Additional file 1: of Acute and preventive management of anaphylaxis in German primary school and kindergarten children

Excerpt of the questionnaire for teachers/child-care providers about anaphylactic reactions in children (questions not shown concerned demographic background). (DOCX 21 kb

Additional file 2: of Acute and preventive management of anaphylaxis in German primary school and kindergarten children

List of items that were included in questionnaire for parents about anaphylactic reactions in children. (DOCX 26 kb

Demonstration of autoantibody specificity by inhibition in ELISA.

<p>A: Inhibition of serum IgA reactivity to glycoprotein 2 (GP2). A serum with high anti-GP2 IgA level at a dilution giving an optical density (OD) of 0.8 was pre-incubated with recombinant GP2 (Isoform alpha) and tissue transglutaminase (TG) at decreasing concentrations from 0–10 μg/mL. B: Inhibition of serum IgA reactivity to tTG. A serum with high anti-tTG IgA level at a dilution giving an optical density (OD) of 1.5 was pre-incubated with recombinant GP2 (Isoform alpha) and tTG at decreasing concentrations from 0–10 μg/mL. Data are presented as means of triplicate measurements with the corresponding standard deviation.</p

Association between anti-GP2 IgA level and degree of villous atrophy.

<p>IgA antibody levels to glycoprotein 2 (GP2) determined in 153 patients with active celiac disease are associated with the grade of villous atrophy according to Marsh classification. The dotted line indicates cut-off value of 20 U/mL for anti-GP2 IgA. Data are displayed as U/mL in Box-and-Whisker plots with far out values, defined as values that are smaller than the lower quartile minus 3 times the interquartile range, or larger than the upper quartile plus 3 times the interquartile range, displayed as solid triangles. * p = 0.001, ** p = 0.002.</p

Loss and Gain of Tolerance to Pancreatic Glycoprotein 2 in Celiac Disease

<div><p>Background</p><p>Autoantibodies against pancreatic secretory-granule membrane glycoprotein 2 (GP2) have been demonstrated in patients with Crohn’s disease but recently also with celiac disease (CD). Both entities are characterized by intestinal barrier impairment with increased gut permeability. Pathophysiological hallmark of CD is a permanent loss of tolerance to alimentary gliadin and a transient loss of tolerance to the autoantigen human tissue transglutaminase (tTG). Therefore, we explored the behavior of loss of tolerance to GP2 reported in CD.</p><p>Methods</p><p>We assessed prevalences and levels of autoantibodies against GP2, CD-specific antibodies to endomysial antigens and tTG as well as Crohn’s disease-specific anti-<i>Saccharomyces cerevisiae</i> antibodies in sera of 174 patients with active CD, 84 patients under gluten-free diet (GFD) and 129 controls. Furthermore, we looked for an association between anti-GP2 antibody positivity and degree of mucosal damage in CD.</p><p>Results</p><p>We found significantly elevated anti-GP2 IgA positivity in active CD patients (19.5%) compared to CD patients under GFD (0.0%) and controls (5.4%, p < 0.001, respectively). Anti-GP2 IgA levels correlated significantly with CD-specific antibodies (p < 0.001). Anti-GP2 autoantibody positivity disappeared under GFD similarly to CD-specific autoantibodies against tTG and endomysial antigens. For the first time, IgA antibody levels to GP2 are demonstrated to be associated with degree of villous atrophy according to Marsh classification.</p><p>Conclusions</p><p>Anti-GP2 IgA seems to be associated with disease activity in a distinct subgroup of patients with CD. The observed loss of tolerance to GP2 in a subset of patients with CD is transient and disappears under GFD.</p></div

Antibody kinetics in 4 celiac disease patients.

<p>Antibody kinetics in 4 celiac disease patients (A-D) with at least two consecutive samples before and after the onset of a gluten free diet. In fact, in all patients anti- glycoprotein 2 (GP2) IgA levels were reduced to values below the cut-off. Interestingly, in one patient (patient B) with co-existing type 1 diabetes who became positive for anti-GP2 IgA in parallel with anti- tissue transglutaminase (tTG) IgA, anti-GP2 IgA turned also negative under gluten free diet.</p

Correlation of anti-GP2 IgA with celiac disease (CD) specific antibodies.

<p>Association of anti-GP2 IgA with endomysial antibodies (EmA) (A) and anti- tissue transglutaminase (tTG) IgA (B) in 174 patients with active CD. (Spearman's coefficient of rank correlation for (A) 0.466 and (B) 0.494 (p<0.001, respectively).</p

Clinical characteristics of the study groups.

<p>^a In this group, 51 patients of group 1 with active CD and follow-up serology under gluten-free diet (GFD) are included.</p><p>^b For the other 21 patients, the original histology was not available. In one patient diagnosis was done according to the new ESPGHAN guidelines without biopsy [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0128104#pone.0128104.ref018" target="_blank">18</a>].</p><p>^c The 36 patients with family history of CD were from 23 different families.</p><p>celiac disease (CD), gluten-free diet (GFD).</p><p>Clinical characteristics of the study groups.</p

Descriptive statistics of the repeated quantification (number of repetitions: 20).

<p>Descriptive statistics of the repeated quantification (number of repetitions: 20).</p

MRI images of the skull with different contrasts.

<p>Exemplarily, a specimen of group 2 after six weeks healing time was chosen. The yellow rectangle highlights the region of interest which includes the artificial defect. The arrows indicate the bone substitute material. The scale bars represent 1.0 mm. The repetition time and echo time which have been used for MR imaging (cf. <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0179249#pone.0179249.t002" target="_blank">Table 2</a>) were abbreviated with T_R and T_E, respectively. The label description can be also found in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0179249#pone.0179249.t003" target="_blank">Table 3</a>. (A) Proton density images which showed the best signal-to-noise ratio. They were used for quantitative MRI. (B) T₂ weighted images highlight tissues with high content of unbound water. Adipose tissue appears bright, too. (C) With T₁ weighted images the adipose tissue can be identified.</p