Frequencies of allergen-specific IgE reactivities in the AD and SE patients as tested by MeDALL allergen chip, Immunoblotting, ImmunoCAP^™ and RAST-based dot-blot assay.

<p>Frequencies of allergen-specific IgE reactivities in the AD and SE patients as tested by MeDALL allergen chip, Immunoblotting, ImmunoCAP^™ and RAST-based dot-blot assay.</p

Summary of allergen-specific IgE reactivities in the AD patients.

<p>Summary of allergen-specific IgE reactivities in the AD patients.</p

Demographic and clinical characterization of AD patients and controls.

<p>Demographic and clinical characterization of AD patients and controls.</p

The culprit insect but not severity of allergic reactions to bee and wasp venom can be determined by molecular diagnosis

<div><p>Background</p><p>Allergy to bee and wasp venom can lead to life-threatening systemic reactions. The identification of the culprit species is important for allergen-specific immunotherapy.</p><p>Objectives</p><p>To determine a panel of recombinant bee and wasp allergens which is suitable for the identification of bee or wasp as culprit allergen sources and to search for molecular surrogates of clinical severity of sting reactions.</p><p>Methods</p><p>Sera from eighty-seven patients with a detailed documentation of their severity of sting reaction (Mueller grade) and who had been subjected to titrated skin testing with bee and wasp venom were analyzed for bee and wasp-specific IgE levels by ImmunoCAP^TM. IgE-reactivity testing was performed using a comprehensive panel of recombinant bee and wasp venom allergens (rApi m 1, 2, 3, 4, 5 and 10; rVes v 1 and 5) by ISAC chip technology, ImmunoCAP and ELISA. IgG₄ antibodies to rApi m 1 and rVes v 5 were determined by ELISA and IgE/IgG₄ ratios were calculated. Results from skin testing, IgE serology and IgE/IgG₄ ratios were compared with severity of sting reactions.</p><p>Results</p><p>The panel of rApi m 1, rApi m 10, rVes v 1 and rVes v 5 allowed identification of the culprit venom in all but two of the 87 patients with good agreement to skin testing. Severities of sting reactions were not associated with results obtained by skin testing, venom-specific IgE levels or molecular diagnosis. Severe sting reactions were observed in patients showing < 1 ISU and < 2kU_A/L of IgE to Api m 1 and/or Ves v 5.</p><p>Conclusion</p><p>We identified a minimal panel of recombinant bee and wasp allergens for molecular diagnosis which may permit identification of bee and/or wasp as culprit insect in venom-sensitized subjects. The severity of sting reactions was not associated with parameters obtained by molecular diagnosis.</p></div

Pie charts showing the contribution of (A), allergen sources and (B), individual allergen components to IgE sensitization.

<p>Each chart represents 100% of IgE reactivities detected in plasma from all AD patients of the respective group, severe AD (left chart) and moderate AD (right chart), in (<b>A)</b> to six allergen sources using the MeDALL allergen-chip (ISU ≥ 0.3) and to extracts of <i>M</i>. <i>sympodialis</i>, <i>S</i>. <i>aureus</i> and the human cell line A431 using immunoblotting, and in (<b>B)</b> to 25 allergen molecules using the MeDALL allergen-chip (ISU ≥ 0.3). The sizes of the segments represent the proportion of the respective in (<b>A)</b>, allergen source and in (<b>B)</b>, allergen molecule among all recognized. Allergen sources/molecules start a 12 o’clock of the pie chart and continue clock-wise as listed with the color code.</p

Development of allergen-specific IgE levels.

<p>Relative changes of IgE-levels (y-axes) to Phl p 5, Bet v 1 and Phl p 1 (top to bottom: right labels of each chart) compared to t1 (day 1) are shown for all visits (t2: day 8; t3: day 15; t4: day 29; t5: day 43; t6: day 57, x-axes). Results for the steroid-treated group are shown in blue, the placebo-group in green. Results from participants challenged with Phl p 5 are depicted in the left column, those for the Bet v 1-challenged group on the right side. Outliers that lie between 1.5 and 3 times the interquartile range below the first or above the third quartile are shown as open circles (“o”), those that lie beyond 3 times the interquartile range are depicted by asterisks (“*”).There were no significant differences between the fluticasone and placebo groups at any time point.</p

Development of Phl p 5-specific antibody responses.

<p>Percentage changes of Phl p 5-specific IgG₁, IgG₂, IgG₄, IgA and IgM antibody levels (y-axes) at the day of the first nasal provocation (0) and thereafter (x-axes). Uninterrupted lines: Patients with steroid spray; dotted lines: Patients with placebo spray; black squares: provocation with rPhl p 5; grey dots: provocation with rBet v 1.</p

Distribution of study subjects.

<p>Of the 48 volunteers who were randomized, 45 completed the trial. Drop-outs are marked in yellow. INCS: intranasal corticosteroid; NPT: nasal provocation test; URTI: upper respiratory tract infection; i.m.: intra-muscular</p

Effect of fluticasone versus placebo on the median relative change in allergen-specific IgE levels.

<p>The ratio of relative changes from t1 is the relative change in the fluticasone group divided by the relative change in the placebo group. A ratio of 1 corresponds to no differential effect between fluticasone and placebo, a ratio below 1 corresponds to a beneficial blunting effect of fluticasone on allergen-induced IgE levels. 95% CI low and 95% CI up indicate the lower or upper bound of a 95% confidence interval, respectively.</p><p>Effect of fluticasone versus placebo on the median relative change in allergen-specific IgE levels.</p

Association of venom extract or allergen-specific IgE levels and severity of sting reactions.

<p>Specific IgE levels (medians: horizontal lines) to (A) bee and (B) wasp venom (kU_A/L), (C) rApi m 1 (kU_A/L), (D) rApi m 1 (ISU), (E) rVes v 5 (kU_A/L), (F) rVes v 5 (ISU) were plotted against the severities of sting reactions (x-axes: Mueller grade) for Slovenian and German patients with identified culprit insect.</p