Department of Applied Molecular Biosciences, Graduate School of Bioagricultural Sciences, Nagoya University, Chikusa-ku, Nagoya 464-8601, Japan.
Immunol Lett. 2010 Sep 6;133(1):28-34. doi: 10.1016/j.imlet.2010.06.007. Epub 2010 Jun 25.
Few common carbohydrate epitopes consisting of terminal beta-(1,2)-xylose and/or alpha-(1,3)-fucose residues are shared by a variety of glycoproteins from plants, insects and parasitic worms, termed cross-reactive carbohydrate determinant (CCD), and frequently recognized by IgE antibodies of patients with food and/or respiratory allergy, though clinical relevancy of such CCD-specific IgE is still controversial. Attention has also been focused on CCDs from the undesired post-translational modification of recombinant therapeutic proteins produced by transgenic plants and insects. In the present study, to clarify immunogenic potentials of CCD-bearing glycoproteins, the antibody response to a model plant glycoprotein, horseradish peroxidase (HRP) was investigated in a mouse model. C3H/He mice were immunized with HRP plus Al(OH)(3) or Freund's adjuvant, and IgG and IgE responses to CCDs in addition to HRP were analyzed by ELISA using some distinct glycoproteins with known N-glycan structures. IgE response to HRP was induced remarkably, whereas that to CCD was weaker and delayed. Moreover, apparent ratio of the CCD-specific antibodies to HRP-specific ones tended to be higher in IgG2a and IgG2b isotypes than IgG1, IgG3 and IgE. In contrast to rabbit antibodies, the CCD-specific antibodies from the mice gave poor reactivity with bromelain and honeybee phospholipase A2, suggesting the critical role of both beta-(1,2)-xylose and alpha-(1,3)-mannose in the CCD-recognition by the mouse antibodies. Moreover, the mouse antibodies showed weaker cross-reactivity to pollen- and insect-derived glycoproteins than the rabbit ones. Thus, in this mouse model, not only IgE but also IgG2 antibody responses to CCDs were induced by immunizing with a CCD-bearing glycoprotein, suggesting that CCDs affected not only Th2-type but also Th1-type antibody response at least in C3H/He mice.
几种常见的碳水化合物表位由末端β-(1,2)-木糖和/或α-(1,3)-岩藻糖残基组成,存在于植物、昆虫和寄生虫的各种糖蛋白中,被称为交叉反应性碳水化合物决定簇(CCD),经常被食物和/或呼吸过敏患者的 IgE 抗体识别,尽管这种 CCD 特异性 IgE 的临床相关性仍存在争议。人们也关注了由转基因植物和昆虫产生的重组治疗蛋白的非预期翻译后修饰所导致的 CCD。在本研究中,为了阐明含有 CCD 的糖蛋白的免疫原性潜力,我们在小鼠模型中研究了模型植物糖蛋白辣根过氧化物酶(HRP)的抗体反应。用 HRP 加 Al(OH)(3)或弗氏佐剂免疫 C3H/He 小鼠,并用一些具有已知 N-糖链结构的特异糖蛋白通过 ELISA 分析除 HRP 以外的对 CCD 的 IgG 和 IgE 反应。HRP 的 IgE 反应显著诱导,而 CCD 的较弱且延迟。此外,在 IgG2a 和 IgG2b 同种型中,CCD 特异性抗体与 HRP 特异性抗体的比值明显高于 IgG1、IgG3 和 IgE。与兔抗体不同,来自小鼠的 CCD 特异性抗体与菠萝蛋白酶和蜜蜂磷脂酶 A2 的反应性较差,表明在小鼠抗体识别 CCD 时,β-(1,2)-木糖和α-(1,3)-甘露糖均起关键作用。此外,与兔抗体相比,小鼠抗体对花粉和昆虫衍生糖蛋白的交叉反应性较弱。因此,在该小鼠模型中,用含有 CCD 的糖蛋白免疫不仅诱导了 IgE,还诱导了 IgG2 抗体对 CCD 的反应,这表明 CCD 不仅影响 Th2 型,而且至少在 C3H/He 小鼠中还影响 Th1 型抗体反应。