Cross-protection of AIT-induced antibodies to related allergens requires a high degree of structural identity

Background: In contrast to sublingual immunotherapy (SLIT) with recombinant Mal d 1 (rMal d 1-SLIT), SLIT with rBet v 1 (rBet v 1-SLIT) induced Mal d 1-cross-reactive antibodies without IgE-blocking activity. To elucidate whether the development of cross-protective IgG responses depends on the degree of molecular identity of allergens we compared the cross-reactivity, cross-blocking activity, and affinity of SLIT-induced antibodies with allergens of varying amino acid sequence identities to Bet v 1 and Mal d 1, namely Cor a 1.04 (hazelnut), Pru av 1 (cherry), and Dau c 1 (carrot).

Methods: Allergen-specific antibodies were quantified by ELISA. IgE blocking was analyzed by inhibition of allergen-induced basophil activation and IgE-facilitated allergen-presentation to T cells. The affinity of SLIT-induced antibodies was studied by acidic dissociation ELISA and competition ELISA. Identical surface areas on allergens were predicted using an in-house designed script based on structural alignments.

Results: rBet v 1-SLIT-induced IgG antibodies cross-reacted with all allergens except Dau c 1. rMal d 1-SLIT-induced antibodies predominantly cross-reacted with Pru av 1 and displayed significantly higher IgE blocking to Pru av 1 than rBet v 1-SLIT-induced antibodies. rMal d 1-SLIT-induced IgG1 showed higher affinity to Mal d 1 and Pru av 1. Surface analysis revealed 84% identical area on Mal d 1 and Pru av 1. Furthermore, we identified two surface areas potentially containing epitopes present on these allergens and absent on Bet v 1.

Conclusion: In summary, our findings suggest that a relatively high threshold of similarity is required to establish effective cross-blocking antibodies to related allergens. Apparently, the structural identity between Bet v 1 and Mal d 1 is below this threshold. Therefore, this study may explain why immunotherapy with birch pollen allergen often fails to reduce birch pollen-related apple allergy.