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牛奶中高分子量蛋白致敏作用的初步研究

侯丽英   

  1. 天津医科大学临床免疫检验教研室
  • 收稿日期:2014-03-20 修回日期:2014-07-01 出版日期:2014-11-15 发布日期:2014-11-15
  • 通讯作者: 侯丽英

Preliminary Research on the Milk Allergy Induced by High Molecular Weight Protein

HOU Li ying   

  1. College of Medical Laboratory, Tianjin Medical University
  • Received:2014-03-20 Revised:2014-07-01 Published:2014-11-15 Online:2014-11-15
  • Contact: HOU Li ying

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摘要：

【摘要】 目的 探讨牛奶中高分子质量蛋白在牛奶过敏过程中的致敏作用。 方法 选取牛奶过敏患者血清 30例， 皮肤点刺试验阳性， 血清特异性 IgE（sIgE）检测结果均为阳性且≥1+。 健康对照血清 1 例， 血清 sIgE 检测阴性，无过敏史。 空腹采血， 收集血清于-20 ℃冻存备用。 利用 Sephadex G200 凝胶层析获得牛奶高分子质量蛋白， 通过蛋白免疫印迹法（Wetern blot）和酶联免疫吸附测定（ELISA）检测高分子质量蛋白与牛奶过敏患者血清 sIgE 的结合活性。 结果 SDS-PAGE 显示 Sephadex G200 凝胶层析纯化得到的高分子质量蛋白主要包括 3 条带， 分子质量约为67、80 和 160 ku， 推断其可能是牛血清白蛋白、乳铁蛋白和免疫球蛋白。 经 Western blot 分析， 3 种高分子质量蛋白与牛奶过敏患者血清均能反应， 并且在 67 ku 附近条带最明显。 经 ELISA 分析， 高分子质量蛋白与牛奶过敏患者血清的阳性反应率比β-乳球蛋白略高（分别是 46.7%、43.3%）。 结论 牛奶中高分子质量蛋白组分能够诱导特异性抗体产生， 这些组分在牛奶过敏过程中具有重要的致敏作用。

关键词: 食物过敏, 免疫球蛋白 E, 免疫印迹法, 酶联免疫吸附测定, 牛奶, 高分子质量蛋白, 致敏作用, 特异性 IgE

Abstract:  [Abstract]   Objective   To investigate the allergization of milk high molecular weight proteins.  Methods   Thirty cas?es of patients with serum allergic to milk were selected. Their skin prick tests were positive. Results of serum specific IgE (sIgE) test were positive and ≥ 1 +. One case of healthy control with negative sIgE test and without history of allertgy, was included in this study. The serum samples were collected and frozen at -20 ℃. Sephadex G200 gel chromatography was used to obtain milk high molecular weight proteins. Western blot and ELISA methods were used to detect milk high molecular weight proteins and the activity of serum sIgE.   Results   Results of SDS-PAGE showed that high molecular weights proteins displayed by Sephadex G200 gel chromatography, mainly including three bands, the molecular weight of 67 ku, 80 ku and 160 ku. Western blot analysis showed that three kinds of high molecular weights proteins can react with milk allergy serum, and the most obvious appeared near the molecular weight 67 ku band. ELISA analysis showed that the positive response rate of high molecular weight proteins with milk allergic patient’ s serum was slightly higher than that of β-lactoglobulin (46.7% and 43.3%, respectively).   Conclusion  The milk high molecular weight protein components can induce specific IgE antibodies, which have important sensitization in the process of milk allergy.

Key words: food hypersensitivity, immunoglobulin E, Western blotting, enzyme-linked immunosorbent assay, milk, high molecular weight protein, allergization, 特异性 IgE