Peeking into the Stingers: A Comprehensive SWATH-MS Study of the European Hornet Vespa crabro (Linnaeus, 1758) (Hymenoptera: Vespidae) Venom Sac Extracts

This study aimed to investigate the venom sac extracts (VSE) of the European hornet (EH) Vespa crabro (Linnaeus, 1758) (Hymenoptera: Vespidae), focusing on the differences between stinging females, gynes (G) and workers (W), at the protein level. Using a quantitative “Sequential Window Acquisition of all Theoretical Fragment Ion Mass Spectra” (SWATH-MS) analysis, we identified and quantified a total of 240 proteins. Notably, within the group, 45.8 % (n = 110) showed significant differential expression between VSE-G and VSE-W. In this set, 57.3 % (n = 63) were upregulated and 42.7 % (n = 47) downregulated in the G. Additionally, the 200 quantified proteins from the class Insecta belong to 16 different species, six of them to he Hymenoptera/Apidae lineage, comprising seven proteins with known potential as allergens. Phospholipase A1 (Vesp v 1), phospholipase A1 verutoxin 2b (VT-2b), hyaluronidase A (Vesp v 2A), hyaluronidase B (Vesp v 2B), and venom allergen 5 (Vesp v 5) were significantly downregulated in the G, and vitellogenin (Ves v 6) was upregulated. Overall, 46 % of the VSE proteins showed differential expression, with a majority being upregulated in G. These findings shed light on the proteomic differences in VSE between EH castes, potentially contributing to our understanding of their behavior and offering insights for allergy research.

本研究旨在探究欧洲胡蜂（European hornet, EH）Vespa crabro（Linnaeus, 1758）（膜翅目：胡蜂科）的毒囊提取物（venom sac extracts, VSE），并从蛋白质组层面聚焦螫刺雌性蜂、处女蜂王（gynes, G）与工蜂（workers, W）之间的差异。本研究采用定量分析技术「全理论碎片离子质谱连续窗口采集」（Sequential Window Acquisition of all Theoretical Fragment Ion Mass Spectra, SWATH-MS），共鉴定并定量得到240种蛋白质。值得注意的是，该群体中45.8%（n=110）的蛋白质在VSE-G与VSE-W间存在显著差异表达。其中57.3%（n=63）的差异蛋白在G组中呈上调趋势，42.7%（n=47）呈下调趋势。此外，本次定量得到的200种昆虫纲（Insecta）蛋白质分属16个不同物种，其中6个物种属于膜翅目/蜜蜂科（Hymenoptera/Apidae）谱系，包含7种已被证实具有潜在致敏原活性的蛋白质。磷脂酶A1（Vesp v 1）、磷脂酶A1毒毒素2b（phospholipase A1 verutoxin 2b, VT-2b）、透明质酸酶A（Vesp v 2A）、透明质酸酶B（Vesp v 2B）以及毒液变应原5（Vesp v 5）在G组中显著下调；而卵黄蛋白原（Ves v 6）则呈上调趋势。总体而言，46%的VSE蛋白质存在差异表达，且其中大多数在G组中呈上调状态。本研究结果揭示了欧洲胡蜂不同社会品级毒囊提取物的蛋白质组学差异，或有助于深化对其行为模式的理解，并为变应原研究提供新的视角。