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Christine M. Braun, BA, Shau-Ku Huang, PhD, Gregory G

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1 Corticosteroid modulation of human, antigen-specific Th1 and Th2 responses 
Christine M. Braun, BA, Shau-Ku Huang, PhD, Gregory G. Bashian, BS, Anne Kagey-Sobotka, PhD, Lawrence M. Lichtenstein, MD, PhD, David M. Essayan, MD  Journal of Allergy and Clinical Immunology  Volume 100, Issue 3, Pages (September 1997) DOI: /S (97) Copyright © 1997 Mosby, Inc. Terms and Conditions

2 Fig. 1 Modulation of RW- and TT-stimulated PBMC proliferation by dexamethasone. A, Dose-response data are shown for simultaneous addition of antigen and steroid. Data are presented as percentage of inhibition ± SEM relative to stimulated, steroid-free cultures, subtracted for background with media alone. Background and stimulated mean counts per minute ± SEM were: 1323 ± 109 and 6120 ± 1848 for RW and 1482 ± 166 and 14,003 ± 3229 for TT. N = 9 individual experiments for RW and 7 for TT from six different subjects. B, Efficacy of 10–7 mol/L dexamethasone added at various times after stimulation of PBMC cultures with RW or TT is shown. Data are presented as percentage of inhibition ± SEM relative to stimulated, steroid-free cultures, subtracted for background with media alone at each time point. N = 3 individual experiments at each time point from three different subjects. Journal of Allergy and Clinical Immunology  , DOI: ( /S (97) ) Copyright © 1997 Mosby, Inc. Terms and Conditions

3 Fig. 1 Modulation of RW- and TT-stimulated PBMC proliferation by dexamethasone. A, Dose-response data are shown for simultaneous addition of antigen and steroid. Data are presented as percentage of inhibition ± SEM relative to stimulated, steroid-free cultures, subtracted for background with media alone. Background and stimulated mean counts per minute ± SEM were: 1323 ± 109 and 6120 ± 1848 for RW and 1482 ± 166 and 14,003 ± 3229 for TT. N = 9 individual experiments for RW and 7 for TT from six different subjects. B, Efficacy of 10–7 mol/L dexamethasone added at various times after stimulation of PBMC cultures with RW or TT is shown. Data are presented as percentage of inhibition ± SEM relative to stimulated, steroid-free cultures, subtracted for background with media alone at each time point. N = 3 individual experiments at each time point from three different subjects. Journal of Allergy and Clinical Immunology  , DOI: ( /S (97) ) Copyright © 1997 Mosby, Inc. Terms and Conditions

4 Fig. 2 Modulation of antigen-driven Th1 and Th2 proliferation by corticosteroids. RW-specific T-cell clones were stimulated in the presence of various concentrations of each of three corticosteroids: dexamethasone, budesonide, and hydrocortisone. Data are presented as percentage of inhibition ± SEM relative to stimulated, steroid-free cultures, subtracted for background with media alone. Background and stimulated mean counts per minute ± SEM for Th1 clones were: 4450 ± 1659 and 80,145 ± 34,522 in dexamethasone assays; 2785 ± 1449 and 77,933 ± 26,367 in budesonide assays; and 1449 ± 652 and 58,516 ± 29,298 in hydrocortisone assays. Background and stimulated mean counts per minute ± SEM for Th2 clones were: 2523 ± 1913 and 45,201 ± 14,653 in dexamethasone assays; 3918 ± 2104 and 83,232 ± 28,915 in budesonide assays; and 2873 ± 1817 and 37,065 ± 10,077 in hydrocortisone assays. N = 5 individual experiments with Th1 clones and 4 individual experiments with Th2 clones. Journal of Allergy and Clinical Immunology  , DOI: ( /S (97) ) Copyright © 1997 Mosby, Inc. Terms and Conditions

5 Fig. 2 Modulation of antigen-driven Th1 and Th2 proliferation by corticosteroids. RW-specific T-cell clones were stimulated in the presence of various concentrations of each of three corticosteroids: dexamethasone, budesonide, and hydrocortisone. Data are presented as percentage of inhibition ± SEM relative to stimulated, steroid-free cultures, subtracted for background with media alone. Background and stimulated mean counts per minute ± SEM for Th1 clones were: 4450 ± 1659 and 80,145 ± 34,522 in dexamethasone assays; 2785 ± 1449 and 77,933 ± 26,367 in budesonide assays; and 1449 ± 652 and 58,516 ± 29,298 in hydrocortisone assays. Background and stimulated mean counts per minute ± SEM for Th2 clones were: 2523 ± 1913 and 45,201 ± 14,653 in dexamethasone assays; 3918 ± 2104 and 83,232 ± 28,915 in budesonide assays; and 2873 ± 1817 and 37,065 ± 10,077 in hydrocortisone assays. N = 5 individual experiments with Th1 clones and 4 individual experiments with Th2 clones. Journal of Allergy and Clinical Immunology  , DOI: ( /S (97) ) Copyright © 1997 Mosby, Inc. Terms and Conditions

6 Fig. 3 Modulation of cytokine gene expression by dexamethasone in RW- and TT-stimulated PBMCs. Representative RT-PCR amplification products for IL-4, IL-5, IFN-γ, and IL-13 are shown for each of seven different PBMC culture conditions from a single experiment. Adequate normalization of RNA was confirmed by equivalent β-actin gene expression at subsaturating cycle number (row 1). The DNA size marker is shown in lane 8. N = 3 individual experiments in three different subjects. Dex, Dexamethasone. Journal of Allergy and Clinical Immunology  , DOI: ( /S (97) ) Copyright © 1997 Mosby, Inc. Terms and Conditions

