Figure 1.
Wasp venom extracts activate basophils in the whole blood of allergic patients.
(A) Identification of basophils according to dimension and IgE levels. (B) Representative two-color flow cytometric staining dot plots of IgE and CD63 on basophils unstimulated, stimulated with anti-FCεRI and nVenom. (C) CD63 expression on basophils in absence of stimulation, stimulated with anti-FCεRI and nVenom. Scale bars indicate mean ± SD.
Figure 2.
Recombinant allergens strongly activate basophils in allergic subjects.
Representative two-color flow cytometric staining dot plots of IgE and CD63 on basophils of (A) five representative allergic subjects responsive to each recombinant allergen, (B) a representative multi-sensitized patient responsive to all the recombinant allergens (C) degree of basophil activation in a representative subject of the control group.
Table 1.
Patient characteristics.
Figure 3.
Recombinant allergens improve the in vitro test specificity for the identification of wasp venom allergic patients.
ROC curves of specific IgE measurement (sIgE) of recombinant Ves v 1, Ves v 2 and Ves v 3 by ELISA (O.D.) (A–C) and recombinant Ves v 5 by ImmunoCAP (KU/L) (D). ROC curves of CD63% trade-off of basophil stimulated with recombinant Ves v 1, Ves v 2, Ves v 3 and Ves v 5 (E–H).
Table 2.
Grade of concordance of specific IgE (sIgE) in serum and Basophil activation test (BAT).
Table 3.
Recombinant allergens differentially recognize venom allergic patients.
Figure 4.
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.
(A) Percentage of patients positive to basophil activation test performed with natural venom and recombinant allergens. (B) Analysis of patients negative to recombinant Ves v 5 with natural venom and other recombinant allergens. (C) Specificity and sensitivity of natural venom, recombinant Ves v 5 and recombinant Ves v 3 at cut off 15.7% CD63 expression. (D) 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.