Centro de Biotecnología y Genómica de Plantas (CBGP), Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), Universidad Politécnica de Madrid (UPM), Madrid, Spain.
Departamento de Biotecnología-Biología Vegetal, Escuela Técnica Superior de Ingeniería Agronómica, Alimentaria y de Biosistemas, Universidad Politécnica de Madrid (UPM), Madrid, Spain.
Sci Rep. 2022 Feb 28;12(1):3329. doi: 10.1038/s41598-022-07421-y.
Allergic sensitization is initiated by protein and epithelia interaction, although the molecular mechanisms leading this encounter toward an allergic phenotype remain unknown. Here, we apply the two-hit hypothesis of inflammatory diseases to the study of food allergy sensitization. First, we studied the effects of long-term depilation in mice by analyzing samples at different time points. Several weeks of depilation were needed until clear immunological changes were evidenced, starting with upregulation of NLRP3 protein levels, which was followed by overexpression of Il1b and Il18 transcripts. Secondly, we assessed the effects of allergen addition (in this case, Pru p 3 in complex with its natural lipid ligand) over depilated skin. Systemic sensitization was evaluated by intraperitoneal provocation with Pru p 3 and measure of body temperature. Anaphylaxis was achieved, but only in mice sensitized with Prup3_complex and not treated with the NLRP3 inhibitor MCC950, thus demonstrating the importance of both hits (depilation + allergen addition) in the consecution of the allergic phenotype. In addition, allergen encounter (but not depilation) promoted skin remodeling, as well as CD45+ infiltration not only in the sensitized area (the skin), but across several mucosal tissues (skin, lungs, and gut), furtherly validating the systemization of the response. Finally, a low-scale study with human ILC2s is reported, where we demonstrate that Prup3_complex can induce their phenotype switch (↑CD86, ↑S1P1) when cultured in vitro, although more data is needed to understand the implications of these changes in food allergy development.
过敏敏化是由蛋白质和上皮细胞相互作用引发的，尽管导致这种过敏表型的分子机制尚不清楚。在这里，我们将炎症性疾病的双打击假说应用于食物过敏敏化的研究。首先，我们通过分析不同时间点的样本研究了长期脱毛对小鼠的影响。需要数周的脱毛时间才能出现明显的免疫变化，首先是 NLRP3 蛋白水平上调，随后 Il1b 和 Il18 转录本过表达。其次，我们评估了过敏原添加（在这种情况下，Pru p 3 与天然脂质配体复合物）对脱毛皮肤的影响。通过腹腔内用 Pru p 3 激发和测量体温来评估全身致敏。仅在用 Prup3_complex 致敏且未用 NLRP3 抑制剂 MCC950 处理的小鼠中实现了过敏反应，但仅在脱毛和过敏原添加（depilation + allergen addition）的双打击后才能实现过敏表型。此外，过敏原接触（而非脱毛）促进了皮肤重塑，以及不仅在致敏区域（皮肤），而且在几个黏膜组织（皮肤、肺和肠道）中 CD45+浸润，进一步验证了该系统的反应。最后，报告了一项人类 ILC2 的小规模研究，其中我们证明了当在体外培养时，Prup3_complex 可以诱导其表型转换（↑CD86，↑S1P1），尽管需要更多的数据来了解这些变化对食物过敏发展的影响。
