Monoclonal IgE antibodies against birch pollen allergens: Novel tools for biological characterization and standardization of allergens
Monoclonal IgE antibodies against birch pollen allergens: Novel tools for biological characterization and standardization of allergens
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DOI：
10.1067/mai.2003.1510
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发表时间：
2003-06-01
影响因子：
14.2
通讯作者：
Hoffmann, A
中科院分区：
文献类型：
--
作者：
Kaul, S;Scheurer, S;Hoffmann, A
背景：IgE抗体是速发型超敏反应的关键参与者。过敏原的特征和标准化通常是基于过敏患者的血清，而特异性的单克隆IgE抗体的临床相关allergens.Objective：本研究的目的是建立特异性的IgE单克隆抗体桦树花粉过敏原，因为这些是重要的吸入性过敏原。产生IgE的杂交瘤通过使用高灵敏度的大鼠嗜碱性白血病细胞介质释放试验来鉴定，通过额外的交叉刺激来增强过敏原刺激。与桦树花粉特异性IgG抗体连接。结果：获得了7株特异于桦树花粉变应原Betv 1或Betv 6的IgE单克隆抗体，在肥大细胞检测中均具有生物学活性。用mAb组合进行的介体释放实验表明，在Bet v 1上识别2个不同的表位区域，而2个Bet v 6特异性mAb结合至相同的表位区域。致敏后的大鼠嗜碱性白血病细胞与IgE单克隆抗体，不同量的Bet V 1或Bet V 6的检测在商业诊断过敏原试剂，而致敏与多克隆IgE导致类似的过敏原效力的所有products.Conclusions：IgE单克隆抗体代表有前途的新工具过敏原表征和组件解决标准化的过敏原提取物。
Background: IgE antibodies are key players in immediate hypersensitivity reactions. Allergen characterization and standardization is usually based on the sera of allergic patients, whereas monoclonal IgE antibodies specific for clinically relevant allergens are very rare.Objective: The aim of this study was to establish IgE mAbs specific for birch pollen allergens, because these are important inhalant allergens.Methods: IgE-producing hybridomas were identified by using the highly sensitive rat basophilic leukemia cell mediator release assay with enhanced allergen stimulation by additional cross-linking with birch pollen-specific IgG antibodies. The obtained IgE mAbs were characterized by immunologic methods and by cDNA sequencing.Results: Seven IgE mAbs specific for the birch pollen allergens Bet v 1 or Bet v 6 were obtained and were all biologically active in mast cell-based assays. Mediator release experiments with mAb combinations indicated that 2 different epitope regions were recognized on Bet v 1, whereas the 2 Bet v 6-specific mAbs bound to the same epitope region. After sensitization of rat basophilic leukemia cells with IgE mAbs, different amounts of Bet v 1 or Bet v 6 were detected in commercial diagnostic allergen reagents, whereas sensitization with polyclonal IgE resulted in similar allergenic potency of all products.Conclusions: IgE mAbs represent promising novel tools for allergen characterization and component-resolved standardization of allergen extracts.