Wagner S, Bublin M, Hafner C, Kopp T, Allwardt D, Seifert U, Arif S A, Scheiner O, Breiteneder H
Department of Pathophysiology, Center for Physiology and Pathophysiology, Medical University of Vienna, Vienna, Austria.
Int Arch Allergy Immunol. 2007;143(4):246-54. doi: 10.1159/000100569. Epub 2007 Mar 9.
The latex of Hevea brasiliensis trees contains a complex proteome that includes a range of allergenic proteins. Current latex extracts that are used for the diagnosis of latex allergy still lack important allergens. We aimed to devise a production process for an improved reagent that would ideally contain the complete latex allergome.
Latex C-serum was fractionated by ammonium sulfate precipitation, and B- and C-serum proteins were then separated by anion exchange chromatography. Proteins eluting within defined salt concentration ranges were pooled into six final fractions. Fractions were evaluated by two-dimensional electrophoresis and subsequent IgE immunoblot for their spectrum of allergens. The presence of the most important latex allergens in the fractions was checked by Western blot analyses. Each fraction was further evaluated by skin prick test (SPT).
Reproducibility of the preparation method was demonstrated with two batches of latex. Comparison of latex B- and C-serum to the six fractions showed a remarkable increase in the number of detectable allergens in the fractions. The presence of the latex allergens Hev b 1-8 and Hev b 13 in the fractions was demonstrated. In SPTs, the fractions produced wheal-and-flare reactions comparable to commercial latex extracts.
This method provides reproducible latex protein fractions of high allergen content for the diagnosis of latex allergy.
巴西橡胶树的乳胶含有复杂的蛋白质组，其中包括一系列过敏原蛋白。目前用于乳胶过敏诊断的乳胶提取物仍缺乏重要的过敏原。我们旨在设计一种生产工艺，以制备一种理想情况下包含完整乳胶过敏原组的改进试剂。
通过硫酸铵沉淀对乳胶C血清进行分级分离，然后通过阴离子交换色谱法分离B血清和C血清蛋白。在规定盐浓度范围内洗脱的蛋白质被合并为六个最终级分。通过二维电泳和随后的IgE免疫印迹评估级分的过敏原谱。通过蛋白质印迹分析检查级分中最重要的乳胶过敏原的存在情况。每个级分进一步通过皮肤点刺试验（SPT）进行评估。
用两批乳胶证明了制备方法的可重复性。将乳胶B血清和C血清与六个级分进行比较，结果显示级分中可检测到的过敏原数量显著增加。证明了级分中存在乳胶过敏原Hev b 1-8和Hev b 13。在皮肤点刺试验中，这些级分产生的风团和红晕反应与市售乳胶提取物相当。
该方法为乳胶过敏的诊断提供了具有高过敏原含量的可重复乳胶蛋白级分。
