中华皮肤科杂志 ›› 2015, Vol. 48 ›› Issue (6): 369-372.

• 论著 •    下一篇

诱发儿童特应性皮炎的三疣梭子蟹过敏原研究

路晓琳1,陈官芝2,王国英3,李振兴4,苏磊1,王宗岭1   

  1. 1. 青岛大学附属医院
    2. 青岛大学医学院附属医院
    3. 青岛大学医学院 免疫学教研室
    4. 中国海洋大学食品学院
  • 收稿日期:2015-01-15 修回日期:2015-01-29 出版日期:2015-06-15 发布日期:2015-06-03
  • 通讯作者: 陈官芝 E-mail:chenguanzhi@outlook.com
  • 基金资助:

    肾病综合征及免疫性疾病的诊断与治疗的研究-儿童特应性皮炎食物过敏原的筛查与分子结构分析

Identification of allergens in portunus trituberculatus responsible for atopic dermatitis in children

  • Received:2015-01-15 Revised:2015-01-29 Online:2015-06-15 Published:2015-06-03

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摘要：

目的 探讨诱发儿童特应性皮炎的三疣梭子蟹过敏原。 方法 对入选的儿童特应性皮炎患儿进行血清蟹过敏原特异性IgE抗体检测或蟹过敏原皮肤点刺试验，阳性者进行三疣梭子蟹口服食物激发试验，选择出对蟹过敏的患儿33例，留取血清作为实验组；选择同期健康体检儿童30例，留取血清作为对照组。选择新鲜的三疣梭子蟹，提取总蛋白，用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）、免疫印迹检测诱发儿童特应性皮炎的蟹过敏原。 结果 三疣梭子蟹蛋白粗提液可见条带有11条，相对分子质量分别为94 000、70 000、58 000、49 000、36 000、34 000、32 000、27 000、21 000、19 000、17 000。能引起实验组免疫反应的条带有4条，相对分子质量分别为70 000、58 000、49 000、36 000，实验组阳性率分别为93.9%、45.4%、39.4%、100%；对照组无阳性条带。将实验组相对分子质量70 000、58 000、49 000及36 000免疫条带阳性率分别与健康对照组比较，χ2分别为55.483、17.898、14.891、63.000，均P < 0.05。 结论 70 000、36 000蛋白是诱发儿童特应性皮炎的三疣梭子蟹主要过敏原。

Abstract:

Lu Xiaolin*, Chen Guanzhi, Wang Guoying, Li Zhenxing, Su Lei, Wang Zongling. *Department of Dermatology, Affiliated Hospital of Qingdao University, Qingdao 266003, China Corresponding author: Chen Guanzhi, Email: chenguanzhi@outlook.com 【Abstract】 Objective To identify allergens in portunus trituberculatus responsible for atopic dermatitis （AD） in children. Methods Totally, 145 child outpatients with AD were enrolled in this study from September 2013 to July 2014, and underwent the skin prick test （SPT） with crab proteins or crab-specific IgE determination assay. Then, the children with positive SPT or elevated IgE levels underwent an oral challenge with portunus trituberculatus. Serum samples were collected from 33 children with a positive oral food challenge （test group） and from 30 health check-up child examinees （control group）. Total proteins were extracted from fresh portunus trituberculatus. Sodium dodecyl sulfate polyacrylamide gel electrophoresis （SDS-PAGE） and Western blot were conducted to identify the protein fragments of portunus trituberculatus responsible for AD among these children. Results The SDS-PAGE of crude protein extracts from portunus trituberculatus yielded 11 protein bands with relative molecular masses of 94 000, 70 000, 58 000, 49 000, 36 000, 34 000, 32 000, 27 000, 21 000, 19 000 and 17 000 respectively. Of the 11 protein bands, only 4 with relative molecular masses of 70 000, 58 000, 49 000 and 36 000 respectively reacted with sera from the patients by Western blot, with the reaction rate being 93.9%, 45.4%, 39.4% and 100% respectively. None of these protein bands reacted with sera from the control group by Western blot. There were significant differences between the test group and control group in the reaction rates of the four proteins with relative molecular masses of 70 000, 58 000, 49 000 and 36 000 respectively to sera （χ2 = 55.483, 17.898, 14.891, 63.000, all P < 0.05）. Conclusion The two proteins with relative molecular masses of 70 000 and 36 000 respectively are major allergens in portunus trituberculatus responsible for AD among children.