Cell-based immunological assay: complementary applications in evaluating the allergenicity of foods with FAO/WHO guidelines
Cell-based immunological assay: complementary applications in evaluating the allergenicity of foods with FAO/WHO guidelines
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基于细胞的免疫学检测：根据FAO/WHO指南评估食品过敏性的补充应用
DOI：
10.1016/j.foodres.2014.04.033
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发表时间：
2014-08-01
影响因子：
8.1
通讯作者：
Che, Huilian
中科院分区：
文献类型：
--
作者：
Sun, Na;Wang, Jing;Che, Huilian
粮食及农业组织（粮农组织）和世界卫生组织（卫生组织）（粮农组织/卫生组织，2001年）建议的决策树战略在致敏性评估方面存在缺陷。在本研究中，我们采用RBL细胞免疫试验和决策树策略评估了甘氨酸（Gly）、卵清蛋白（OVA）和马铃薯酸性磷酸酶（PAP）的致敏活性。然后，我们进一步分析了基于RBL细胞的免疫测定和决策树策略之间的互补应用。生物信息学分析预测潜在致敏性排序为Gly b> OVA > PAP，通过分析线性表位区域确定。然而，基于RBL细胞的免疫分析可以确定线性和构象表位诱导过敏反应的能力。胃蛋白酶消化试验表明，Gly比OVA更稳定，而PAP在模拟胃液中没有稳定性。然而，这种方法不能完全反映食物蛋白质在胃肠道消化的结果。Gly加OVA诱导Balb/c小鼠的过敏反应及RBL-2H3细胞β -己糖氨酸酶的最佳释放；PAP无显著反应，释放量< 10%。与动物模型相比，基于RBL细胞的免疫试验基于检测系统的简单特性，更适合于评价食品蛋白质的致敏活性。当生物信息学分析和胃蛋白酶消化分析结果不一致时，基于RBL细胞的免疫试验可以发挥“裁判”作用，动物模型可以用于进一步剖析过敏反应的机制。因此，基于RBL细胞的免疫分析与决策树策略之间的互补应用将是对食品致敏潜力进行全面评估的良好选择。(C) 2014 Elsevier Ltd.版权所有。
There are drawbacks for the decision tree strategies recommended by the Food and Agriculture Organization (FAO) and the World Health Organization (WHO) (FAO/WHO, 2001) in allergenicity assessment. In this study, we evaluated the allergenic activity of glycinin (Gly), ovalbumin (OVA) and potato acid phosphatase (PAP) by using an RBL cell-based immunological assay and the decision tree strategies. We then further analyzed the complementary applications between the RBL cell-based immunological assay and the decision tree strategies. A bioinformatical analysis predicted that the ranking of potential allergenicity was Gly > OVA > PAP, which was determined by analyzing the linear-epitope regions. However, the RBL cell-based immunological assay can determine the ability of the linear and conformational epitopes to induce allergic reactions. A pepsin digestion assay indicated that Gly was more stable compared with OVA and that PAP had no stability in simulated gastric fluid. However, this approach cannot fully reflect the results of food proteins digested in the gastrointestinal tract. Anaphylactic reactions in Balb/c mice and optimal beta-hexosaminidase release of RBL-2H3 cells were induced with Gly followed by OVA; no significant response and < 10% release were obtained with PAP. Compared with the animal models, the RBL cell-based immunological assay is more suitable for evaluating the allergenic activity of food proteins based on the simple characteristics of the testing system. The RBL cell-based immunological assay may play a "referee" role when inconsistent results between a bioinformatical analysis and a pepsin digestion assay occur, and animal models may be used to further dissect the mechanisms of the allergic responses. Thus complementary applications between the RBL cell-based immunological assay and the decision tree strategies will be a good choice to conduct a complete evaluation of the allergenic potential of foods. (C) 2014 Elsevier Ltd. All rights reserved.