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嗜干真菌青霉状曲霉粗菌丝体提取物免疫球蛋白E结合蛋白的鉴定及不同过敏原反应性谱受试者的血清学反应性评估

Identification of Immunoglobulin E-Binding Proteins of the Xerophilic Fungus Aspergillus penicillioides Crude Mycelial Mat Extract and Serological Reactivity Assessment in Subjects with Different Allergen Reactivity Profiles.

作者信息

González De León Joenice, González Méndez Ricardo, Cadilla Carmen L, Rivera-Mariani Félix E, Bolaños-Rosero Benjamín

机构信息

Department of Microbiology and Medical Zoology, School of Medicine, University of Puerto Rico, San Juan, PR, USA.

出版信息

Int Arch Allergy Immunol. 2018;175(3):147-159. doi: 10.1159/000484898. Epub 2018 Feb 3.

Abstract

BACKGROUND

Aspergillus penicillioides is a very common indoor xerophilic fungus and potential causative agent of respiratory conditions. Although people are constantly exposed to A. penicillioides, no proteins with allergenic potential have been described. Therefore, we aim to confirm allergic sensitization to A. penicillioides through reactivity in serological assays and detect immunoglobulin E (IgE)-binding proteins.

METHODS

In an indirect ELISA, we compared the serological reactivity to A. penicillioides between subjects with specific IgE (sIgE) (group 1, n = 54) and no sIgE reactivity (group 2, n = 15) against commercial allergens. Correlations and principal component analysis were performed to identify associations between reactivity to commercial allergens and A. penicillioides. IgE-binding proteins in A. penicillioides were visualized using Western blotting (WB) in group 1. The IgE-binding proteins with the highest reactivity were analyzed by mass spectrometry and confirmed by transcript matching.

RESULTS

There was no statistical significance (p = 0.1656) between the study groups in serological reactivity. Correlations between reactivity to A. penicillioides, dog epithelia, Aspergillus fumigatus, and Penicillium chrysogenum were observed. WB experiments showed 6 IgE-binding proteins with molecular weights ranging from 45 to 145 kDa. Proteins of 108, 83, and 56 kDa showed higher reactivity. Mass spectrometry analysis of these 3 proteins led to the putative identification of NADP-specific glutamate dehydrogenase and catalase B. This was confirmed with transcriptome analysis.

CONCLUSIONS

These results provide evidence of the presence of potential allergenic components in A. penicillioides. Further analysis of the putatively identified proteins should reveal their allergenic potential.

摘要

背景

青霉状曲霉是一种非常常见的室内嗜干性真菌,也是呼吸道疾病的潜在病原体。尽管人们经常接触青霉状曲霉,但尚未有具有致敏潜力的蛋白质被描述。因此,我们旨在通过血清学检测中的反应性来确认对青霉状曲霉的过敏致敏情况,并检测免疫球蛋白E(IgE)结合蛋白。

方法

在间接酶联免疫吸附测定(ELISA)中,我们比较了对商业过敏原具有特异性IgE(sIgE)的受试者(第1组,n = 54)和无sIgE反应性的受试者(第2组,n = 15)之间对青霉状曲霉的血清学反应性。进行相关性分析和主成分分析,以确定对商业过敏原和青霉状曲霉的反应性之间的关联。在第1组中,使用蛋白质印迹法(WB)观察青霉状曲霉中的IgE结合蛋白。对反应性最高的IgE结合蛋白进行质谱分析,并通过转录本匹配进行确认。

结果

研究组之间的血清学反应性无统计学意义(p = 0.1656)。观察到对青霉状曲霉、狗上皮、烟曲霉和产黄青霉的反应性之间存在相关性。WB实验显示6种IgE结合蛋白,分子量范围为45至145 kDa。分子量为108、83和56 kDa的蛋白质显示出更高的反应性。对这3种蛋白质的质谱分析导致推测鉴定出NADP特异性谷氨酸脱氢酶和过氧化氢酶B。转录组分析证实了这一点。

结论

这些结果提供了青霉状曲霉中存在潜在过敏原成分的证据。对推测鉴定出的蛋白质的进一步分析应揭示其致敏潜力。

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Fungi and allergic lower respiratory tract diseases.真菌与过敏性下呼吸道疾病。
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