Downregulation of basophil-derived IL-4 and in vivo TH2 IgE responses by serotonin and other organic cation transporter 3 ligands Elke Schneider, PhD,

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Presentation transcript:

Downregulation of basophil-derived IL-4 and in vivo TH2 IgE responses by serotonin and other organic cation transporter 3 ligands Elke Schneider, PhD, François Machavoine, MSc, Rachel Bricard-Rignault, MSc, Mélanie Levasseur, MSc, Anne France Petit-Bertron, PhD, Sophie Gautron, PhD, Jean-Antoine Ribeil, MD, PhD, Jean-Marie Launay, MD, PhD, Salah Mecheri, PhD, Francine Côté, PhD, Michel Dy, PhD Journal of Allergy and Clinical Immunology Volume 128, Issue 4, Pages 864-871.e2 (October 2011) DOI: 10.1016/j.jaci.2011.04.043 Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 Basophils are proficient IL-4 producers in response to several stimuli. A, Sorting of basophils isolated ex vivo from BMCs or derived from culture. B, IL-4 production by bone marrow populations containing different proportions of basophils. IL-4 was measured in supernatants after a 24-hour incubation in the presence of the stimuli indicated (n = 4). Ca iono, Calcium ionophore. Journal of Allergy and Clinical Immunology 2011 128, 864-871.e2DOI: (10.1016/j.jaci.2011.04.043) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 5-HT downregulates IL-4 production by basophils on protein and transcription levels. A, Dose-response curve (n = 4). HA, Histamine. B, 5-HT, histamine, and ciproxifan (CPF; all at 5 × 10−4 mol/L) inhibit IL-3–induced IL-4 production by basophils sorted ex vivo (n = 3). ∗P ≤ .05. ∗∗P ≤ .001. C, 5-HT does not impair basophil viability. 7-AAD, 7-Aminoactinomycin D. D, 5-HT inhibits IL4 mRNA expression. E, 5-HT inhibits IL-4 production in response to several stimuli (n = 3). Ca iono, Calcium ionophore. Journal of Allergy and Clinical Immunology 2011 128, 864-871.e2DOI: (10.1016/j.jaci.2011.04.043) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 3 SERT mediates the high-affinity uptake of 5-HT by murine basophils but not its inhibitory effect on IL-4 production. A, Concomitant OCT3 and SERT expression in purified bone marrow–derived CD49b+ckit− basophils. β2m, β2-Microglobulin. B, Kinetics of 5-HT uptake (n = 4). C, Displacement of tritiated histamine taken up by BMCs by unlabeled monoamines (n = 3). HA, Histamine. D, 5-HT–induced inhibition of basophil activities is abolished in Oct3−/− mice (n = 3). WT, Wild-type. Journal of Allergy and Clinical Immunology 2011 128, 864-871.e2DOI: (10.1016/j.jaci.2011.04.043) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 4 OCT3 ligands decrease basophil functions in vivo. A, BMCs from mice receiving daily injections of IL-33 for 4 days with or without coinjected histamine (HA), serotonin (5-HT), or ciproxifan (CPF) were recovered on day 4 and restimulated with IL-33 (10 ng/mL) for 24 hours compared with saline controls (n = 3). B, IgE levels were measured after immunization with papain in sera of mice treated or not with OCT3 ligands (n = 3). CB, Clobenpropit. ∗P ≤ .01; ∗∗P ≤ .002. Journal of Allergy and Clinical Immunology 2011 128, 864-871.e2DOI: (10.1016/j.jaci.2011.04.043) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E1 5-HT inhibits IL-3–induced IL-6 and histamine production. Basophil-enriched NK1.1−CD49b+ cells (2.5 × 105/mL) were incubated for 24 hours in the presence of IL-3 (10 ng/mL) with or without 5-HT (5 × 10−4 mol/L). Data are presented as means ± SEMs from 4 separate experiments. CTRL, Control. Journal of Allergy and Clinical Immunology 2011 128, 864-871.e2DOI: (10.1016/j.jaci.2011.04.043) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E2 The CYP450 ligand ketoconazole (KTO) inhibits IL-3–induced IL-4 production by basophils and competes with 5-HT (n = 3). Journal of Allergy and Clinical Immunology 2011 128, 864-871.e2DOI: (10.1016/j.jaci.2011.04.043) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions