Combination treatment with omalizumab and rush immunotherapy for ragweed-induced allergic rhinitis: Inhibition of IgE-facilitated allergen binding Sven.

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Combination treatment with omalizumab and rush immunotherapy for ragweed-induced allergic rhinitis: Inhibition of IgE-facilitated allergen binding Sven Klunker, PhD, Lavina R. Saggar, MSc, Vicki Seyfert-Margolis, PhD, Adam L. Asare, PhD, Thomas B. Casale, MD, Stephen R. Durham, MD, James N. Francis, PhD Journal of Allergy and Clinical Immunology Volume 120, Issue 3, Pages 688-695 (September 2007) DOI: 10.1016/j.jaci.2007.05.034 Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 Schematic of the IgE-FAB assay. Journal of Allergy and Clinical Immunology 2007 120, 688-695DOI: (10.1016/j.jaci.2007.05.034) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 Characterization of the FAB assay. A, Ragweed-specific IgE–containing serum (n = 3) and variable concentrations of ragweed allergen were preincubated for 1 hour at 37°C before addition to EBV-transformed B cells for 1 hour at 4°C. B, High ragweed-specific IgE–containing serum (n = 3) was serially diluted with assay buffer and incubated with 1 μg/mL allergen, as above. C, Indicator serum (n = 3) from patients with high ragweed-specific IgE was heat inactivated at 56°C for 30 minutes, and binding to B cells was compared with that of untreated serum in the assay, as above. D, B cells were pretreated with an anti-CD23 blocking antibody or isotype control antibody for 30 minutes at 4°C and used in the assay, as above. E, Equal volumes of immunotherapy serum were added to indicator serum and various allergen concentrations in the FAB assay. All data are shown as mean (± SE) percentages of B cells bound to allergen-IgE complexes. RW, Ragweed; IT, immunotherapy. Journal of Allergy and Clinical Immunology 2007 120, 688-695DOI: (10.1016/j.jaci.2007.05.034) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 3 Assessment of serum inhibitory properties during the study. Inhibitory activity of serum was tested in the FAB assay, where binding of indicator serum alone was normalized to 100%. Data are shown as means ± SE. Differences between baseline measurements and other time points were assessed by using the Friedman nonparametric repeated-measures test (∗P < .05). Differences between placebo and immunotherapy groups were assessed by using a Mann-Whitney U test (†P < .05). IT, Immunotherapy; OM, omalizumab. Journal of Allergy and Clinical Immunology 2007 120, 688-695DOI: (10.1016/j.jaci.2007.05.034) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 4 Comparison of inhibitory responses with either crude ragweed allergen extract or the major ragweed allergen Amb a 1 in the FAB assay within the 4 treatment groups (n = 3 per group). Data are shown as means ± SE. IT, Immunotherapy; OM, omalizumab. Journal of Allergy and Clinical Immunology 2007 120, 688-695DOI: (10.1016/j.jaci.2007.05.034) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 5 Changes in allergen-specific IgG4 levels relative to pretreatment levels over the course of the study. Allergen-specific IgG4 levels were measured by means of ELISA. Individual data are shown, and differences between baseline measurements and time points were assessed by using the Friedman nonparametric repeated-measures test (∗P < .05, ∗∗P < .01, and ∗∗∗P < .001). IT, Immunotherapy; OM, omalizumab. Journal of Allergy and Clinical Immunology 2007 120, 688-695DOI: (10.1016/j.jaci.2007.05.034) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 6 Assessment of serum inhibitory properties 18, 30, and 42 weeks after discontinuation of treatment. Inhibitory activity of the serum was tested in the FAB assay, where binding of indicator serum alone was normalized to 100%. Data are shown as means ± SE. Differences between groups were assessed by using the Mann-Whitney U test. P, Placebo; IT, immunotherapy; OM, omalizumab. Journal of Allergy and Clinical Immunology 2007 120, 688-695DOI: (10.1016/j.jaci.2007.05.034) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions