Schwienbacher M, Israel L, Heesemann J, Ebel F
Max-von-Pettenkofer-Institute, Ludwig-Maximilans-University, Munich, Germany.
Allergy. 2005 Nov;60(11):1430-5. doi: 10.1111/j.1398-9995.2005.00904.x.
Aspergillus fumigatus is a pathogenic mould causing allergic and invasive respiratory diseases. Allergic bronchopulmonary Aspergillosis (ABPA) is a severe pulmonary complication resulting from hypersensitivity to A. fumigatus proteins. Aspergillus allergen Asp f6 is recognized by IgE from ABPA patients, but not from sensitized individuals, a fact that can be used to differentiate between these two groups of allergic patients.
Proteins from hyphae, resting and germinating conidia of A. fumigatus were compared by SDS-PAGE. Protein identification was performed using MALDI-TOF mass spectrometry. Recombinant A. fumigatus allergens were used to isolate specific monoclonal antibodies (mab) from a hybridoma bank generated against Aspergillus proteins.
A hyphae-specific 23 kDa A. fumigatus protein was identified as the allergen Asp f6/manganese-dependent superoxide dismutase (MnSOD). Differential expression of MnSOD was confirmed by immunoblot using a specific mab. In contrast, Asp f8 another intracellular, but not ABPA-specific allergen, was detected in hyphae and conidia.
Aspergillus fumigatus is able to colonize its environment by the formation of hyphae. Hyphae are found in the lung of ABPA patients, but not in patients suffering from atopic asthma. Our finding that Asp f6 is specifically expressed in hyphae might explain why an IgE response to Asp f6 is specific for ABPA patients.
烟曲霉是一种致病性霉菌,可引起过敏性和侵袭性呼吸道疾病。变应性支气管肺曲霉病(ABPA)是对烟曲霉蛋白过敏导致的严重肺部并发症。烟曲霉变应原Asp f6可被ABPA患者的IgE识别,但不能被致敏个体的IgE识别,这一事实可用于区分这两组过敏性患者。
通过SDS-PAGE比较烟曲霉的菌丝体、静止和萌发分生孢子中的蛋白质。使用MALDI-TOF质谱进行蛋白质鉴定。使用重组烟曲霉变应原从针对曲霉蛋白产生的杂交瘤库中分离特异性单克隆抗体(mab)。
一种菌丝体特异性的23 kDa烟曲霉蛋白被鉴定为变应原Asp f6/锰依赖性超氧化物歧化酶(MnSOD)。使用特异性mab通过免疫印迹证实了MnSOD的差异表达。相比之下,另一种细胞内但非ABPA特异性变应原Asp f8在菌丝体和分生孢子中被检测到。
烟曲霉能够通过形成菌丝体在其环境中定殖。菌丝体存在于ABPA患者的肺部,但不存在于特应性哮喘患者的肺部。我们发现Asp f6在菌丝体中特异性表达,这可能解释了为什么对Asp f6的IgE反应对ABPA患者具有特异性。