Molecular composition of drusen and possible involvement of anti-retinal autoimmunity in two different forms of macular degeneration in cynomolgus monkey (Macaca fascicularis)
Molecular composition of drusen and possible involvement of anti-retinal autoimmunity in two different forms of macular degeneration in cynomolgus monkey (Macaca fascicularis)
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DOI：
10.1096/fj.04-3525fje
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发表时间：
2005-08-01
期刊：
影响因子：
4.8
通讯作者：
Iwata, T
中科院分区：
文献类型：
--
作者：
Umeda, S;Suzuki, MT;Iwata, T
我们以前曾报道过一个食蟹猴（Macaca fascicularis）家系，患有早发性黄斑变性，在出生后2年发生玻璃疣（1）。在这项研究中，分子组成的玻璃疣在猴子的影响与晚发性和早发性黄斑变性的特点。抗视网膜自身免疫在玻璃膜疣的形成和发病机制中的作用也进行了评价。对278只成年猴（平均年龄= 16.94岁）进行了眼底镜和组织学检查，以检查晚发性黄斑变性。通过免疫组织化学和/或使用液相色谱串联质谱（LC-MS/ MS）的直接蛋白质组分析来分析玻璃疣的分子组成。通过Western印迹技术在20只患病猴和10只年龄匹配的对照猴中筛选血清中的抗视网膜自身抗体。免疫原性分子进行了鉴定，2D电泳和LC-MS/MS。相对抗体滴度对每种抗原的ELISA测定42例受影响（晚发型）和41例正常猴的血清中。在90只（32%）接受检查的晚发型猴子中观察到黄斑区的黄白色斑点。组织学检查显示玻璃疣或视网膜色素上皮（RPE）细胞变性与色素异常有关。晚发和早发猴的玻璃疣均表现出载脂蛋白E、淀粉样蛋白P成分、补体成分C5、末端C5 b-9补体复合物、玻连蛋白和膜辅因子蛋白的免疫反应性。LC-MS/MS分析鉴定了60种蛋白质作为玻璃疣的成分，包括人类年龄相关性黄斑变性（AMD）玻璃疣中的许多常见组分，如膜联蛋白、晶体蛋白、免疫球蛋白和补体组分。一半的受影响的猴子有一个或多个自身抗体对38，40，50和60 kDa的视网膜蛋白。38和40 kDa的反应抗原被确定为膜联蛋白II和μ-晶体蛋白，分别。感染猴抗膜联蛋白II的相对抗体滴度显著高于对照组（P
We have previously reported a cynomolgus monkey (Macaca fascicularis) pedigree with early onset macular degeneration that develops drusen at 2 yr after birth (1). In this study, the molecular composition of drusen in monkeys affected with late onset and early onset macular degeneration was both characterized. Involvement of anti-retinalautoimmunity in the deposition of drusen and the pathogenesis of the disease was also evaluated. Funduscopic and histological examinations were performed on 278 adult monkeys (mean age = 16.94 yr) for late onset macular degeneration. The molecular composition of drusen was analyzed by immunohistochemistry and/or direct proteome analysis using liquid chromatography tandem mass spectroscopy (LC-MS/ MS). Anti-retinal autoantibodies in sera were screened in 20 affected and 10 age-matched control monkeys by Western blot techniques. Immunogenic molecules were identified by 2D electrophoresis and LC-MS/ MS. Relative antibody titer against each antigen was determined by ELISA in sera from 42 affected (late onset) and 41 normal monkeys. Yellowish-white spots in the macular region were observed in 90 (32%) of the late onset monkeys that were examined. Histological examination demonstrated that drusen or degenerative retinal pigment epithelium (RPE) cells were associated with the pigmentary abnormalities. Drusen in both late and early onset monkeys showed immunoreactivities for apolipoprotein E, amyloid P component, complement component C5, the terminal C5b-9 complement complex, vitronectin, and membrane cofactor protein. LC-MS/MS analyses identified 60 proteins as constituents of drusen, including a number of common components in drusen of human age-related macular degeneration (AMD), such as annexins, crystallins, immunoglobulins, and complement components. Half of the affected monkeys had single or multiple autoantibodies against 38, 40, 50, and 60 kDa retinal proteins. The reacting antigens of 38 and 40 kDa were identified as annexin II and mu-crystallin, respectively. Relative antibody titer against annexin II in affected monkeys was significantly higher than control animals (P