Gottsch J D, Liu S H, Minkovitz J B, Goodman D F, Srinivasan M, Stark W J
Wilmer Eye Institute, Johns Hopkins Hospital, Baltimore, MD 21205, USA.
Invest Ophthalmol Vis Sci. 1995 Jul;36(8):1541-7.
To purify and characterize a cornea-associated antigen (CO-Ag) and to determine antibody levels to CO-Ag in patients with Mooren's ulcer.
Standard ion exchange and gel filtration chromatographies were used to isolate and purify CO-Ag from crude bovine stromal extracts. The serum of a patient with Mooren's ulcer, containing a high level of antibodies directed against CO-Ag, was used to monitor isolation procedures. Using this newly purified CO-Ag, an enzyme-linked immunoabsorbent assay was used to detect the presence of antibodies to CO-Ag in the sera of other patients with Mooren's ulcer.
CO-Ag was purified to apparent homogeneity from bovine corneal stromal extracts by a series of ion exchange chromatographies and gel filtration. Polyacrylamide gel electrophoresis showed that CO-Ag was a tetramer with a molecular weight of 30,000 d that may dissociate under denaturing conditions into a monomer of 7000 d. Strong indirect immunofluorescent staining was demonstrated of the stroma by guinea pig anti-CO-Ag antibody. A statistically significant difference in the level of specific antibodies to CO-Ag between patients with Mooren's ulcer and controls was found (P < 0.001). The antibody level was elevated in patients with Mooren's ulcer (mean antibody level, 0.58 +/- 0.13) compared with the controls (mean antibody level, 0.22 +/- 0.04).
These results suggest that an autoantigen exists in the corneal stroma that reacts with serum antibodies from patients with Mooren's ulcer. The availability of a purified corneal antigen could facilitate the diagnosis and define the pathogenetic mechanisms in Mooren's ulcer.
纯化并鉴定一种角膜相关抗原(CO-Ag),并测定蚕食性角膜溃疡患者体内针对CO-Ag的抗体水平。
采用标准离子交换色谱法和凝胶过滤色谱法从牛角膜基质粗提物中分离纯化CO-Ag。一位患有蚕食性角膜溃疡且体内含有高水平抗CO-Ag抗体的患者血清被用于监测分离过程。利用这种新纯化的CO-Ag,采用酶联免疫吸附测定法检测其他蚕食性角膜溃疡患者血清中抗CO-Ag抗体的存在情况。
通过一系列离子交换色谱法和凝胶过滤法,从牛角膜基质提取物中纯化得到了外观均一的CO-Ag。聚丙烯酰胺凝胶电泳显示,CO-Ag是一种分子量为30,000 d的四聚体,在变性条件下可能解离为7000 d的单体。豚鼠抗CO-Ag抗体对角膜基质呈现出强烈的间接免疫荧光染色。发现蚕食性角膜溃疡患者与对照组之间针对CO-Ag的特异性抗体水平存在统计学显著差异(P < 0.001)。与对照组(平均抗体水平为0.22 ± 0.04)相比,蚕食性角膜溃疡患者的抗体水平升高(平均抗体水平为0.58 ± 0.13)。
这些结果表明,角膜基质中存在一种自身抗原,它能与蚕食性角膜溃疡患者的血清抗体发生反应。纯化的角膜抗原的可得性有助于蚕食性角膜溃疡的诊断并明确其发病机制。