Horner W E, Ibanez M D, Liengswangwong V, Salvaggio J E, Lehrer S B
Tulane University School of Medicine, Department of Medicine, New Orleans, La. 70112.
J Allergy Clin Immunol. 1988 Dec;82(6):978-86. doi: 10.1016/0091-6749(88)90134-0.
Crude extracts of Pleurotus ostreatus spores obtained from a single local source were fractionated by gel filtration to resolve the allergenic components. The fraction pool corresponding to 10.5 to 25 kd molecular weight contained allergenic activity as demonstrated by both RAST and skin testing. Similar results were obtained with extracts from spores that originated in four other areas and with extracts prepared from P. sajor-caju spores obtained from commercially produced caps. The RAST-active fraction was further separated by hydrophobic interaction chromatography (HIC). HIC fraction pools were assayed for allergen(s) by RAST inhibition and immunoblotting of isoelectric focused polyacrylamide gels. RAST-inhibition data indicated that the allergen(s) was reversibly bound to the HIC column, eluting with 2, 1, and 0.15 mol/L of buffered salt solutions. After electrofocusing, these fractions yielded 15, 12, and 11 Coomassie brilliant blue-staining bands, respectively. IgE binding occurred with 7, 8, and 6 of these bands, as revealed by radiostaining of the immunoblots. These procedures help identify P. ostreatus spore allergens and allow a greater degree of standardization in the preparation of allergen extracts from basidiospores for use in diagnosis and therapy of fungal allergy.
从单一本地来源获取的平菇孢子粗提物通过凝胶过滤进行分级分离，以解析其致敏成分。通过放射性过敏原吸附试验（RAST）和皮肤试验证明，分子量在10.5至25kd之间的分级组分池具有致敏活性。来自其他四个地区的孢子提取物以及从商业生产的菌盖中获得的凤尾菇孢子制备的提取物也得到了类似结果。通过疏水相互作用色谱法（HIC）对具有RAST活性的组分进行进一步分离。通过RAST抑制试验以及等电聚焦聚丙烯酰胺凝胶的免疫印迹法对HIC分级组分池进行过敏原检测。RAST抑制试验数据表明，过敏原与HIC柱可逆结合，用2、1和0.15mol/L的缓冲盐溶液洗脱。电聚焦后，这些组分分别产生15、12和11条考马斯亮蓝染色带。免疫印迹的放射染色显示，IgE与其中7、8和6条带发生结合。这些方法有助于鉴定平菇孢子过敏原，并在制备用于真菌过敏诊断和治疗的担子孢子过敏原提取物时实现更高程度的标准化。
