Broekhuyse R M, Kuhlmann E D
Institute of Ophthalmology, University of Nijmegen, The Netherlands.
Invest Ophthalmol Vis Sci. 1993 Mar;34(3):698-700.
The purpose of this study was to develop a rapid procedure for the preparation of melanin with a specific, highly uveitogenic activity.
A crude melanosome fraction was isolated from bovine choroids (containing remnants of adhering retinal pigment epithelium). The fraction was extracted with hot 2% sodium dodecyl sulfate, and Lewis rats were immunized with the purified melanin, using pertussis toxin as coadjuvant.
The purified melanin was free from pathogenic photoreceptor antigens and other accompanying or adsorbed proteins. It was able to evoke severe, acute, anterior uveitis with the typical characteristics of experimental autoimmune anterior uveitis (EAAU; without retinitis or pinealitis), even at the level of 1 micrograms melanin protein.
The rapidly prepared ocular melanin exhibits the same qualities as purified choroidal or retinal pigment melanins obtained by much more laborious procedures (which also deliver other subcellular fractions for investigation). It is suitable for the study of the immunopathogenesis of EAAU, which is a new model for human acute anterior uveitis.
本研究的目的是开发一种快速制备具有特定、高度致葡萄膜炎活性的黑色素的方法。
从牛脉络膜（含有附着的视网膜色素上皮残余物）中分离出粗制的黑素体部分。用热的2%十二烷基硫酸钠提取该部分，并用百日咳毒素作为佐剂，用纯化的黑色素对Lewis大鼠进行免疫。
纯化的黑色素不含致病性光感受器抗原及其他伴随或吸附的蛋白质。即使在1微克黑色素蛋白的水平，它也能够引发严重的急性前葡萄膜炎，具有实验性自身免疫性前葡萄膜炎（EAAU；无视网膜炎或松果体炎）的典型特征。
快速制备的眼黑色素具有与通过更费力的方法获得的纯化脉络膜或视网膜色素黑色素相同的性质（这些方法也提供其他亚细胞部分用于研究）。它适用于研究EAAU的免疫发病机制，EAAU是人类急性前葡萄膜炎的一种新模型。
