Rationally Engineered Self-Assembling Protein Nanoparticles for Allergen-Specific Peanut Allergy Immunotherapy

Abstract

Peanut allergy is a prevalent immunoglobulin E (IgE)-mediated food allergy that can cause life-threatening anaphylaxis. Current treatment options provide limited efficacy and variable durability of protection. While allergen-displaying virus-like particles (VLPs) have shown promise, their development is often constrained by limited design flexibility and manufacturing complexity. Here, we develop self-assembling protein nanoparticles (SApNPs) displaying the major allergens Ara h 2 and Ara h 6 as a modular platform for peanut allergy immunotherapy. Using the 60-mer TIP60 scaffold, we engineer nanoparticles by expanding particle size through -helical insertion and replacing the human myosin-X segment with evolutionarily distant catfish homologs, followed by structure-guided and evolution-inspired optimization to enhance thermostability while minimizing human sequence content. Formulated with a Toll-like receptor 9 (TLR9) agonist, these allergen-displaying SApNPs confer robust protection against peanut-induced anaphylaxis in both prophylactic and therapeutic settings. Mechanistically, SApNPs exhibit prolonged lymph node retention, sustained presentation on follicular dendritic cells, and enhanced germinal center responses compared with soluble allergens, promoting allergen-specific IgG and functional T cell responses while limiting IgE escalation. Collectively, these findings establish allergen-displaying SApNPs as a rationally engineered, scalable, and modular platform for peanut allergy treatment with potential applicability to other allergic diseases.