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Molecular and immunological characterization of allergens from the entomopathogenic fungus Beauveria bassiana

机译:昆虫致病性球孢白僵菌的致敏原的分子和免疫学表征

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Background Entomopathogenic fungi such as Beauveria bassiana are considered promising biological control agents for a variety of arthropod pests. Beauveria species, however, have the potential to elicit allergenic reactions in humans, although no specific allergens have been characterized to date. Methods Four putative allergens were identified within B. bassiana expressed sequence tag (EST) datasets. IgE-reactivity studies were performed using sera from patients displaying mold allergies against recombinant B. bassiana proteins expressed in E. coli. Results Full length cDNA and genomic nucleotide sequences of four potential B. bassiana allergens were isolated. BLASTX search results led to their putative designation as follows; Bb-Eno1, with similarity to fungal enolases; Bb-f2, similar to the Aspergillus fumigatus major allergen, Asp f2 and to a fibrinogen binding mannoprotein; Bb-Ald, similar to aldehyde dehydrogenases; and Bb-Hex, similar to N-acetyl-hexosaminadases. All four genes were cloned into E. coli expression systems and recombinant proteins were produced. Immunoblots of E. coli extracts probed with pooled as well as individual human sera from patients displaying mould allergies demonstrated IgE reactivity versus recombinant Bb-Eno1 and Bb-Ald. Conclusion Four putative Beauveria bassiana allergens were identified. Recombinant proteins corresponding to two of the four, Bb-Eno1 and Bb-Ald were bound by sera IgEs derived from patients with fungal allergies. These data confirm the potential allergenicity of B. bassiana by identification of specific human IgE reactive epitopes.
机译:背景技术病原性真菌,例如球孢白僵菌,被认为是用于各种节肢动物害虫的有前途的生物防治剂。尽管目前尚未鉴定出特定的过敏原,但球孢白僵菌物种仍可能引起人类过敏反应。方法在球孢杆菌表达序列标签(EST)数据集中鉴定出四种假定的过敏原。 IgE反应性研究使用的患者血清显示出对大肠杆菌中表达的重组球孢杆菌蛋白质的霉菌过敏反应。结果分离出四种潜在的球孢杆菌过敏原的全长cDNA和基因组核苷酸序列。 BLASTX搜索结果导致其可能的名称如下: Bb-Eno1,与真菌烯醇类似; Bb-f2,类似于烟曲霉主要过敏原,Asp f2和与纤维蛋白原结合的甘露糖蛋白; Bb-Ald,类似于醛脱氢酶;和Bb-Hex,类似于N-乙酰-己糖胺酶。将所有四个基因克隆到大肠杆菌表达系统中,并产生重组蛋白。大肠杆菌霉菌提取物的免疫印迹用合并显示的霉菌过敏症患者的合并血清以及单个人血清进行检测,与重组Bb-Eno1和Bb-Ald相比,具有IgE反应性。结论鉴定了四种推定的球孢白僵菌过敏原。对应于四个Bb-Eno1和Bb-Ald中两个中的两个的重组蛋白被源自患有真菌过敏患者的血清IgE结合。这些数据通过鉴定特定的人IgE反应性表位证实了球孢杆菌的潜在变应原性。

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