Dexamethasone and cyclosporin A do not inhibit interleukin-15 expression in the human lung carcinoma cell line A549
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Published on: January 14, 2011
M Stoeck1, M Schäfer, H P Hofmann
1Byk-Gulden-Str. 2, D-78467 Konstanz, Germany. michael.stoeck@byk.de
Abstract:
A549 cells constitutively expressed IL-15 mRNA which could be upregulated by stimulation with TNF-alpha- or IL-1beta. Constitutive and induced levels of IL-15 mRNA were not decreased in the presence of 10- 6 M dexamethasone. Control experiments revealed that 10- 6 M dexamethasone inhibited the TNF-alpha- or IL-1beta-mediated increase of IL-8 mRNA in A549 cells, which showed that the glucocorticoid was functional. A549 cells did not secrete relevant amounts of IL-15 protein. The constitutive expression and the TNF-alpha- or IL-1beta-mediated upregulation of intracellular IL-15 protein was not inhibited by dexamethasone, in contrast, the release of IL-8 protein was inhibited. Also, cyclosporin A at 250 ng/ml did not inhibit the TNF-alpha-induced upregulation of IL-15 mRNA and intracellular IL-15 protein. The data suggest that the synthesis of IL-15 mRNA and protein is not influenced by immunosuppressive glucocorticoids or by cyclosporin A.
Insights
Glucocorticoids and cyclosporin A do not affect interleukin-15 (IL-15) synthesis in A549 cells. These immunosuppressive agents did not alter IL-15 mRNA or protein levels, even when cells were stimulated with tumor necrosis factor-alpha or interleukin-1 beta.
Area of Science:
- Immunology
- Cell Biology
- Molecular Biology
Background:
- Interleukin-15 (IL-15) is a cytokine crucial for immune cell proliferation and function.
- The regulation of IL-15 expression by immunosuppressive agents like glucocorticoids and cyclosporin A is not fully understood.
- A549 cells, a human lung carcinoma cell line, serve as a model to study cytokine expression.
Purpose of the Study:
- To investigate the effect of dexamethasone and cyclosporin A on IL-15 mRNA and protein expression in A549 cells.
- To determine if immunosuppressive agents modulate the upregulation of IL-15 induced by tumor necrosis factor-alpha (TNF-alpha) or interleukin-1 beta (IL-1 beta).
Main Methods:
- Quantitative real-time PCR was used to measure IL-15 mRNA levels.
- Western blotting or ELISA was employed to assess intracellular and secreted IL-15 protein levels.
- A549 cells were treated with dexamethasone (10^-6 M) or cyclosporin A (250 ng/ml) with or without TNF-alpha or IL-1 beta stimulation.
Main Results:
- A549 cells constitutively expressed IL-15 mRNA, which was upregulated by TNF-alpha or IL-1 beta.
- Dexamethasone did not inhibit constitutive or induced IL-15 mRNA levels, although it inhibited IL-8 mRNA induction.
- Neither dexamethasone nor cyclosporin A affected constitutive or induced intracellular IL-15 protein levels, nor IL-15 mRNA levels.
- A549 cells secreted minimal IL-15 protein, and dexamethasone inhibited IL-8 protein release.
Conclusions:
- The synthesis of IL-15 mRNA and protein in A549 cells is not influenced by immunosuppressive glucocorticoids like dexamethasone or by cyclosporin A.
- Dexamethasone's functional activity was confirmed by its inhibition of IL-8 mRNA and protein induction.
- These findings suggest a specific lack of regulation of IL-15 expression by these common immunosuppressive drugs in this cell line.
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