Chimeras of Bet v 1 and Api g 1 reveal heterogeneous IgE responses in patients with birch pollen allergy.

Background Characterization of IgE-binding epitopes of allergens and determination of their patient-specific relevance is crucial for the diagnosis and treatment of allergy. Objective We sought to assess the contribution of specific surface areas of the major birch pollen allergen Bet v 1.0101 to binding IgEof individual patients. Methods Four distinct areas of Bet v 1 representing in total 81% of its surface were grafted onto the scaffold of its homolog, Api g 1.0101, to yield the chimeras Api-Bet-1 to Api-Bet-4. The chimeras were expressed in Escherichia coli and purified. IgEbinding of 64 sera from Bet v 1–sensitized subjects with birch pollen allergy was determined by using direct ELISA. Specificity was assessed by means of inhibition ELISA. Results rApi g 1.0101, Api-Bet-1, Api-Bet-2, Api-Bet-3, and Api-Bet-4 bound IgEfrom 44%, 89%, 80%, 78%, and 48% of the patients, respectively. By comparing the amount of IgEbinding to the chimeras and to rApi g 1.0101, 81%, 70%, 75%, and 45% of the patients showed significantly enhanced IgEbinding to Api-Bet-1, Api-Bet-2, Api-Bet-3, and Api-Bet-4, respectively. The minority (8%) of the sera revealed enhanced IgEbinding exclusively to a single chimera, whereas 31% showed increased IgEbinding to all 4 chimeras compared with rApi g 1.0101. The chimeras inhibited up to 70% of IgEbinding to rBet v 1.0101, confirming the specific IgErecognition of the grafted regions. Conclusion The Bet v 1–specific IgEresponse is polyclonal, and epitopes are spread across the entire Bet v 1 surface. Furthermore, the IgErecognition profile of Bet v 1 is highly patient specific.