Untersmayr Eva, Szalai Krisztina, Riemer Angelika B, Hemmer Wolfgang, Swoboda Ines, Hantusch Brigitte, Schöll Isabella, Spitzauer Susanne, Scheiner Otto, Jarisch Reinhart, Boltz-Nitulescu George, Jensen-Jarolim Erika
Center of Physiology and Pathophysiology, Medical University of Vienna, Waehringer Guertel 18-20, A-1090 Vienna, Austria.
Mol Immunol. 2006 Mar;43(9):1454-61. doi: 10.1016/j.molimm.2005.07.038. Epub 2005 Sep 15.
Parvalbumin, the major fish allergen, is recognized by allergen-specific IgE of more than 90% of all fish-allergic patients. A detailed knowledge of allergenic structures is crucial for developing a vaccine inducing blocking antibodies specifically directed towards the IgE binding epitopes. In the present study we aimed to use the phage display technique to generate mimotopes, which mimic epitopes on parvalbumin. Parvalbumin-specific IgE was purified from sera of fish-allergic patients and used for screening of a constrained decamer phage library. After four rounds of biopanning using parvalbumin-specific IgE, five phage clones were selected which were specifically recognized by parvalbumin-specific IgE as well as IgG. DNA sequencing and peptide alignment revealed a high degree of sequence similarities between the mimotopes. Interestingly, on the surface of natural parvalbumin three regions could be defined by computational mimotope matching. In accordance, previously defined allergenic peptides of cod parvalbumin highlighted areas in close proximity or overlapping with the mimotope matching sites. From the presented data we conclude that our approach identified conformational epitopes of parvalbumin relevant for IgE and IgG binding. We suggest that these mimotopes are suitable candidates for an epitope-specific immunotherapy of fish-allergic patients.
小白蛋白是主要的鱼类过敏原，超过90%的鱼类过敏患者的过敏原特异性IgE可识别该蛋白。详细了解过敏原结构对于开发一种能诱导产生特异性针对IgE结合表位的阻断抗体的疫苗至关重要。在本研究中，我们旨在利用噬菌体展示技术生成模拟小白蛋白表位的模拟表位。从小鱼类过敏患者血清中纯化出小白蛋白特异性IgE，并用于筛选受限的十肽噬菌体文库。使用小白蛋白特异性IgE进行四轮生物淘选后，选择了五个噬菌体克隆，它们能被小白蛋白特异性IgE以及IgG特异性识别。DNA测序和肽序列比对显示模拟表位之间存在高度的序列相似性。有趣的是，通过计算模拟表位匹配可在天然小白蛋白表面确定三个区域。相应地，先前确定的鳕鱼小白蛋白过敏原肽突出了与模拟表位匹配位点紧密相邻或重叠的区域。根据所呈现的数据，我们得出结论，我们的方法鉴定出了与IgE和IgG结合相关的小白蛋白构象表位。我们认为这些模拟表位是鱼类过敏患者表位特异性免疫疗法的合适候选物。
