Allergen-specific immunotherapy (AIT) is a desensitizing treatment for allergic diseases that corrects the underlined pathological immune response to innocuous protein antigens, called allergens. Recombinant allergens employed in the AIT allowed the production of well-defined formulations that possessed consistent quality but were often less efficient than natural allergen extracts. Combining recombinant allergens with an adjuvant or immunomodulatory agent could improve AIT efficacy. This study aimed to perform structural and functional characterization of newly designed recombinant chimera composed of the Bet v 1, the major birch pollen allergen, and Banana Lectin (BanLec), TLR2, and CD14 binding protein, for the application in AIT. rBet v 1-BanLec chimera was designed in silico and expressed as a soluble fraction in Escherichia coli. Purified rBet v 1-BanLec (33.4 kDa) retained BanLec-associated biological activity of carbohydrate-binding and preserved IgE reactive epitopes of Bet v 1. The chimera revealed secondary structures with predominant β sheets. The immunomodulatory capacity of rBet v 1-BanLec tested on macrophages showed changes in myeloperoxidase activity, reduced NO production, and significant alterations in the production of cytokines when compared to both rBanLec and rBet v 1. Comparing to rBet v 1, rBet v 1-BanLec was demonstrated to be more efficient promoter of IL-10 production as well as weaker inducer of NO production and secretion of pro-inflammatory cytokines TNFα, and IL-6. The ability of rBet v 1-BanLec to promote IL-10 in together with the preserved 3D structure of Bet v 1 part implies that the construct might exert a beneficial effect in the allergen-specific immunotherapy.

过敏原特异性免疫疗法 (AIT) 是一种针对过敏性疾病的脱敏疗法，可纠正对无害蛋白质抗原（称为过敏原）的强调病理性免疫反应。AIT 中使用的重组过敏原允许生产定义明确的配方，这些配方具有一致的质量，但效率通常低于天然过敏原提取物。将重组过敏原与佐剂或免疫调节剂结合可以提高 AIT 的功效。本研究旨在对新设计的由主要桦树花粉过敏原 Bet v 1 和香蕉凝集素 (BanLec)、TLR2 和 CD14 结合蛋白组成的重组嵌合体进行结构和功能表征，以用于 AIT。rBet v 1-BanLec 嵌合体是用计算机设计的并在大肠杆菌中表达为可溶性组分. 纯化的 rBet v 1-BanLec (33.4 kDa) 保留了与 BanLec 相关的碳水化合物结合生物学活性，并保留了 Bet v 1 的 IgE 反应性表位。嵌合体显示出具有主要 β 折叠的二级结构。与 rBanLec 和 rBet v 1 相比，在巨噬细胞上测试的 rBet v 1-BanLec 的免疫调节能力显示髓过氧化物酶活性发生变化，NO 产生减少，细胞因子产生显着改变。 与 rBet v 1 相比，rBet v 1- BanLec 被证明是更有效的 IL-10 产生促进剂，以及较弱的 NO 产生和促炎细胞因子 TNFα 和 IL-6 分泌的诱导剂。