中华皮肤科杂志 ›› 2011, Vol. 44 ›› Issue (2): 84-87.

• 论著 • 上一篇    下一篇

人源性抗大疱性类天疱疮抗原BP180单链抗体的体外功能鉴定

吴岩1,孙娜娜2,史晓蔚2,呼蕾2,李春英3,王刚1   

  1. 1. 第四军医大学西京医院全军皮肤病研究所
    2.
    3. 西安第四军医大学西京医院皮肤科
  • 收稿日期:2010-08-06 修回日期:2010-09-08 出版日期:2011-02-15 发布日期:2011-02-10
  • 通讯作者: 王刚 E-mail:xjwgang@fmmu.edu.cn
  • 基金资助:

    国家自然科学基金;教育部留学回国人员启动基金

In vitro functional characterization of human anti-bullous pemphigoid 180 （BP180） single-chain Fv antibody

  • Received:2010-08-06 Revised:2010-09-08 Online:2011-02-15 Published:2011-02-10
  • Contact: WANG Gang E-mail:xjwgang@fmmu.edu.cn

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摘要：

目的 探讨人源性抗大疱性类天疱疮抗原BP180单链抗体的生物学功能。方法 亲和层析方法纯化大疱性类天疱疮（BP）患者血清自身抗体，通过竞争性ELISA、竞争性免疫荧光和补体活化的竞争性抑制实验来观察所制备的抗BP180单链抗体对BP-IgG自身抗体结合人BP180抗原的竞争性抑制作用。结果 竞争性ELISA结果表明，在0 ~ 60 μg/ml范围内单链抗体对BP-IgG自身抗体的竞争性抑制作用呈剂量依赖关系，最大抑制率可达69.50%（与对照组相比，均有统计学意义，P < 0.01）。竞争性免疫荧光实验中，单链抗体浓度增加至40 μg/ml时，BP-IgG在基底膜带的沉积及其介导的补体C3活化作用均被抑制，呈阴性。结论 人源性抗BP180单链抗体在体外对BP致病性自身抗体与抗原结合及后续的补体活化具有一定的抑制作用。

关键词: 抑制

Abstract:

Objective To characterize the function of human anti-BP180 single-chain Fv antibody （scFv） in vitro. Methods The IgG autoantibodies against BP180 were purified by affinity chromatography from the sera of patients with BP. The inhibitive effect of previously constructed anti-BP180 scFv on the binding of anti-BP180 IgG autoantibodies to the recombinant NC16A domain of human BP180 antigen was observed by competitive ELISA, competitive immunofluorescence assay and competitive inhibition test for complement activation. Results As ELISA revealed, the scFv significantly inhibited the binding of anti-BP180 IgG autoantibodies to the corresponding antigen （P < 0.01）, and the inhibitive effect was dose-dependent within the concentration range from 0 to 60 μg/ml. The inhibitive rate peaked at 69.50%. The deposition of anti-BP180-IgG autoantibodies in basement membrane zone and the IgG autoantibody-mediated complement C3 activation were both suppressed by the scFv of 40 μg/ml. Conclusion The genetically engineered anti-BP180 scFv has a certain inhibitive effect on the binding of BP-IgG autoantibodies to BP180 antigen and on the subsequent complement activation in vitro.

Key words: Inhibition