Iván López-Expósito, PhD, Ying Song, MD, Kirsi M

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Maternal peanut exposure during pregnancy and lactation reduces peanut allergy risk in offspring Iván López-Expósito, PhD, Ying Song, MD, Kirsi M. Järvinen, MD, PhD, Kamal Srivastava, MPhil, Xiu-Min Li, MD Journal of Allergy and Clinical Immunology Volume 124, Issue 5, Pages 1039-1046 (November 2009) DOI: 10.1016/j.jaci.2009.08.024 Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 Experimental protocols. A, Intragastrically sensitized PN-allergic female mice were mated after a 5 weeks PN sensitization and one week boosting. B, In another set of experiments, during pregnancy and lactation mice were divided into 2 groups: (PNA-M/None; n = 5) and (PNA-M/PN; n = 5). A naive group (n = 5) was employed as a control. Five-week-old offspring from each group were then challenged with 200 mg PN/mouse. C, In a third set of experiments, five-week-old offspring from each group were sensitized weekly during 6 weeks (10 mg PN+CT/mouse). Journal of Allergy and Clinical Immunology 2009 124, 1039-1046DOI: (10.1016/j.jaci.2009.08.024) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 A, Symptoms of anaphylaxis. B, Temperatures. Bars indicate the median of 15 offspring from PNA-M and 12 offspring in the naive group. ∗∗∗P < .001 vs naive. C, Offspring from PNA-M experiencing redness and swelling around the mouth in comparison with a naive mouse. D, Signs of intestinal anaphylaxis; note edema in the small intestine of an offspring of PNA-M in comparison with a naive mouse intestine. Journal of Allergy and Clinical Immunology 2009 124, 1039-1046DOI: (10.1016/j.jaci.2009.08.024) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 3 Mechanisms of peanut-induced systemic anaphylaxis in offspring of PNA-Ms. A, Thirty minutes before challenge, mice were pretreated with either intraperitoneal injection of cyproheptadine or intravenous injection of CV-3988. Symptoms of anaphylaxis were scored as described in the Methods. B, Temperatures were measured 30 minutes after challenge with a rectal probe digital thermometer. Bars indicate the median of 5 mice per group. ∗P < .05, ∗∗∗P < .001 vs PNA-M. Journal of Allergy and Clinical Immunology 2009 124, 1039-1046DOI: (10.1016/j.jaci.2009.08.024) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 4 Humoral and cellular response in offspring of PNA-Ms and naive mothers. A, Serum peanut (PN)-specific IgG1 (μg/mL). B, Serum PN-specific IgG2a (μg/mL). Data are expressed as means ± SEMs for each group (n = 13 for PNA-M group and n = 5 for naive). ∗∗∗P < .001 vs naive. PCA reactions after injection of unheated (C), heated (D), and naive (E) sera. Journal of Allergy and Clinical Immunology 2009 124, 1039-1046DOI: (10.1016/j.jaci.2009.08.024) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 5 A, Serum peanut (PN)-specific IgG2a (ng/mL). B, Serum PN-specific IgG1 (ng/mL). C, Ratio between PN-specific IgG2a/IgG1. (D) IL-4, (E) IL-5, (F) IL-13, (G) IL-17, (H) IFN-γ, (I) TGF-β, and (J) IL-10 levels in culture supernatants (pg/mL). Data in A-J are expressed as means ± SEMs of duplicates of ELISA for each group (n = 7). ∗P < .05, ∗∗P < .01, ∗∗∗P < .001 vs PNA-M/none; #P < .05, ##P < .01, ###P < .001 vs naive. Journal of Allergy and Clinical Immunology 2009 124, 1039-1046DOI: (10.1016/j.jaci.2009.08.024) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 6 Humoral response of offspring of PNA-M/none and PNA-M/PN. A, Serum peanut (PN)-specific IgE (ng/mL). B, Serum PN-specific IgG1 (ng/mL) and (C) Serum PN-specific IgG2a (ng/mL) were evaluated by antigen-specific ELISA. Data are expressed as means ± SEMs of triplicates for each group (pooled samples; n = 5 for PNA-M/none group and n = 10 for PNA-M/PN). ∗∗P < .01, ∗∗∗P < .001 vs PNA-M/PN. Journal of Allergy and Clinical Immunology 2009 124, 1039-1046DOI: (10.1016/j.jaci.2009.08.024) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions