Department of Food, Environmental and Nutritional Sciences (DeFENS), University of Milan, Via Celoria 2, 20133 Milan, Italy.
Department of Veterinary and Animal Sciences, University of Copenhagen, Ridebanevej 9, 1871 Frederiksberg, Denmark.
Biomolecules. 2023 Oct 16;13(10):1531. doi: 10.3390/biom13101531.
γ-Conglutin (γ-C) from lupin seeds has been identified as a potent allergen with cross reactivity to peanuts. Here, we investigated how γ-C affected the response in bone marrow-derived dendritic cells (DCs) to bacterial stimuli. γ-C enhanced NCFM (LaNCFM)-induced IL-12, IL-10, and IL-23 dose-dependently. In contrast, together with Nissle or LPS, γ-C reduced the production of IL-12 but not of IL-23 and IL-10. Enzyme-hydrolyzed γ-C also enhanced LaNCFM-induced IL-12 and IL-23 production. All preparations induced ROS production in the DCs. The mannose receptor ligands mannan and dextran and the clathrin inhibitor monodansylcadaverine partly inhibited the endocytosis of γ-C. Kunitz trypsin inhibitor and the scavenger receptor ligand polyG also enhanced LaNCFM-induced IL-12, indicating the involvement of receptors other than C-type lectin receptors. The endocytosis of labeled γ-C increased dose-dependently by addition of unlabeled γ-C, which coincided with γ-C's tendency to aggregate. Taken together, γ-C aggregation affects endocytosis and affects the cytokine production induced by gram-positive and gram-negative bacteria differently. We suggest that γ-C is taken up by the same mechanism as other food proteins but due to aggregation is present in higher concentration in the DCs. This could influence the resulting T-cell response in a microbial stimuli-dependent way.
γ-羽扇豆球蛋白（γ-C）已被鉴定为一种具有与花生交叉反应性的强过敏原。在这里，我们研究了 γ-C 如何影响骨髓来源树突状细胞（DC）对细菌刺激的反应。γ-C 增强了 NCFM（LaNCFM）诱导的 IL-12、IL-10 和 IL-23 的产生呈剂量依赖性。相比之下，与 Nissle 或 LPS 一起，γ-C 减少了 IL-12 的产生，但不减少 IL-23 和 IL-10 的产生。酶水解的 γ-C 也增强了 LaNCFM 诱导的 IL-12 和 IL-23 的产生。所有制剂均在 DC 中诱导 ROS 产生。甘露糖受体配体甘露聚糖和葡聚糖以及网格蛋白抑制剂单丹磺酰尸胺部分抑制了 γ-C 的内吞作用。Kunitz 胰蛋白酶抑制剂和清道夫受体配体聚鸟氨酸也增强了 LaNCFM 诱导的 IL-12，表明除 C 型凝集素受体外，还涉及其他受体。随着未标记的 γ-C 的添加，标记的 γ-C 的内吞作用呈剂量依赖性增加，这与 γ-C 聚集的趋势一致。总之，γ-C 的聚集会影响内吞作用，并以不同的方式影响革兰氏阳性和革兰氏阴性细菌诱导的细胞因子产生。我们建议 γ-C 通过与其他食物蛋白相同的机制被摄取，但由于聚集作用，在 DC 中以更高的浓度存在。这可能会以依赖微生物刺激的方式影响随后的 T 细胞反应。
