IgE cross-inhibition between Ara h 1 and Ara h 2 is explained by complex formation of both major peanut allergens.
IgE cross-inhibition between Ara h 1 and Ara h 2 is explained by complex formation of both major peanut allergens.
复制标题
DOI：
10.1016/j.jaci.2023.03.025
复制
发表时间：
2023-08
影响因子：
14.2
通讯作者：
Ronald van Ree
中科院分区：
文献类型：
--
作者：
Hans J.M. Warmenhoven;Luuk Hulsbos;Stephen C. Dreskin;Jaap H. Akkerdaas;Serge A. Versteeg;Ronald van Ree
令人惊讶的是，尽管序列同源性很低，但报道了主要花生过敏原Arah1、2和3之间的IgE交叉反应。我们调查了花生主要过敏原之间意想不到的交叉反应。用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳法(SDS-PAGE)、Western印迹试验、LC-MS/MS和夹心酶联免疫吸附试验(EL ISA)评价纯化的天然阿糖胞苷1、2、3和6的交叉污染情况。用完整的天然变应原、重组变应原和代表Arah1和Arah2交叉反应表位的合成肽，用ELISA法和免疫CAP抑制法研究了花生过敏患者(n=43)的IgE交叉反应。经夹心ELISA法、SDS-PAGE/Western印迹分析和LC-MS/MS分析表明，纯化的Narah1和Narah3都含有少量但相当数量的Arah2和Arah6(<；1%)，2S白蛋白与Arah1和Arah3之间的IgE交叉抑制仅在使用天然纯化变应原时观察到，而不是重组变应原或合成肽。当纯化的Narah1在还原条件下处理时，失去了明显的交叉反应，这表明Arah2和Arah6污染物可能通过二硫键相互作用共价结合到arah1上。花生2S蛋白与Arah1和Arah3不能发生真正的交叉反应。相反，少量的交叉污染被证明足以引起显著的交叉抑制，这可能被误解为分子交叉反应。使用纯化的Narah1和Narah3的诊断试验可能会高估它们作为主要过敏原的重要性，因为存在污染的2S白蛋白，使重组arah1和arah3成为首选替代方案。
Surprisingly, IgE cross-reactivity between the major peanut allergens Ara h 1, 2, and 3 has been reported despite very low sequence identities. We investigated the unexpected cross-reactivity between peanut major allergens. Cross-contamination of purified natural Ara h 1, 2, 3, and 6 was assessed by sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE), Western blot test, liquid chromatography–tandem mass spectrometry (LC-MS/MS), and sandwich enzyme-linked immunosorbent assay (ELISA). IgE cross-reactivity was studied with sera of peanut-allergic patients (n = 43) by ELISA and ImmunoCAP inhibition using both intact natural and recombinant allergens and synthetic peptides representing postulated Ara h 1 and Ara h 2 cross-reactive epitopes. Both purified nAra h 1 and nAra h 3 were demonstrated to contain small but significant amounts of Ara h 2 and Ara h 6 (<1%) by sandwich ELISA, SDS-PAGE/Western blot analysis, and LC-MS/MS. IgE cross-inhibition between both 2S albumins and Ara h 1 and Ara h 3 was only observed when using natural purified allergens, not recombinant allergens or synthetic peptides. Apparent cross-reactivity was lost when purified nAra h 1 was pretreated under reducing conditions, suggesting that Ara h 2 and Ara h 6 contaminations may be covalently bound to Ara h 1 via disulfide interactions. True cross-reactivity of both peanut 2S albumins with Ara h 1 and Ara h 3 could not be demonstrated. Instead, cross-contamination with small quantities was shown to be sufficient to cause significant cross-inhibition that can be misinterpreted as molecular cross-reactivity. Diagnostic tests using purified nAra h 1 and nAra h 3 can overestimate their importance as major allergens as a result of the presence of contaminating 2S albumins, making recombinant Ara h 1 and Ara h 3 a preferred alternative.
影响因子：
2.8
作者：
Chen X;Wang Q;El-Mezayen R;Zhuang Y;Dreskin SC
通讯作者：
Dreskin SC