentioning

confidence: 99%

Eoxins are proinflammatory arachidonic acid metabolites produced via the 15-lipoxygenase-1 pathway in human eosinophils and mast cells

Feltenmark¹,

Gautam²,

Brunnström³

et al. 2008

Proc. Natl. Acad. Sci. U.S.A.

148

143

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Human eosinophils contain abundant amounts of 15-lipoxygenase (LO)-1. The biological role of 15-LO-1 in humans, however, is unclear. Incubation of eosinophils with arachidonic acid led to formation of a product with a UV absorbance maximum at 282 nm and shorter retention time than leukotriene (LT)C4 in reverse-phase HPLC. Analysis with positive-ion electrospray tandem MS identified this eosinophil metabolite as 14,15-LTC4. This metabolite could be metabolized to 14,15-LTD4 and 14,15-LTE4 in eosinophils. Because eosinophils are such an abundant source of these metabolites and to avoid confusion with 5-LO-derived LTs, we suggest the names eoxin (EX)C4, -D4, and -E4 instead of 14,15-LTC4, -D4, and -E4, respectively. Cord blood-derived mast cells and surgically removed nasal polyps from allergic subjects also produced EXC4. Incubation of eosinophils with arachidonic acid favored the production of EXC4, whereas challenge with calcium ionophore led to exclusive formation of LTC4. Eosinophils produced EXC4 after challenge with the proinflammatory agents LTC4, prostaglandin D2, and IL-5, demonstrating that EXC4 can be synthesized from the endogenous pool of arachidonic acid. EXs induced increased permeability of endothelial cell monolayer in vitro, indicating that EXs can modulate and enhance vascular permeability, a hallmark of inflammation. In this model system, EXs were 100 times more potent than histamine and almost as potent as LTC4 and LTD4. Taken together, this article describes the formation of proinflammatory EXs, in particular in human eosinophils but also in human mast cells and nasal polyps.

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Regulation of 15-lipoxygenase expression in lung epithelial cells by interleukin-4

Cited by 90 publications

References 53 publications

Ku autoantigen (DNA helicase) is required for interleukins-13/-4-induction of 15-Lipoxygenase-1 gene expression in human epithelial cells

Ku autoantigen (DNA helicase) is required for interleukins-13/-4-induction of 15-Lipoxygenase-1 gene expression in human epithelial cells

Polymorphonuclear Cell Transmigration Induced by Pseudomonas aeruginosa Requires the Eicosanoid Hepoxilin A3

Eoxins are proinflammatory arachidonic acid metabolites produced via the 15-lipoxygenase-1 pathway in human eosinophils and mast cells

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