Flow cytometric analysis of in vitro activated basophils, specific IgE and skin tests in the diagnosis of pollen-associated food allergy.

Background: Specific immunoglobulin E (IgE) and commercially available skin prick tests have been demonstrated to be unreliable methods to diagnose pollen-associated food allergy. To evaluate the predictive value of the basophil activation test (BAT) in pollen-associated food allergy, the apple-mediated oral allergy syndrome (OAS) in patients with birch pollinosis was chosen as a representative model.
Methods: Patients with birch pollen allergy and a history of apple-mediated OAS (OAS(+), n = 29), patients with birch allergic without OAS (OAS(-), n = 22), and healthy controls (HC, n = 10) without birch pollen allergy and OAS were included. Apple IgE was quantified by the CAP FEIA method. Skin prick tests were performed with a Jonagold apple extract. Flow cytometric analysis of basophils activated with the same Jonagold extract was based on double staining with anti-IgE/anti-CD63 monoclonal antibodies.
Results: Comparison between OAS(+) subjects and HC showed sensitivities and specificities of 96% and 100% for apple IgE and 88% and 100% for the apple skin prick test, respectively. For the BAT, sensitivity and specificity were 100%. In contrast, when nonresponders on the BAT were considered, sensitivity decreased to 90%. In a separate analysis between OAS(+) and OAS(-) subjects, specificities decreased to 30% for apple IgE and to 80% for the apple skin test, respectively. The BAT reached a sensitivity of 88% and a specificity of 75%.
Conclusion: Flow cytometry-assisted quantification of in vitro basophil activation seems to be a reliable instrument in the diagnosis of this model of pollen-associated food allergy. In addition, this study reemphasizes that the specificity of diagnostic allergy tests decreases considerably when, apart from HC, control individuals with cross-reactive antibodies are included.