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不同弯孢属物种间交叉反应蛋白的鉴定。

Identification of cross-reactive proteins amongst different Curvularia species.

作者信息

Gupta Ratna, Singh Bhanu P, Sridhara Susheela, Gaur Shailendra N, Kumar Raj, Chaudhary Vijay K, Arora Naveen

机构信息

Centre for Biochemical Technology, University of Delhi South Campus, Delhi, India.

出版信息

Int Arch Allergy Immunol. 2002 Jan;127(1):38-46. doi: 10.1159/000048167.

Abstract

BACKGROUND

Curvularia lunata is an important inhalant allergen. The present study was undertaken to investigate the shared IgG- and IgE-binding components among seven Curvularia species prevalent in the aerospora.

METHODS

Seven different Curvularia species were grown in a semisynthetic medium for 13 days. The extracts were analyzed by SDS-PAGE, immunoblot and ELISA/immunoblot inhibition using sera from C. lunata-positive patients and anti-C. lunata rabbit serum.

RESULTS

Different Curvularia species showed 11-19 protein bands on SDS-PAGE. Proteins of 12, 20, 31, 45, 53, 78 and 97 kD were present in all the species. Eight out of 98 nasobronchial patients exhibited positive skin tests to C. lunata and to at least five Curvularia species. ELISA using these sera showed IgE binding with Curvularia species. Immunoblot using pooled anti-C. lunata sera from patients showed 5-12 allergenic proteins. Proteins of 12, 31, 45, 53 and 78 kD showed IgE binding in Curvularia species. Antibodies against C. lunata detected 6-14 antigenic proteins on immunoblot. Proteins of 31, 45 and 53 kD showed IgG binding in all the species. Proteins of 31 and 53 kD showed complete IgE/IgG binding inhibition. IgE/IgG ELISA inhibition showed dose-dependent inhibition in Curvularia species. C. lunata extract required 0.17 and 0.11 microg of protein for 50% IgE and IgG inhibition, respectively. C. clavata and C. pallescens required 10 times more protein to exhibit the same inhibition and other species required similar protein levels as those required by C. lunata.

CONCLUSIONS

A high degree of cross-reactivity was observed between C. lunata and the six other Curvularia species tested. C. lunata and C. senegalensis shared maximum allergenic and antigenic components.

摘要

背景

新月弯孢菌是一种重要的吸入性变应原。本研究旨在调查气传孢子中常见的七种弯孢菌属物种之间共享的IgG和IgE结合成分。

方法

七种不同的弯孢菌属物种在半合成培养基中培养13天。使用来自新月弯孢菌阳性患者的血清和抗新月弯孢菌兔血清,通过SDS-PAGE、免疫印迹和ELISA/免疫印迹抑制分析提取物。

结果

不同的弯孢菌属物种在SDS-PAGE上显示出11-19条蛋白带。所有物种中均存在12、20、31、45、53、78和97kD的蛋白质。98名鼻支气管患者中有8名对新月弯孢菌和至少五种弯孢菌属物种的皮肤试验呈阳性。使用这些血清的ELISA显示IgE与弯孢菌属物种结合。使用患者混合抗新月弯孢菌血清的免疫印迹显示5-12种变应原蛋白。12、31、45、53和78kD的蛋白质在弯孢菌属物种中显示IgE结合。抗新月弯孢菌抗体在免疫印迹上检测到6-14种抗原蛋白。31、45和53kD的蛋白质在所有物种中显示IgG结合。31和53kD的蛋白质显示出完全的IgE/IgG结合抑制。IgE/IgG ELISA抑制在弯孢菌属物种中显示出剂量依赖性抑制。新月弯孢菌提取物分别需要0.17和0.11μg蛋白质才能达到50%的IgE和IgG抑制。棒状弯孢菌和苍白弯孢菌需要多10倍的蛋白质才能表现出相同的抑制作用,其他物种需要与新月弯孢菌所需的蛋白质水平相似。

结论

在新月弯孢菌与其他六种测试的弯孢菌属物种之间观察到高度的交叉反应性。新月弯孢菌和塞内加尔弯孢菌共享最多的变应原和抗原成分。

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