两种与血管炎相关的角膜上皮衍生抗原的特性分析。

Characterization of two corneal epithelium-derived antigens associated with vasculitis.

作者信息

Reynolds I, John S L, Tullo A B, Ayad S, Morgan K, Ballardie F W, Holt P J, Hillarby M C

机构信息

Department of Rheumatology, University of Manchester, UK.

出版信息

Invest Ophthalmol Vis Sci. 1998 Dec;39(13):2594-601.

PMID:9856769

Abstract

PURPOSE

In a previous investigation into corneal autoimmunity, it was demonstrated that a putative autoantigen, a protein of 66 kDa, present in bovine corneal epithelium, binds circulating autoantibodies in approximately 60% of patients with Wegener's granulomatosis (WG). The aim of the present study was to characterize and identify the 66-kDa protein.

METHODS

A purification protocol was established for the 66-kDa protein using standard chromatography techniques. During the purification procedure it became clear that the 66-kDa protein detected in patients' sera was in fact two proteins, both running at 66 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, that eluted in different fractions on DE-52 chromatography columns. These two proteins have been labeled bovine corneal epithelial antigen-A and -B (BCEA-A and BCEA-B). Further investigations of antibody binding have demonstrated that patients' sera bind to either one or the other of these proteins with no cross-reactivity between them. Separated BCEA-A and BCEA-B protein extracts were immunoblotted with 27 WG patients' sera, 10 Churg-Strauss syndrome (CSS) patients' sera, 31 rheumatoid arthritis (RA) patients' sera, and 40 healthy control subjects' sera from the blood bank.

RESULTS

Forty-six percent of WG patients' sera had antibodies to one of the 66-kDa antigens, whereas none of the healthy control subjects' sera had 66-kDa antibodies (P < 10(-5)). In the WG group, 31% were positive to BCEA-A (versus controls, P = 0.0023), and 15% were positive to BCEA-B. WG patients with peripheral ulcerative keratitis (PUK) had a significant association with anti-BCEA-A antibodies when compared with healthy control subjects (50%, P < 10(-6)). However, in the RA group with no eye disease there was an association with BCEA-A (25%, P = 0.011) but not in the RA group with PUK. The frequency of anti-BCEA-B antibodies was significantly increased in patients with CSS (60%, P < 10(-7)).

CONCLUSIONS

In summary, it has been shown that vasculitis patients have antibodies to two 66-kDa corneal antigens and that autoantibodies to these antigens are mutually exclusive. It has also been shown that antibodies to BCEA-B are associated with CSS, whereas BCEA-A antibodies are associated with WG and RA.

摘要

目的

在先前一项关于角膜自身免疫的研究中，已证实牛角膜上皮中存在一种假定的自身抗原，即一种66 kDa的蛋白质，在约60%的韦格纳肉芽肿（WG）患者中可结合循环自身抗体。本研究的目的是对这种66 kDa蛋白质进行表征和鉴定。

方法

使用标准色谱技术建立了针对66 kDa蛋白质的纯化方案。在纯化过程中发现，患者血清中检测到的66 kDa蛋白质实际上是两种蛋白质，在十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳中均迁移至66 kDa处，在DE - 52色谱柱上洗脱于不同组分。这两种蛋白质分别被标记为牛角膜上皮抗原 - A和 - B（BCEA - A和BCEA - B）。对抗体结合的进一步研究表明，患者血清与这两种蛋白质中的一种或另一种结合，它们之间无交叉反应。将分离的BCEA - A和BCEA - B蛋白提取物与27例WG患者血清、10例变应性肉芽肿性血管炎（CSS）患者血清、31例类风湿关节炎（RA）患者血清以及血库中40例健康对照者血清进行免疫印迹分析。

结果

46%的WG患者血清含有针对66 kDa抗原之一的抗体，而健康对照者血清中均无66 kDa抗体（P < 10⁻⁵）。在WG组中，31%对BCEA - A呈阳性（与对照组相比，P = 0.0023），15%对BCEA - B呈阳性。与健康对照者相比，患有周边溃疡性角膜炎（PUK）的WG患者与抗BCEA - A抗体有显著相关性（50%，P < 10⁻⁶）。然而，在无眼部疾病的RA组中与BCEA - A有相关性（25%，P = 0.011），但在患有PUK的RA组中无相关性。CSS患者中抗BCEA - B抗体的频率显著增加（60%，P < 10⁻⁷）。