Verma J, Gangal S V
Centre for Biochemical Technology, Delhi, India.
Int Arch Allergy Immunol. 1994;104(2):175-83. doi: 10.1159/000236727.
Allergenic components of the fungus Fusarium solani were isolated using (NH4)2SO4 precipitation and ion-exchange column chromatography. The allergenicity of fractions was studied by enzyme-linked immunosorbent assay and radioallergosorbent test inhibition techniques. Proteins of culture filtrate (CF), mycelium (MY), and spore (SP) extracts of F. solani were characterized by isoelectrofocusing, sodium dodecyl sulphate-polyacrylamide gel electrophoresis, and IgE-specific immunoblotting. CF antigen of F. solani contained more allergenic proteins than MY and SP, visible on immunoblot analysis using allergenic serum pool. A 65-kD protein component of CF was found to be a major allergen, as it was strongly visible on immunoblots of all 15 patient sera tested. Crossed immunoelectrophoresis and enzyme-linked immunosorbent assay inhibition using rabbit antibodies raised against F. solani CF demonstrated shared antigenicity between CF, MY, and SP extracts. It was observed that F. solani is a significant allergen, and most of the allergens of MY and SP extracts were found in CF extract. Therefore, CF alone can be used in the preparation of a standard extract. However, few unique allergenic proteins were observed in MY as well as in SP extracts of F. solani. Hence, the use of combined CF, MY, and SP extracts of F. solani is recommended for diagnosis and immunotherapy.
采用硫酸铵沉淀法和离子交换柱色谱法分离茄病镰刀菌的变应原成分。通过酶联免疫吸附测定和放射变应原吸附试验抑制技术研究各组分的变应原性。利用等电聚焦、十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和IgE特异性免疫印迹法对茄病镰刀菌培养滤液(CF)、菌丝体(MY)和孢子(SP)提取物中的蛋白质进行表征。茄病镰刀菌的CF抗原比MY和SP含有更多的变应原性蛋白质,在使用变应原血清库进行免疫印迹分析时可见。CF的一种65-kD蛋白质成分被发现是主要变应原,因为在所有15份受试患者血清的免疫印迹上都能强烈看到。使用针对茄病镰刀菌CF产生的兔抗体进行交叉免疫电泳和酶联免疫吸附测定抑制,结果表明CF、MY和SP提取物之间存在共同抗原性。据观察,茄病镰刀菌是一种重要的变应原,MY和SP提取物中的大多数变应原都存在于CF提取物中。因此,单独使用CF即可用于制备标准提取物。然而,在茄病镰刀菌的MY和SP提取物中也观察到了一些独特的变应原性蛋白质。因此,建议使用茄病镰刀菌的CF、MY和SP提取物组合进行诊断和免疫治疗。