Transglutaminase-mediated cross-linking of a peptic fraction of omega-5 gliadin enhances IgE reactivity in wheat-dependent, exercise-induced anaphylaxis.
Transglutaminase-mediated cross-linking of a peptic fraction of omega-5 gliadin enhances IgE reactivity in wheat-dependent, exercise-induced anaphylaxis.
复制标题
转谷氨酰胺酶介导的 omega-5 麦醇溶蛋白消化部分的交联可增强小麦依赖性、运动诱发的过敏反应中的 IgE 反应性。
DOI：
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发表时间：
2003
影响因子：
14.2
通讯作者：
H. Alenius
中科院分区：
文献类型：
--
作者：
K. Palosuo;E. Varjonen;J. Nurkkala;N. Kalkkinen;R. Harvima;T. Reunala;H. Alenius
背景
患有小麦依赖性运动诱发过敏反应（WDEIA）的患者在摄入小麦制品后运动时会出现反复过敏反应。我们已经确定ω-5麦醇溶蛋白（Tri a 19）是WDEIA的主要过敏原，但运动在引发症状中的作用仍然不清楚。
目的
目的是检查组织转氨酶（tTG）介导的交联是否可以参与调节IgE结合能力和消化的ω-5麦醇溶蛋白肽在WDEIA中的体内反应性。
方法
用胃蛋白酶或用胃蛋白酶和胰蛋白酶消化纯化的ω-5麦醇溶蛋白并用tTG处理。在消化肽的tTG处理之前和之后，通过免疫印迹法研究了来自10名WDEIA患者的合并血清中IgE抗体的结合。通过凝胶过滤色谱法分离来自胃蛋白酶消化的肽，并通过免疫印迹法研究tTG处理前后4种不同肽级分的IgE反应性。通过IgE ELISA、ELISA抑制和皮肤点刺试验进一步研究了通过tTG显示最大程度交联的级分。
结果
在胃蛋白酶和胃蛋白酶-胰蛋白酶消化后，ω-5麦醇溶蛋白的IgE结合能力得以保留。tTG处理的整个消化物形成大的肽复合物，分子量范围从40到大于200 kd。这些交联的聚集体在免疫印迹中比未处理的、胃蛋白酶消化的或胃蛋白酶-胰蛋白酶消化的ω-5麦醇溶蛋白更强烈地结合IgE抗体。对应于色谱图的最高峰并显示最大程度的tTG介导的交联的整个消化酶消化物的凝胶过滤级分显示在tTG处理后ELISA中血清IgE反应性增加，以及对交联复合物的反应性转移。在20例WDEIA患者中，由这种经tTG处理的消化性组分引起的平均皮肤点刺试验风团比未经处理的消化性组分引起的大77%（P <0.001），比完整的ω-5麦胶蛋白引起的大56%（P <0.01）。
结论
ω-5麦胶蛋白衍生肽通过tTG交联，这导致体外和体内IgE结合显著增加。WDEIA患者运动期间肠粘膜中tTG的激活可能导致能够引发过敏反应的大变应原复合物的形成。
BACKGROUND
Patients with wheat-dependent, exercise-induced anaphylaxis (WDEIA) experience recurrent anaphylactic reactions when exercising after ingestion of wheat products. We have identified omega-5 gliadin (Tri a 19) as a major allergen in WDEIA, but the role of exercise in eliciting the symptoms remains obscure.
OBJECTIVE
The aim was to examine whether tissue transglutaminase (tTG)-mediated cross-linking could be involved in modulating the IgE-binding ability and in vivo reactivity of digested omega-5 gliadin peptides in WDEIA.
METHODS
Purified omega-5 gliadin was digested with pepsin or with pepsin and trypsin and treated with tTG. The binding of IgE antibodies in pooled sera from 10 patients with WDEIA was studied by means of immunoblotting before and after tTG treatment of the digested peptides. The peptides derived from pepsin digestion were separated by means of gel-filtration chromatography, and IgE reactivity of 4 different peptide fractions was studied by immunoblotting before and after tTG treatment. The fraction showing the greatest degree of cross-linking by tTG was further studied by means of IgE ELISA, ELISA inhibition, and skin prick testing.
RESULTS
The IgE-binding ability of omega-5 gliadin was retained after pepsin and pepsin-trypsin digestion. tTG treatment of the whole peptic digest formed large peptide complexes, with molecular weights ranging from 40 to greater than 200 kd. These cross-linked aggregates bound IgE antibodies in immunoblotting more intensely than untreated, pepsin-digested, or pepsin-trypsin-digested omega-5 gliadin. A gel-filtration fraction of the whole peptic digest corresponding to the highest peak of the chromatogram and showing the greatest degree of tTG-mediated cross-linking showed an increase in serum IgE reactivity in ELISA after tTG treatment, as well as a shift of reactivity to cross-linked complexes. In the 20 patients with WDEIA, the mean skin prick test wheal elicited by this tTG-treated peptic fraction was 77% larger (P <.001) than that elicited by the untreated peptic fraction and 56% larger (P <.01) than that elicited by intact omega-5 gliadin.
CONCLUSIONS
Omega-5 gliadin-derived peptides are cross-linked by tTG, which causes a marked increase in IgE binding both in vitro and in vivo. Activation of tTG during exercise in the intestinal mucosa of patients with WDEIA could lead to the formation of large allergen complexes capable of eliciting anaphylactic reactions.
DOI：
10.1152/ajpgi.1997.272.2.g281
发表时间：
1997-02-01
影响因子：
4.5
作者：
Mirza, A;Liu, SL;Zern, MA
通讯作者：
Zern, MA
影响因子：
3.3
作者：
Pals, KL;Chang, RT;Gisolfi, CV
通讯作者：
Gisolfi, CV