Immunoproteomic analysis of potential serum biomarker candidates in human glaucoma.
Immunoproteomic analysis of potential serum biomarker candidates in human glaucoma.
复制标题
DOI：
10.1167/iovs.12-10076
复制
发表时间：
2012-12
影响因子：
4.4
通讯作者：
G. Tezel;Ivey L. Thornton;M. Tong;C. Luo;Xiangjun Yang;Jian Cai;D. Powell;J. Soltau;J. Liebmann-J.
中科院分区：
文献类型：
--
作者：
G. Tezel;Ivey L. Thornton;M. Tong;C. Luo;Xiangjun Yang;Jian Cai;D. Powell;J. Soltau;J. Liebmann-J.
支持免疫系统参与青光眼的证据包括视网膜和视神经蛋白的血清抗体滴度增加，尽管它们的致病重要性尚不清楚。本研究使用基于抗体的蛋白质组学方法，旨在鉴定作为青光眼候选生物标志物的疾病相关抗原。方法收集111例原发性开角型青光眼患者和49例年龄匹配的健康对照者的血清样本。对于抗原的高通量表征，从五个随机选择的昏迷样品中洗脱血清IgG，并通过线性离子阱质谱法（LC-MS/MS）进行分析。然后通过特异性ELISA在整个样品池（包括糖尿病视网膜病变的另外对照组）中测量所选生物标志物候选物的血清滴度。结果IgG洗脱液的LC-MS/MS分析显示了一组复杂的蛋白质，包括仅在昏迷样品中可检测到的蛋白质。有趣的是，这些抗原中的许多抗原对应于先前在青光眼供体中鉴定的上调的视网膜蛋白（或表现出增加的甲硫氨酸氧化）。此外，额外的分析检测到患者血清对青光眼视网膜蛋白（或氧化应激细胞培养蛋白）的免疫反应性更高，从而表明疾病相关蛋白修饰在自身抗体产生/反应性中的重要性。作为选择初始生物标志物候选物的缩小策略，我们确定了与已知在青光眼中上调的视网膜蛋白重叠的血清蛋白。所选的10个候选物中的4个（AIF、环AMP反应元件结合蛋白、肝配蛋白A型受体和亨廷顿蛋白）在脑水肿血清中表现出较高的ELISA滴度。结论：青光眼免疫蛋白质组学研究鉴定的血清蛋白质可能代表病变组织相关抗原，可作为青光眼的候选生物标志物。
PURPOSE Evidence supporting the immune system involvement in glaucoma includes increased titers of serum antibodies to retina and optic nerve proteins, although their pathogenic importance remains unclear. This study using an antibody-based proteomics approach aimed to identify disease-related antigens as candidate biomarkers of glaucoma. METHODS Serum samples were collected from 111 patients with primary open-angle glaucoma and an age-matched control group of 49 healthy subjects without glaucoma. For high-throughput characterization of antigens, serum IgG was eluted from five randomly selected glaucomatous samples and analyzed by linear ion trap mass spectrometry (LC-MS/MS). Serum titers of selected biomarker candidates were then measured by specific ELISAs in the whole sample pool (including an additional control group of diabetic retinopathy). RESULTS LC-MS/MS analysis of IgG elutes revealed a complex panel of proteins, including those detectable only in glaucomatous samples. Interestingly, many of these antigens corresponded to upregulated retinal proteins previously identified in glaucomatous donors (or that exhibited increased methionine oxidation). Moreover, additional analysis detected a greater immunoreactivity of the patient sera to glaucomatous retinal proteins (or to oxidatively stressed cell culture proteins), thereby suggesting the importance of disease-related protein modifications in autoantibody production/reactivity. As a narrowing-down strategy for selection of initial biomarker candidates, we determined the serum proteins overlapping with the retinal proteins known to be up-regulated in glaucoma. Four of the selected 10 candidates (AIF, cyclic AMP-responsive element binding protein, ephrin type-A receptor, and huntingtin) exhibited higher ELISA titers in the glaucomatous sera. CONCLUSIONS A number of serum proteins identified by this immunoproteomic study of human glaucoma may represent diseased tissue-related antigens and serve as candidate biomarkers of glaucoma.