7 Fig. 4 Modulation of cytokine gene expression in RW-stimulated Th1 (A) and Th2 (B) clones by corticosteroids. Representative RT-PCR amplification products for IFN-γ and IL-13 in Th1 clones and IL-4, IL-5, and IL-13 in Th2 clones are shown for each of seven different culture conditions from a single experiment. Adequate normalization of RNA was confirmed by equivalent β-actin gene expression at subsaturating cycle number (row 1). The DNA size marker is shown in lane 8. N = 3 individual experiments with Th1 clones and 3 individual experiments with Th2 clones. Dex, Dexamethasone; Bud, budesonide; HC, hydrocortisone. Journal of Allergy and Clinical Immunology  , DOI: ( /S (97) ) Copyright © 1997 Mosby, Inc. Terms and Conditions

8 Fig. 4 Modulation of cytokine gene expression in RW-stimulated Th1 (A) and Th2 (B) clones by corticosteroids. Representative RT-PCR amplification products for IFN-γ and IL-13 in Th1 clones and IL-4, IL-5, and IL-13 in Th2 clones are shown for each of seven different culture conditions from a single experiment. Adequate normalization of RNA was confirmed by equivalent β-actin gene expression at subsaturating cycle number (row 1). The DNA size marker is shown in lane 8. N = 3 individual experiments with Th1 clones and 3 individual experiments with Th2 clones. Dex, Dexamethasone; Bud, budesonide; HC, hydrocortisone. Journal of Allergy and Clinical Immunology  , DOI: ( /S (97) ) Copyright © 1997 Mosby, Inc. Terms and Conditions

9 Fig. 5 Dose-dependent modulation of cytokine production by corticosteroids in Th1 and Th2 clones. Secretion of IFN-γ by Th1 clones (A) and IL-4 by Th2 clones (B) in the presence of each of three corticosteroids is shown. For dexamethasone and budesonide, the high dose was 10–7 mol/L, the medium dose was 10–9 mol/L, and the low dose was 10–10 mol/L. For hydrocortisone, the high dose was 10–6 mol/L, the medium dose was 10–8 mol/L, and the low dose was 10–9 mol/L. Data are presented as percentage of inhibition ± SEM relative to stimulated, steroid-free control cultures. Mean IFN-γ production from stimulated Th1 controls = 6147 ± 3084 pg/ml; mean IL-4 production from stimulated Th2 controls = 872 ± 193 pg/ml. Dex, Dexamethasone; Bud, budesonide; HC, hydrocortisone. Journal of Allergy and Clinical Immunology  , DOI: ( /S (97) ) Copyright © 1997 Mosby, Inc. Terms and Conditions

10 Fig. 5 Dose-dependent modulation of cytokine production by corticosteroids in Th1 and Th2 clones. Secretion of IFN-γ by Th1 clones (A) and IL-4 by Th2 clones (B) in the presence of each of three corticosteroids is shown. For dexamethasone and budesonide, the high dose was 10–7 mol/L, the medium dose was 10–9 mol/L, and the low dose was 10–10 mol/L. For hydrocortisone, the high dose was 10–6 mol/L, the medium dose was 10–8 mol/L, and the low dose was 10–9 mol/L. Data are presented as percentage of inhibition ± SEM relative to stimulated, steroid-free control cultures. Mean IFN-γ production from stimulated Th1 controls = 6147 ± 3084 pg/ml; mean IL-4 production from stimulated Th2 controls = 872 ± 193 pg/ml. Dex, Dexamethasone; Bud, budesonide; HC, hydrocortisone. Journal of Allergy and Clinical Immunology  , DOI: ( /S (97) ) Copyright © 1997 Mosby, Inc. Terms and Conditions

11 Fig. 6 Dose-dependent modulation of IL-13 production by corticosteroids in Th1 and Th2 clones. Secretion of IL-13 by Th1 (A) and Th2 (B) clones in the presence of each of three corticosteroids is shown for the same culture conditions indicated in Fig. 5. Data are presented as percentage of inhibition ± SEM relative to stimulated, steroid-free cultures. Mean IL-13 production from stimulated controls = 688 ± 102 pg/ml for Th1 clones and 3531 ± 1,119 pg/ml for Th2 clones. Dex, Dexamethasone; Bud, budesonide; HC, hydrocortisone. Journal of Allergy and Clinical Immunology  , DOI: ( /S (97) ) Copyright © 1997 Mosby, Inc. Terms and Conditions

12 Fig. 6 Dose-dependent modulation of IL-13 production by corticosteroids in Th1 and Th2 clones. Secretion of IL-13 by Th1 (A) and Th2 (B) clones in the presence of each of three corticosteroids is shown for the same culture conditions indicated in Fig. 5. Data are presented as percentage of inhibition ± SEM relative to stimulated, steroid-free cultures. Mean IL-13 production from stimulated controls = 688 ± 102 pg/ml for Th1 clones and 3531 ± 1,119 pg/ml for Th2 clones. Dex, Dexamethasone; Bud, budesonide; HC, hydrocortisone. Journal of Allergy and Clinical Immunology  , DOI: ( /S (97) ) Copyright © 1997 Mosby, Inc. Terms and Conditions

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