Dexamethasone suppresses gene expression and production of IL-13 by human mast cell line and lung mast cells Toshiaki Fushimi, MD, Hiroshi Okayama, MD,

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Dexamethasone suppresses gene expression and production of IL-13 by human mast cell line and lung mast cells Toshiaki Fushimi, MD, Hiroshi Okayama, MD, Sanae Shimura, MD, Hiroki Saitoh, MD, Kunio Shirato, MD Journal of Allergy and Clinical Immunology Volume 102, Issue 1, Pages 134-142 (July 1998) DOI: 10.1016/S0091-6749(98)70064-8 Copyright © 1998 Mosby, Inc. Terms and Conditions

Fig. 1 Effect of number of PCR cycles on amplification of IL-13 cDNA. Different amounts of same cDNA template were amplified for 15 to 32 cycles with IL-13–specific primers. There was a linear relationship between level of amplified IL-13 cDNA fragment and amount of template at 15 to 26 cycles, but not at more than 29 cycles. Journal of Allergy and Clinical Immunology 1998 102, 134-142DOI: (10.1016/S0091-6749(98)70064-8) Copyright © 1998 Mosby, Inc. Terms and Conditions

Fig. 2 Time course of IL-13 cDNA in HMC-1 cells stimulated with PMA and ionomycin. A, Southern blot analysis of PMA/ionomycin-stimulated HMC-1 cells. PMA/ionomycin caused rapid increase in IL-13 cDNA, which peaked at 2 hours and then decreased rapidly thereafter. B, Time course of IL-13 cDNA increase determined by image analyzer. Radioactivity of IL-13 cDNA band is normalized by that of β-actin cDNA band and is shown as percent of each PMA/ionomycin-stimulated value. Peak was observed between 1 and 3 hours, and it decreased thereafter. Journal of Allergy and Clinical Immunology 1998 102, 134-142DOI: (10.1016/S0091-6749(98)70064-8) Copyright © 1998 Mosby, Inc. Terms and Conditions

Fig. 3 The effect of dexamethasone on the PMA/ionomycin-stimulated expression of IL-13 gene in HMC-1 cells. A, Southern blot analysis. Length of amplified products was 501 bp and 1017 bp for IL-13 cDNA and β-actin cDNA, respectively. B, Effect of dexamethasone on PMA/ionomycin-stimulated expression of IL-13 gene determined by image analyzer. PMA/ionomycin significantly increased level of IL-13 cDNA, and dexamethasone significantly suppressed it by 68% at 10–6 mol/L. Radioactivity of IL-13 cDNA band is normalized by that of β-actin cDNA band and is shown as percent of each PMA/ionomycin-stimulated value. Each point represents mean ± SEM of four experiments. ***p < 0.001 compared with PMA/ionomycin-stimulated value. Journal of Allergy and Clinical Immunology 1998 102, 134-142DOI: (10.1016/S0091-6749(98)70064-8) Copyright © 1998 Mosby, Inc. Terms and Conditions

Fig. 4 Dose-response of dexamethasone on PMA/ionomycin-stimulated expression of IL-13 gene in HMC-1 cells. Radioactivity of IL-13 cDNA band is normalized by that of β-actin cDNA band and is shown as percent of each PMA/ionomycin-stimulated value. Level of IL-13 cDNA was suppressed with increasing concentrations of dexamethasone. Journal of Allergy and Clinical Immunology 1998 102, 134-142DOI: (10.1016/S0091-6749(98)70064-8) Copyright © 1998 Mosby, Inc. Terms and Conditions

Fig. 5 A, Effect of dexamethasone on PMA/ionomycin-stimulated expression of IL-13 gene in freshly isolated human lung mast cells. IL-13 cDNA band was observed without stimulation. When stimulated with 10 ng/ml PMA and 1 μmol/L ionomycin, thick band was identified. Intensity of band was decreased with 10–6 mol/L dexamethasone. B, Effect of dexamethasone on IL-13 production in PMA/ionomycin-stimulated human lung mast cells. Unstimulated human lung mast cells produced low level of IL-13. With PHA/ionomycin–stimulation, level of IL-13 was increased, and level was suppressed by dexamethasone. Mean value of two experiments is shown. Journal of Allergy and Clinical Immunology 1998 102, 134-142DOI: (10.1016/S0091-6749(98)70064-8) Copyright © 1998 Mosby, Inc. Terms and Conditions