Grade of concordance of specific IgE (sIgE) in serum and Basophil activation test (BAT).

<p>Statistically significant differences were defined as *P<0.05, **P<0.01 and ***P<0.001.</p><p>Grade of concordance of specific IgE (sIgE) in serum and Basophil activation test (BAT).</p

Patient characteristics.

<p>Patient characteristics.</p

Analysis of patients negative to rVes v 5 with rVes v 3, rVes v 1 and natural venom, improves the trade-off between sensitivity and specificity of the Basophil activation test.

<p>(<b>A</b>) Percentage of patients positive to basophil activation test performed with natural venom and recombinant allergens. (<b>B</b>) Analysis of patients negative to recombinant Ves v 5 with natural venom and other recombinant allergens. (<b>C</b>) Specificity and sensitivity of natural venom, recombinant Ves v 5 and recombinant Ves v 3 at cut off 15.7% CD63 expression. (<b>D</b>) Specificity and sensitivity of multistep analysis of BAT considering rVes v 5 alone, rVes v 5 plus rVes v 3, and rVes v 5 plus rVes v 3 plus natural venom.</p

Recombinant allergens differentially recognize venom allergic patients.

<p><i>Table legend</i>: nVenom =  natural venom, V1 =  rVes v 1 V2 =  rVes v 2 V3 =  rVes v 3, V5 =  rVes v 5; “+” =  positive patient; “–“ =  negative patient; N.D. =  not done: sIgE =  specific IgE; BAT =  Basophil activation test. Cut-offs: 0.35 KU/L ImmunoCaps, 0.1 O.D. ELISAs, 15.7% CD63 expression in BAT. Empty spaces indicate untested samples.</p><p>Recombinant allergens differentially recognize venom allergic patients.</p

Wasp venom extracts activate basophils in the whole blood of allergic patients.

<p>(<b>A</b>) Identification of basophils according to dimension and IgE levels. (<b>B</b>) Representative two-color flow cytometric staining dot plots of IgE and CD63 on basophils unstimulated, stimulated with anti-FCεRI and nVenom. (<b>C</b>) CD63 expression on basophils in absence of stimulation, stimulated with anti-FCεRI and nVenom. Scale bars indicate mean ± SD.</p

Recombinant allergens strongly activate basophils in allergic subjects.

<p>Representative two-color flow cytometric staining dot plots of IgE and CD63 on basophils of (<b>A</b>) five representative allergic subjects responsive to each recombinant allergen, (<b>B</b>) a representative multi-sensitized patient responsive to all the recombinant allergens (<b>C</b>) degree of basophil activation in a representative subject of the control group.</p

Recombinant allergens improve the <i>in vitro</i> test specificity for the identification of wasp venom allergic patients.

<p>ROC curves of specific IgE measurement (sIgE) of recombinant Ves v 1, Ves v 2 and Ves v 3 by ELISA (O.D.) (<b>A–C</b>) and recombinant Ves v 5 by ImmunoCAP (KU/L) (<b>D</b>). ROC curves of CD63% trade-off of basophil stimulated with recombinant Ves v 1, Ves v 2, Ves v 3 and Ves v 5 (<b>E–H</b>).</p