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萌发前后11种真菌的过敏原检测

Allergen detection from 11 fungal species before and after germination.

作者信息

Green Brett James, Mitakakis Teresa Zinovia, Tovey Euan Roger

机构信息

Faculty of Medicine, Woolcock Allergen Unit, Room 461, Blackburn Building D06, University of Sydney, NSW, Australia 2006.

出版信息

J Allergy Clin Immunol. 2003 Feb;111(2):285-9. doi: 10.1067/mai.2003.57.

Abstract

BACKGROUND

Allergens dispersed by airborne fungal spores play an important but poorly understood role in the underlying cause and exacerbation of asthma. Previous studies suggest that spores of Alternaria and Aspergillus release greater quantities of allergen after germination than before germination. It is unknown whether this is true of other allergenic fungi.

OBJECTIVE

Our purpose was to investigate the release of allergen from a range of individual fungal spores before and after germination.

METHODS

Allergen expression from spores of Alternaria alternata, Cladosporium herbarum, Aspergillus fumigatus, Botrytis cinerea, Epicoccum nigrum, Exserohilum rostratum, Penicillium chrysogenum, Stemphylium botryosum, Curvularia lunata, Trichoderma viride, and Bipolaris spicifera was examined by halogen immunoassays through the use of pooled serum IgE from patients allergic to fungus. Spores were deposited onto protein-binding membranes direct from culture. To germinate spores, samples were incubated in high humidity at room temperature for 48 hours. Ungerminated and germinated samples were then laminated with an adhesive film and immunostained by the halogen assay. The samples were examined by light microscopy, and positive counts (haloed particles) were expressed as percentages of total spores.

RESULTS

For 9 of 11 species, between 5.7% and 92% of spores released allergen before germination. Spores of Penicillium and Trichoderma did not release detectable allergen. After germination, all spores that germinated had allergen elution from their hyphae. Eight of 11 species showed a significant increase (P <.05) in the percentage of spores eluting detectable allergen. Localization of allergen along the hyphae varied with species, such that some eluted allergen mainly from hyphal tips and septal junctions whereas others eluted allergen along the entire length.

CONCLUSIONS

Increased elution of allergen after germination might be a common feature of many species of allergenic fungi. Although allergens from both spores and hyphae were recognized by human IgE, the extent to which human exposure occurs to allergens eluted from inhaled spores or from hyphae that germinate after deposition in the respiratory tract remains to be explored. The patterns of allergen expression might affect the clinical response to such exposure.

摘要

背景

空气中真菌孢子散播的过敏原在哮喘的潜在病因及病情加重方面起着重要作用,但人们对此了解甚少。先前的研究表明,链格孢属和曲霉属的孢子在萌发后释放的过敏原量比萌发前更多。其他致敏真菌是否也是如此尚不清楚。

目的

我们的目的是研究一系列单个真菌孢子在萌发前后的过敏原释放情况。

方法

通过使用对真菌过敏患者的混合血清IgE,采用卤素免疫测定法检测链格孢、草本枝孢、烟曲霉、灰葡萄孢、黑附球菌、喙突脐蠕孢、产黄青霉、匐柄霉、新月弯孢、绿色木霉和稻瘟平脐蠕孢孢子的过敏原表达。将孢子直接从培养物中沉积到蛋白质结合膜上。为使孢子萌发,将样品在室温高湿度条件下孵育48小时。然后将未萌发和已萌发的样品用胶膜覆盖,并通过卤素测定法进行免疫染色。通过光学显微镜检查样品,阳性计数(有晕圈的颗粒)以占总孢子的百分比表示。

结果

11个物种中的9个,5.7%至92%的孢子在萌发前释放过敏原。青霉属和木霉属的孢子未释放可检测到的过敏原。萌发后,所有萌发的孢子其菌丝都有过敏原洗脱。11个物种中的8个显示,洗脱可检测到过敏原的孢子百分比显著增加(P<.05)。过敏原沿菌丝的定位因物种而异,有些主要从菌丝尖端和隔膜连接处洗脱过敏原,而另一些则沿整个长度洗脱过敏原。

结论

萌发后过敏原洗脱增加可能是许多致敏真菌物种共有的特征。虽然孢子和菌丝的过敏原都能被人IgE识别,但人体吸入的孢子或呼吸道沉积后萌发的菌丝所洗脱的过敏原,人体接触的程度仍有待探索。过敏原表达模式可能会影响对此类接触的临床反应。

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