Early suppression of basophil activation during allergen-specific immunotherapy by histamine receptor 2  Natalija Novak, MD, Nihal Mete, MD, Caroline.

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Early suppression of basophil activation during allergen-specific immunotherapy by histamine receptor 2  Natalija Novak, MD, Nihal Mete, MD, Caroline Bussmann, MD, Laura Maintz, MD, Thomas Bieber, MD, PhD, Mübeccel Akdis, MD, PhD, Judith Zumkehr, MSc, Marek Jutel, MD, Cezmi Akdis, MD  Journal of Allergy and Clinical Immunology  Volume 130, Issue 5, Pages 1153-1158.e2 (November 2012) DOI: 10.1016/j.jaci.2012.04.039 Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 Basophil H2R/H4R mRNA expression is significantly upregulated during the first 6 hours of ultrarush immunotherapy. H2R (A) and H4R (B) mRNA expression levels of purified basophils are shown normalized to mRNA expression of 18s as a housekeeping gene on day 0 before VIT, 6 hours after the beginning of VIT, and on days 2 and 4 (n = 7). *P < .05. Journal of Allergy and Clinical Immunology 2012 130, 1153-1158.e2DOI: (10.1016/j.jaci.2012.04.039) Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 Absolute number of viable basophils in peripheral blood decreases during the build-up phase of ultrarush immunotherapy. A and B, The absolute number of basophils (Fig 2, A) and eosinophils (Fig 2, B) in the peripheral blood (differential blood count) of 13 patients (mean ± SEM) is depicted. C, Percentages of nonapoptotic, nonnecrotic, viable basophils in the peripheral blood of 7 patients during the build-up phase of ultrarush VIT. Values are presented as means ± SEMs. *P < .05 and **P < .01. Journal of Allergy and Clinical Immunology 2012 130, 1153-1158.e2DOI: (10.1016/j.jaci.2012.04.039) Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 3 H2R suppresses IgE/FcεRI cross-linking–induced basophil activation. A, The mean ± SEM value of the percentage of suppression of basophil activation normalized to the CD203c surface expression of FcεRI-activated control cells and cells after preincubation with increasing amounts of dimaprit or forskolin for 6 experiments is depicted on the y-axis, whereas concentrations of dimaprit/forskolin are shown on the x-axis. B, Percentages of CD63 expression of CD123+/HLA-DR− basophils from whole blood samples with and without FcεRI-cross linking by anti-IgE mAb after preincubation with 10−4 mol/L dimaprit (dima), 10−4 mol/L forskolin (forsk), or both are depicted. C, Mean ± SD values of the percentage of CD63-expressing basophils of 10 independent experiments are shown. *P < .05 and **P < .01. Journal of Allergy and Clinical Immunology 2012 130, 1153-1158.e2DOI: (10.1016/j.jaci.2012.04.039) Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 4 Basophil IL-4 and IL-8 release and sulfidoleukotriene release of basophils decreases after preincubation with dimaprit (dima) or forskolin. A and B, Mean ± SEM values of IL-4 (Fig 4, A) and IL-8 (Fig 4, B) levels detected by using Flex Set assays after 12 hours of 12 experiments (n = 6 experiments for preincubation with forskolin) are shown. C, A dose-dependent decrease in sulfidoleukotriene release depicted as mean ± SEM values on the y-axis for 6 experiments is shown. D, Mean ± SEM values of histamine released from basophils after 20 minutes of FcεRI stimulation with or without dimaprit preincubation for 6 experiments are depicted on the y-axis. *P < .05. **P < .01. Journal of Allergy and Clinical Immunology 2012 130, 1153-1158.e2DOI: (10.1016/j.jaci.2012.04.039) Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E1 PBMC H2R mRNA expression is significantly upregulated during the first 5 days of ultrarush immunotherapy. H1R (A), H2R (B), and H4R (C) mRNA expression levels (mean ± SEM values of 12 experiments for PBMCs) are illustrated normalized to expression of 18s as a housekeeping gene. *P < .05. Journal of Allergy and Clinical Immunology 2012 130, 1153-1158.e2DOI: (10.1016/j.jaci.2012.04.039) Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions