Experimental protocols.

<p>Grey shaded boxes indicate periods of oral peanut exposure. Mothers were exposed to ground peanut for 4 weeks during either in the pre-conception period only (PC), during preconception, pregnancy and lactation (PC+PG+LC); or were not fed peanut, serving as controls (None). All the offspring underwent oral sensitization to peanut by being gavage-fed crude peanut extract (CPE) with cholera toxin or were tolerized by being gavage-fed CPE without adjuvant. In some experiments, young mice born to naïve mothers were exposed to the milk of naïve (naïve milk) or peanut-exposed (immune milk) mice concurrently with CPE and cholera toxin to induce peanut sensitization or CPE alone to induce oral tolerance.</p

Offspring’s response to peanut sensitization based on maternal feeding of peanut.

<p>(A) Experimental protocol. Serum antibodies (B-E) and an anaphylactic response to the peanut challenge as indicated by the elevated score during the oral (PO) challenge (F) (baseline symptom score 0 in all the mice), the lowered body temperature during the intraperitoneal (IP) challenge (G), and after the peanut sensitization were assessed in offspring born to mothers exposed to peanut preconceptionally, who either continued to feed peanut during pregnancy and lactation (PC+PG+LC) and those who did not (PC). Offspring born to mothers never exposed to peanut served as controls (None). Naïve, non-sensitized mice served as controls. Shown is a mean with SEM. >15 mice were used per group.</p

OVA epicutaneous sensitization after feeding OVA with or without murine milk.

<p>(A) Experimental protocol. Offspring come from naïve mothers. Offspring’s response to OVA challenge (B-C) and antibody responses to OVA sensitization (D-F). Mice post-weaning at 3–4 weeks of age were fed OVA +/- OVA-immune or naïve milk for 5 days followed by cutaneous sensitization with OVA. Mice immunized without prior OVA feeding (Unfed) were used as controls. X axis for D-F shows the dilution factor for the serum. Each group has 5 mice. Shown is a mean with SEM. **, p<0.01, ***, p<0.001, ****, p<0.00001 when compared to unfed.</p

Maternal feeding of peanut does not interfere with the induction of oral tolerance in offspring.

<p>(A) Experimental protocol. (B-D) Serum IgE antibodies after peanut immunization were assessed in offspring who were either fed crude peanut extract (CPE) orally prior to immunization with CPE (Fed) or not fed (Unfed). Offspring were either born to mothers exposed to peanut preconceptionally who either continued to feed peanut during pregnancy and lactation (PC+PG+LC) or did not (PC). Offspring born to mothers that were never exposed to peanut served as controls (None). In (C), although antibody levels are similar, the dilution curves are significantly different. Shown is a mean with SEM. Each group has 10–18 mice *, p<0.05, ****, p<0.0001.</p

Intestinal peanut uptake to the Peyer’s patches.

<p>Mice were gavage-fed FITC-labeled CPE in the presence of naïve murine milk or immune milk. Peyer’s patches were collected 10 min later. A-B show representative sections of immunofluorescence. FAE, follicle associated epithelium; SED, subepithelial dome. FITC, CPE; Magenta, CD11c+. C) Number of DCs staining positive for CPE per patch. **, p<0.01 when compared to immune milk.</p

Mothers’ preconception serum (A-B) and breast milk (C-D) antibody levels in mothers who were fed peanut preconceptionally (PC) and in those who were not (None).

<p>Presensitization antibody levels at 5 weeks of age are shown (E-F) for offspring born to peanut-exposed mothers who either continued to feed peanut throughout pregnancy and lactation (PG+LC) and in those who did not (PC). The dilution curves are not shown due to the small amount of serum available. Specific IgE was undetectable (data not shown). Titles indicate sample dilution. Shown is a mean (with SEM). 5–10 mothers per group and all their offspring (>15/group) were used for each experiment. *, p<0.05; **, p<0.01; ***, p<0.001 compared to None.</p