Moreno F Javier, Rubio Luis A, Olano Agustín, Clemente Alfonso
Instituto de Fermentaciones Industriales (CSIC), C/Juan de la Cierva 3, 28006 Madrid, Spain.
J Agric Food Chem. 2006 Nov 1;54(22):8631-9. doi: 10.1021/jf061760h.
We have investigated the absorption rates of two purified major allergen 2S albumins, Ber e 1 from Brazil nuts (Bertholletia excelsa Humb. & Bonpl.) and Ses i 1 from white sesame seeds (Sesamum indicum L.), across human intestinal epithelial Caco-2 cell monolayers following gastrointestinal digestion in vitro. The transport from apical to basolateral side in cell monolayers was evaluated by RP-HPLC-UV and indirect competitive ELISA methods, being confirmed by western-blotting analysis. Significant amounts (approximately 15-25 nmol micromol(-1) initial amount/h) of intact Ber e 1 and Ses i 1 were found in the basolateral side. The absorption rates of both plant allergens through the cell monolayer were shown to be constant during the whole incubation period (4 h at 37 degrees C), verifying that the permeability of the membrane was not altered by the allergen digests. Our findings revealed that both purified 2S albumin allergens may be able to survive in immunologically reactive forms to the simulated harsh conditions of the gastrointestinal tract to be transported across the Caco-2 cell monolayers, so that they would be able to sensitize the mucosal immune system and/or elicit an allergic response.
我们研究了两种纯化的主要过敏原2S白蛋白,即来自巴西坚果(Bertholletia excelsa Humb. & Bonpl.)的Ber e 1和来自白芝麻籽(Sesamum indicum L.)的Ses i 1,在体外经胃肠道消化后通过人肠上皮Caco-2细胞单层的吸收速率。通过反相高效液相色谱-紫外检测法(RP-HPLC-UV)和间接竞争酶联免疫吸附测定法(ELISA)评估细胞单层中从顶端到基底外侧的转运,并通过蛋白质印迹分析进行了确认。在基底外侧发现了大量完整的Ber e 1和Ses i 1(约15 - 25 nmol/μmol初始量/小时)。两种植物过敏原通过细胞单层的吸收速率在整个孵育期(37℃下4小时)内保持恒定,证实了膜的通透性不会因过敏原消化物而改变。我们的研究结果表明,两种纯化的2S白蛋白过敏原可能能够以免疫反应性形式在模拟的胃肠道恶劣条件下存活,从而穿过Caco-2细胞单层进行转运,进而能够使黏膜免疫系统致敏和/或引发过敏反应。