GM-maize consumption does not affect lung inflammation at the initiation or relapse of allergic asthma.

<p>Representative photomicrographs of lungs from A) Naïve mice fed a basal diet, and mice at disease initiation fed B) a basal diet, C) nGM-maize, or D) GM-maize, and mice at disease relapse fed, E) a basal diet, F) nGM-maize, or G) GM-maize. H&E-stained lung sections were taken with a 10x objective and are representative data from 2 independent experiments. Arrowheads indicate areas of inflammation and insets are at 40x and illustrate eosinophils within inflammatory infiltrates.</p

GM-maize consumption does not affect mucus secretion at the initiation or relapse of allergic asthma.

<p>Representative photomicrographs of lungs from A) Naïve mice fed a basal diet, and mice at disease initiation fed B) a basal diet, C) nGM-maize, or D) GM-maize, and mice at disease relapse fed, E) a basal diet, F) nGM-maize, or G) GM-maize. Lung PAS-stained sections were observed with a 10x objective. These are representative data from 2 independent experiments. Arrowheads indicate mucus within lung epithelial goblet cells.</p

Experimental protocols.

<p><i>Adjuvant effect of GM-maize on the initiation of OVA-induced allergic lung disease.</i> Groups of mice were fed either GM-maize, nGM-maize, or basal diets between days 0 and 32 of the initiation of allergic asthma. Mice were immunized to induce allergic disease with 10 µg of OVA on days 0 and 21. One week later, the mice were nebulized with OVA on days 28 and 29. On day 32, antibody titres and allergic lung responses were measured. <i>Adjuvant effect of GM-maize on OVA-induced allergic disease relapse.</i> Mice were induced with allergic disease on days 0 and 21 and aerosolized on days 28 and 29 and then allowed to recover for another month (all signs of acute lung and airway inflammation and mucus hypersecretion resolves). Between day 61 and 95, mice were then fed either GM-maize, nGM-maize or basal diets. On days 91 and 92, mice were rechallenged with aerosolized OVA to induce a disease relapse. On day 95, they were evaluated for antibody titres and allergic lung responses.</p

GM-maize consumption does not affect OVA-specific antibody production at the initiation or relapse of allergic asthma.

<p>OVA-specific antibody response induced by feeding in mice. Three days after the last aerosol challenge the level of OVA-specific IgG1, IgE and IgG2a antibody response induced by feeding of different diets. Individual serum samples were analyzed for IgG1 and IgE (Th2 phenotype) and IgG2a (Th1 phenotype). Data are expressed as mean O.D. ± SD, n = 8.</p

GM-maize consumption does not affect the severity of lung inflammation and the percentage of eosinophils in the lungs at the initiation or relapse of allergic asthma.

<p>A) Severity scores graded in H&E-stained lung sections and B) the percentage of eosinophils within infiltrates measured in Luna-stained lung sections from mice at disease initiation and relapse fed diets as shown. These are representative data from 2 independent experiments. Severity scores are reported as means ± SEM; n = 8. Groups are compared using the Kruskal-Wallis test ^*p<0.05 sample vs. naïve groups.</p

Ingredient composition and chemical content of experimental diets.

<p>*The basal diet ingredient composition is unavailable for commercial reasons. However, in descending order the ingredients are: ground wheat, soybean meal & soy protein concentrate, ground corn & middlings, oat middlings, corn gluten meal/feed, dried sugar beet pulp, mineral compounds, soybean oil, amino acids [DL-Met, L-Lys HCl], vitamins & trace elements.</p

GM-maize consumption does not influence the development of airway allergic inflammation at initiation or relapse of allergic asthma.

<p>Three days after the last aerosol challenge, BAL of individual mice was collected and differential cells counts in OVA-induced allergic asthma initiation and during disease relapse were performed. We compared naïve mice vs. mice fed mice basal, GM-maize, and nGM-maize diets for 32–34 days. Representative data from 2 independent experiments are presented as absolute counts of eosinophils, macrophages, neutrophils and lymphocytes and reported as means ± SEM; n = 8. The height of the columns is the total cell count. Kruskal-Wallis test ^*p<0.05 sample vs. naïve group followed by Dunn’s multiple comparison test.</p

GM-maize consumption does not affect the amount of mucus secreted at initiation or relapse of allergic asthma.

<p>Mucus scores graded on PAS-stained lung sections from mice at disease initiation and relapse fed diets as shown. These are representative data from 2 independent experiments. Mucus scores are reported as means ± SEM; n = 8. Groups were compared using the Kruskal-Wallis test ^*p<0.05 sample vs. naïve groups.</p

Induction of EAAD and treatment schedule.

<p>Schematic depiction of the experimental protocol used for the induction of EAAD, the application of treatments and the optical imaging performed.</p

Quantification of <i>in vivo</i> imaging results.

<p>Box plot of average fluorescence intensities over the lung area for all groups at 48-labeled anti-Siglec-F antibody injection. Lung intensities of EAAD mice are significantly higher (represented by asterisk *) compared with control mice and treated mice at 48 h and 72 h after antibody application; A = EAAD, C = control, AD = EAAD, dexamethasone treated, AE = EAAD, beta-escin treated.</p