Grass pollen immunotherapy: IL-10 induction and suppression of late responses precedes IgG4 inhibitory antibody activity James N. Francis, PhD, Louisa.

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Grass pollen immunotherapy: IL-10 induction and suppression of late responses precedes IgG4 inhibitory antibody activity James N. Francis, PhD, Louisa K. James, PhD, Giannis Paraskevopoulos, MD, Cheukyee Wong, BSc, Moises A. Calderon, MD, PhD, Stephen R. Durham, MD, Stephen J. Till, MD, PhD Journal of Allergy and Clinical Immunology Volume 121, Issue 5, Pages 1120-1125.e2 (May 2008) DOI: 10.1016/j.jaci.2008.01.072 Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 Grass pollen immunotherapy results in reduced symptoms and decreased conjunctival reactivity to allergen. Eighteen patients were randomized to receive active immunotherapy (IT; n = 12) or placebo injections (n = 6) for 1 year. A, Overall assessment of symptoms was recorded on a numeric scale (−3 to +3) by each patient at the end of the study. B, Conjunctival provocation testing was performed at the end of the study. Journal of Allergy and Clinical Immunology 2008 121, 1120-1125.e2DOI: (10.1016/j.jaci.2008.01.072) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 Time-course analysis of clinical measurements during the first year of grass pollen immunotherapy. The dose of grass pollen allergen administered at each immunotherapy visit is represented in A. Early (B) and late (C) skin responses were assessed at 15 minutes and 24 hours after intradermal challenge with grass pollen allergen. Data are normalized to time after initial injection for the first 8 weeks of treatment, and thereafter data are plotted against collection date to assess potential effects of the grass pollen season. Data are expressed as means ± SEs and analyzed by using ANOVA with the Bonferroni correction. ∗Primary significance from baseline. EPR, Early-phase response; LPR, late-phase response. Journal of Allergy and Clinical Immunology 2008 121, 1120-1125.e2DOI: (10.1016/j.jaci.2008.01.072) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 3 Time-course analysis of serum antibody measurements during the first year of grass pollen immunotherapy. Allergen-specific IgG4 (A) and IgA (B) levels were measured in serum by means of ELISA, and IgE was measured by using RAST (C). Data are normalized to time and analyzed as previously. ∗Primary significance from baseline. Journal of Allergy and Clinical Immunology 2008 121, 1120-1125.e2DOI: (10.1016/j.jaci.2008.01.072) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 4 Time-course analysis of serum inhibitory activity and IL-10 production during the first year of grass pollen immunotherapy. Serum was tested for inhibition of FcεRI-mediated allergen-induced basophil histamine release in whole blood, with histamine being measured by means of ELISA (A). Serologic inhibitory activity against FcεRII/CD23-dependent IgE-facilitated allergen binding (B) was measured by using the IgE-facilitated allergen binding assay (FAB). IL-10 production by PBMCs stimulated with grass pollen allergen was measured by means of ELISA (C). Data are normalized to time and analyzed as previously described. ∗Primary significance from baseline. Journal of Allergy and Clinical Immunology 2008 121, 1120-1125.e2DOI: (10.1016/j.jaci.2008.01.072) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions