Gal d 7-a major allergen in primary chicken meat allergy
Gal d 7-a major allergen in primary chicken meat allergy
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DOI：
10.1016/j.jaci.2020.02.033
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发表时间：
2020-07-01
影响因子：
14.2
通讯作者：
Swoboda, Ines
中科院分区：
文献类型：
--
作者：
Klug, Christoph;Hemmer, Wolfgang;Swoboda, Ines
背景：禽肉可引起严重的过敏反应。到目前为止，导致禽肉过敏的分子在很大程度上是未知的。目的：鉴定和表征禽肉过敏原。方法：采用免疫印迹法分析患者对鸡肌肉的IgE反应谱，并对大多数患者识别的蛋白进行肽质量指纹图谱分析。鉴定出一个23 kda的ige反应蛋白为肌球蛋白轻链1，命名为Gallus domesticus 7 （Gal d 7）。重组Gal d 7在大肠杆菌中得到。ELISA实验分析该蛋白的IgE反应性，抑制免疫印迹法评估其与其他禽类过敏原的交叉反应性。采用圆二色性分析评价其折叠稳定性和热稳定性，采用体外胃肠消化试验考察其酶促稳定性。结果：重组gald7是一种正确折叠的，主要是α -螺旋蛋白，并显示出与天然对应物相当的ige结合活性。对28例鸡肉过敏患者的IgE反应性分析表明，gald7是主要对鸡肉敏感的患者的主要过敏原。此外，在其他禽类中也检测到Gal d 7交叉反应过敏原，表明重组Gal d 7可以作为家禽肉类过敏的诊断标记过敏原。高热稳定性、再折叠能力和对胃肠道酶的抗性可能解释了为什么Gal d 7可以作为一种有效的增敏剂。结论：gald7是一种新型的主要鸡肉过敏原。重组gald7可用于家禽肉致敏诊断。
Background: Poultry meat can induce severe allergic reactions. So far, the molecules causing poultry meat allergy are largely unknown.Objective: Our aim was to identify and characterize poultry meat allergens.Methods: Profiles of patients' IgE reactivity to chicken muscle were analyzed in immunoblots, and proteins recognized by the majority of patients were subjected to peptide mass fingerprinting. A 23-kDa IgE-reactive protein was identified as myosin light chain 1, designated Gallus domesticus 7 (Gal d 7). Recombinant Gal d 7 was produced in Escherichia coli. The protein's IgE reactivity was analyzed in ELISA experiments, and cross-reactivity with allergens of other poultry species was assessed in inhibition immunoblots. Fold and thermal stability were evaluated by circular dichroism analysis, and enzymatic stability was investigated using in vitro gastrointestinal digestion assays.Results: Recombinant Gal d 7 represents a properly folded, predominantly alpha-helical protein and displays IgE-binding activity comparable to that of its natural counterpart. IgE reactivity analysis in 28 patients allergic to chicken meat revealed that Gal d 7 is a major allergen for patients primarily sensitized to chicken meat. Furthermore, Gal d 7-cross-reactive allergens were also detected in other poultry species, suggesting that recombinant Gal d 7 can be used as a diagnostic marker allergen for poultry meat allergy. The high thermal stability, refolding capacity, and resistance to gastrointestinal enzymes might explain why Gal d 7 can act as a potent sensitizing agent.Conclusion: Gal d 7 represents a novel major chicken meat allergen. Recombinant Gal d 7 could be used for diagnosis of genuine poultry meat sensitization.