Laboratory of Immunogenetics and Allergology, CRP-Santé, L-1526 Luxembourg, Luxembourg.
Clin Exp Allergy. 2013 Jul;43(7):811-22. doi: 10.1111/cea.12117.
The majority of fish-allergic patients are sensitized to parvalbumin, known to be the cause of important IgE cross-reactivity among fish species. Little is known about the importance of fish allergens other than parvalbumin.
The aim of this study was to characterize hitherto undefined fish allergens in three commonly consumed fish species, cod, salmon and tuna, and to evaluate their importance for in vitro IgE-diagnosis in addition to parvalbumin and fish gelatin.
Sixty-two patients were diagnosed by clinical history, skin prick tests and specific IgE to fish extracts. Two new fish allergens from cod, salmon and tuna were identified by microsequencing. These proteins were characterized by immunoblot, ELISA and mediator release assay. Purified parvalbumin, enolase, aldolase and fish gelatin were used for quantification of specific IgE in ELISA.
Parvalbumin and two other allergens of 50 and 40 kDa were detected in IgE-immunoblots of cod, salmon and tuna extracts by most patient sera. The 50 and 40 kDa proteins were identified as beta-enolase and fructose-bisphosphate aldolase A respectively. Both purified enzymes showed allergenic activity in the mediator release assay. Indeed, 72.6% of the patients were sensitized to parvalbumin, 20% of these had specific IgE to salmon parvalbumin only. IgE to enolases were found in 62.9% (0.5-95.0 kUA /L), to aldolases in 50.0% (0.4-26.0 kUA /L) and to fish gelatin in 19.3% (0.4-20.0 kUA /L) of the patients. Inter-species cross-reactivity, even though limited, was found for enolases and aldolases by IgE-inhibition ELISA.
Fish enolase and aldolase have been identified as important new fish allergens. In fish allergy diagnosis, IgE to enolase and aldolase are especially relevant when IgE to parvalbumin are absent.
大多数鱼类过敏患者对副肌球蛋白敏感，已知副肌球蛋白是鱼类之间重要 IgE 交叉反应的原因。然而，除了副肌球蛋白和鱼明胶之外，对其他鱼类过敏原的了解甚少。
本研究的目的是鉴定三种常见食用鱼类（鳕鱼、三文鱼和金枪鱼）中迄今尚未定义的鱼类过敏原，并评估它们除副肌球蛋白和鱼明胶之外，对体外 IgE 诊断的重要性。
62 例患者通过临床病史、皮肤点刺试验和鱼提取物特异性 IgE 进行诊断。通过微测序鉴定了来自鳕鱼、三文鱼和金枪鱼的两种新的鱼类过敏原。通过免疫印迹、ELISA 和介质释放测定法对这些蛋白质进行了特征描述。使用纯化的副肌球蛋白、烯醇酶、醛缩酶和鱼明胶进行 ELISA 中特异性 IgE 的定量。
通过大多数患者血清的鳕鱼、三文鱼和金枪鱼提取物的 IgE-免疫印迹检测到副肌球蛋白和另外两种 50 和 40 kDa 的过敏原。50 和 40 kDa 蛋白分别被鉴定为β-烯醇酶和果糖-1,6-二磷酸醛缩酶 A。两种纯化的酶在介质释放测定中均显示出过敏原活性。事实上，72.6%的患者对副肌球蛋白过敏，其中 20%的患者仅对三文鱼副肌球蛋白有特异性 IgE。在 62.9%（0.5-95.0 kUA /L）的患者中发现了对烯醇酶的 IgE，在 50.0%（0.4-26.0 kUA /L）的患者中发现了对醛缩酶的 IgE，在 19.3%（0.4-20.0 kUA /L）的患者中发现了对鱼明胶的 IgE。通过 IgE 抑制 ELISA 发现烯醇酶和醛缩酶之间存在有限的种间交叉反应。
已鉴定出鱼类烯醇酶和醛缩酶为重要的新鱼类过敏原。在鱼类过敏诊断中，当 IgE 对副肌球蛋白缺失时，IgE 对烯醇酶和醛缩酶尤其相关。
