The Netherlands Organisation for Applied Scientific Research TNO, Princetonlaan 6, Utrecht, Utrecht, 3584 CB, The Netherlands.
Department of Dermatology/Allergology, University Medical Center Utrecht, Utrecht University, Heidelberglaan 100, Utrecht, Utrecht, 3584 CX, The Netherlands.
Mol Nutr Food Res. 2021 Mar;65(6):e2000712. doi: 10.1002/mnfr.202000712. Epub 2021 Jan 25.
No accepted and validated methods are currently available which can accurately predict protein allergenicity. In this study, the role of digestion and transport on protein allergenicity is investigated.
Peanut allergens (Ara h 1, 2, 3, and 6) and a milk allergen (β-lactoglobulin) are transported across pig intestinal epithelium using the InTESTine model and afterward basophil activation is measured to assess the (remaining) functional properties. Additionally, allergens are digested by pepsin prior to epithelial transport and their allergenicity is assessed in a human mast cell activation assay. Remarkably, transported Ara h 1 and 3 are not able to activate basophils, in contrast to Ara h 2 and 6. Digestion prior to transport results in a significant increase in mast cell activation of Ara h 1 and 3 dependent on the length of digestion time. Activation of mast cells by Ara h 2 and 6 is unaffected by digestion prior to transport.
Digestion and transport influences the allergenicity of Ara h 1 and 3, but not of Ara h 2 and 6. The influence of digestion and transport on protein allergenicity may explain why current in vitro assays are not predictive for allergenicity.
目前尚无可准确预测蛋白质变应原性的公认和验证方法。本研究探讨了消化和转运对蛋白质变应原性的作用。
使用 InTESTine 模型转运花生过敏原（Ara h 1、2、3 和 6）和牛奶过敏原（β-乳球蛋白）穿过猪肠上皮细胞，然后测量嗜碱性粒细胞活化以评估（剩余）功能特性。此外，在肠上皮细胞转运之前，用胃蛋白酶消化过敏原，并在人肥大细胞活化测定中评估其变应原性。值得注意的是，与 Ara h 2 和 6 相比，转运的 Ara h 1 和 3 不能激活嗜碱性粒细胞。转运前的消化导致 Ara h 1 和 3 的肥大细胞活化显著增加，这取决于消化时间的长短。转运前的消化对 Ara h 2 和 6 诱导肥大细胞活化没有影响。
消化和转运影响 Ara h 1 和 3 的变应原性，但不影响 Ara h 2 和 6 的变应原性。消化和转运对蛋白质变应原性的影响可能解释了为什么目前的体外检测方法不能预测变应原性。
