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针对主要猫过敏原Fel d I的单克隆抗体。I. 亲和纯化的Fel d I和去除Fel d I的提取物的血清学及生物学活性。

Monoclonal antibodies to the major feline allergen Fel d I. I. Serologic and biologic activity of affinity-purified Fel d I and of Fel d I-depleted extract.

作者信息

de Groot H, van Swieten P, van Leeuwen J, Lind P, Aalberse R C

机构信息

Central Laboratory, The Netherlands Red Cross Blood Transfusion Service, Amsterdam.

出版信息

J Allergy Clin Immunol. 1988 Nov;82(5 Pt 1):778-86. doi: 10.1016/0091-6749(88)90079-6.

Abstract

Monoclonal antibodies were raised against a major feline allergen, Fel d I. The specificity of the antibodies for Fel d I was demonstrated by a modified crossed immunoelectrophoretic procedure. These antibodies were used in affinity purification and depletion. Fed d I was eluted from the affinity matrix at a low pH. It was tested in serologic and biologic assays. Results of both the RAST and the histamine release test with affinity-purified Fel d I confirmed that it is a very potent allergen for the majority of patients allergic to cats. In the extract depleted with the monoclonal antibody, the ratio Fel d I/cat albumin was reduced by a factor of 20. With this depleted extract, 18 of 25 patients showed a reduction greater than 50% of the bound anti-IgE in the RAST compared with the RAST with crude cat extract. In the histamine release test with cells from two patients, the activity of the depleted extract was reduced by factors of 30 and 40, respectively. Depletion of Fel d I by polyclonal antibodies resulted in a reduction of the ratio Fel d I/cat albumin by a factor of 330. In the RAST, a reduction of the IgE response greater than 50% was found in 23 of 25 patients; in 12 patients the reduction was greater than 90%. In the histamine release test, the polyclonally depleted extract was 200 to 300 times less potent than the crude cat-dander extract. These results indicate that a very large proportion of the allergenic activity of cat-dander extract is caused by a single molecule: Fel d I.

摘要

制备了针对主要猫过敏原Fel d I的单克隆抗体。通过改良的交叉免疫电泳方法证明了这些抗体对Fel d I的特异性。这些抗体用于亲和纯化和去除。Fel d I在低pH值下从亲和基质上洗脱下来。对其进行了血清学和生物学检测。用亲和纯化的Fel d I进行的RAST和组胺释放试验结果均证实,对于大多数对猫过敏的患者来说,它是一种非常强效的过敏原。在用单克隆抗体去除的提取物中,Fel d I/猫白蛋白的比例降低了20倍。使用这种去除后的提取物,25名患者中有18名在RAST中显示与粗猫提取物相比,结合的抗IgE减少了50%以上。在对两名患者的细胞进行的组胺释放试验中,去除后的提取物活性分别降低了30倍和40倍。用多克隆抗体去除Fel d I导致Fel d I/猫白蛋白的比例降低了330倍。在RAST中,25名患者中有23名的IgE反应降低超过50%;12名患者的降低超过90%。在组胺释放试验中,多克隆去除后的提取物效力比粗猫皮屑提取物低200至300倍。这些结果表明,猫皮屑提取物中很大一部分过敏活性是由单一分子Fel d I引起的。

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