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俄罗斯刺沙蓬(猪毛菜)花粉提取物的免疫化学特性。变应原Sal k 1的纯化。

Immunochemical characterization of Russian thistle (Salsola kali) pollen extracts. Purification of the allergen Sal k 1.

作者信息

Carnés J, Fernández-Caldas E, Marina A, Alonso C, Lahoz C, Colás C, Lezaun A

机构信息

C.B.F. LETI, S.A. Research Laboratories, Tres Cantos, Madrid, Spain.

出版信息

Allergy. 2003 Nov;58(11):1152-6. doi: 10.1034/j.1398-9995.2003.00269.x.

Abstract

BACKGROUND

Salsola kali (Russian thistle) is a weed which belongs to the Chaenopodiacea family. It is widely distributed along the coasts of Europe, North Africa, USA and Australia. The objectives of this study were to study the allergenic composition of S. kali pollen and to purify an important allergen from the pollen extracts of this plant.

METHODS

A population of 66 individuals with specific IgE-mediated allergic symptoms and positive skin tests to S. kali were included in the study. Specific IgE to S. kali was determined by direct enzyme-linked immunosorbent assay (ELISA). The antigenic and allergenic profile of S. kali was evaluated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), isoelectric focussing (IEF) and immunoblot. Allergen purification was conducted by preparative SDS-PAGE. The allergenicity of the protein was evaluated by skin testing, direct ELISA, ELISA inhibition and immunoblots.

RESULTS

Specific IgE to S. kali was detected in 39 of the 66 individuals (59%). An allergen with a molecular weight of approximately 43 kDa was purified. This allergen was termed Sal k 1. A partial sequencing was obtained and no homology was found with other known proteins/allergens. The allergenicity of Sal k 1 was tested in vitro and in vivo. Of the 39 individuals with a positive specific IgE determination to S. kali, 26 (66.6%) had detectable specific IgE to Sal k 1. Twenty of these 39 individuals were skin-prick tested with the purified allergen (0.5 mg/ml) and all of them had a positive skin test to the purified allergen. Ten additional individuals, used as negative controls, had a negative response.

CONCLUSIONS

Sal k 1, an important allergen of S. kali, is recognized, in vitro, by approximately 67% of the patients sensitized to S. kali. Twenty patients with a positive skin test to a standardized S. kali extract had a positive reaction to the purified allergen.

摘要

背景

猪毛菜属于藜科杂草。它广泛分布于欧洲、北非、美国和澳大利亚的沿海地区。本研究的目的是研究猪毛菜花粉的致敏成分,并从该植物的花粉提取物中纯化一种重要的过敏原。

方法

本研究纳入了66名对猪毛菜有特异性IgE介导的过敏症状且皮肤试验呈阳性的个体。通过直接酶联免疫吸附测定(ELISA)测定对猪毛菜的特异性IgE。采用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)、等电聚焦(IEF)和免疫印迹法评估猪毛菜的抗原性和致敏原谱。通过制备性SDS-PAGE进行过敏原纯化。通过皮肤试验、直接ELISA、ELISA抑制和免疫印迹法评估该蛋白的致敏性。

结果

66名个体中有39名(59%)检测到对猪毛菜的特异性IgE。纯化出一种分子量约为43 kDa的过敏原。该过敏原被命名为Sal k 1。获得了部分序列,未发现与其他已知蛋白质/过敏原具有同源性。在体外和体内对Sal k 1的致敏性进行了测试。在39名对猪毛菜特异性IgE测定呈阳性的个体中,26名(66.6%)检测到对Sal k 1的特异性IgE。这39名个体中有20名用纯化的过敏原(0.5 mg/ml)进行了皮肤点刺试验,他们对纯化的过敏原皮肤试验均呈阳性。另外10名用作阴性对照的个体反应为阴性。

结论

Sal k 1是猪毛菜的一种重要过敏原,在体外,约67%对猪毛菜致敏的患者可识别该过敏原。20名对标准化猪毛菜提取物皮肤试验呈阳性的患者对纯化的过敏原反应呈阳性。

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