Design of a heterotetravalent synthetic allergen that reflects epitope heterogeneity and IgE antibody variability to study mast cell degranulation

作者：Handlogten Michael W; Kiziltepe Tanyel; Bilgicer Basar^*

来源：Biochemical Journal, 2013, 449(1): 91-99.

DOI：10.1042/BJ20121088

• allergy
• IgE antibody
• heterotetravalent
• mast cell degranulation
• multivalency
• synthetic allergen

The present paper describes the design of a HtTA (heterotetravalent allergen) as a multi-component experimental system that enables an integrative approach to study mast cell degranulation. The HtTA design allows presentation of two distinct haptens, each with a valency of 2, thereby better reflecting the complexity of natural allergens by displaying epitope heterogeneity and IgE antibody variability. Using the HtTA design, synthetic allergens HtTA-1 and HtTA-2 were synthesized to model a combination of epitope/IgE affinities. HtTA-1 presented DNP (2,4-dinitrophenyl) and dansyl haptens (K-d = 22 and 54 nM for IgE(DNP) and IgE(dansyl) respectively) and HtTA-2 presented dansyl and the weak-affinity DNP-Pro (DNP-proline) haptens (K-d = 550 nM for IgE(DNP)). Both HtTAs effectively induced degranulation when mast cells were primed with both IgE(DNP) and IgE(dansyl) antibodies. Interestingly tetravalent DNP-Pro or bivalent dansyl were insufficient in stimulating a degranulation response, illustrating the significance of valency, affinity and synergy in allergen-IgE interactions. Importantly, maximum degranulation with both HtTA-1 and HtTA-2 was observed when only 50% of the mast cell-bound IgEs were hapten-specific (25 % IgE(dansyl) and 25 % IgE(DNP)). Taken together, results of the present study establish the HtTA system as a physiologically relevant experimental model and demonstrates its utility in elucidating critical mechanisms of mast cell degranulation.

• 出版日期2013-1-1