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通过免疫印迹法鉴定古巴裸盖菇孢子过敏原

Identification of Psilocybe cubensis spore allergens by immunoprinting.

作者信息

Helbling A, Horner W E, Lehrer S B

机构信息

Department of Medicine, Tulane University School of Medicine, New Orleans, LA 70112.

出版信息

Int Arch Allergy Immunol. 1993;100(3):263-7. doi: 10.1159/000236422.

Abstract

Previous studies established that Psilocybe cubensis contains potent allergens, and that a significant percentage of atopic subjects were sensitized to P. cubensis spores. The objective of this study was to identify P. cubensis spore allergens using isoelectric focusing (IEF) and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) immunoprinting. Coomassie blue staining of IEF gels detected approximately 20 bands between pI 3.6 and 9.3. Immunoprints obtained with 15 P. cubensis skin test- and RAST-positive sera revealed 13 IgE-binding bands; the most reactive were at pI 5.0 (80%), 5.6 (87%), 8.7 (80%) and 9.3 (100%). SDS-PAGE resolved 27 proteins ranging from about 13 to 112 kD. SDS-PAGE immunoprints conducted with 11 skin test- and RAST-positive sera demonstrated 18 IgE-binding bands; most sera reacted to 16 (82%), 35 (100%) and 76 kD (91%) allergens. Both electrophoretic procedures demonstrated a single allergen (at pI 9.3 and 35 kD) that reacted with all sera tested. This study corroborates the allergenic significance of P. cubensis spores and identifies the allergens of greatest importance.

摘要

先前的研究证实,古巴裸盖菇含有强效过敏原,且相当比例的特应性个体对古巴裸盖菇孢子敏感。本研究的目的是使用等电聚焦(IEF)和十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)免疫印迹法鉴定古巴裸盖菇孢子过敏原。IEF凝胶的考马斯亮蓝染色在pI 3.6至9.3之间检测到约20条带。用15份古巴裸盖菇皮肤试验和放射性变应原吸附试验(RAST)阳性血清获得的免疫印迹显示13条IgE结合带;反应最强的位于pI 5.0(80%)、5.6(87%)、8.7(80%)和9.3(100%)。SDS-PAGE分离出27种蛋白质,分子量约为13至112 kD。用11份皮肤试验和RAST阳性血清进行的SDS-PAGE免疫印迹显示18条IgE结合带;大多数血清与16 kD(82%)、35 kD(100%)和76 kD(91%)的过敏原发生反应。两种电泳方法均显示一种单一过敏原(pI 9.3和35 kD)与所有测试血清发生反应。本研究证实了古巴裸盖菇孢子的致敏意义,并确定了最重要的过敏原。

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