Alpha Actinin is Specifically Recognized by Multiple Sclerosis Autoantibodies Isolated Using an N-Glucosylated Peptide Epitope
Alpha Actinin is Specifically Recognized by Multiple Sclerosis Autoantibodies Isolated Using an N-Glucosylated Peptide Epitope
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DOI：
10.1074/mcp.m112.017087
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发表时间：
2013-02-01
影响因子：
7
通讯作者：
Rovero, Paolo
中科院分区：
文献类型：
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作者：
Pandey, Shashank;Dioni, Ilaria;Rovero, Paolo
近年来，复杂的方法鉴定出了与多发性硬化症（MuS）相关的新型自身抗原，例如：G.神经成束蛋白、接触蛋白、CN β和其他T细胞受体膜锚定蛋白。这些推定的抗原，虽然不同于传统的髓鞘衍生物，概念上是基于实验性自身免疫性脑脊髓炎的动物模型。在这份报告中，我们描述了推定的抗原识别的基础上，从MuS患者血清中分离的自身抗体。在该实验室以前的工作中，我们已经证明了一种名为CSF 114（Glc）的肽探针特异性识别MuS患者亚组中的血清自身抗体，该亚组代表了30%的患者人群。通过CSF 114（Glc）亲和层析从MuS患者血清（6）纯化的自身抗体检测到存在于大鼠脑中的三个免疫反应性蛋白带。蛋白质组学分析的免疫反应条带，涉及MALDI和MS/MS技术，揭示了四种蛋白质的存在下区分其质量：α fodrin，α辅肌动蛋白1，肌酸激酶，和CNGINA. These大鼠脑蛋白的免疫反应谱进行了比较，与商业上可获得的标准蛋白质的挑战对CSF 114（葡萄糖）纯化的MuS自身抗体，或单克隆抗体。通过以下程序进一步区分大鼠脑蛋白：而单克隆抗体识别所有大鼠脑蛋白，分离的MuS特异性抗体仅识别α辅肌动蛋白1作为推定抗原。事实上，α辅肌动蛋白1对所有检测的MuS患者血清显示出强烈的免疫反应，而其他三条带不能始终检测到。因此，α辅肌动蛋白1，一种与炎症/退行性自身免疫性疾病（狼疮肾炎和自身免疫性肝炎）有关的细胞骨架蛋白，可能被视为一种新的MuS自身抗原，可能是该疾病典型的炎症/退行性过程的原型生物标志物。Molecular & Cellular Proteomics 12：10.1074/mcp. M112.017087，277-282，2013.
Sophisticated approaches have recently led to the identification of novel autoantigens associated with Multiple Sclerosis (MuS), e. g. neurofascin, contactin, CNPase, and other T-cell receptor membrane anchored proteins. These putative antigens, although differing from the conventional myelin derivatives, are conceptually based on an animal model of experimental autoimmune encephalomyelitis. In this report we describe the identification of putative antigens based on their recognition by autoantibodies isolated from MuS patient serum. In a previous work from this laboratory we have shown that a peptide probe, named CSF114(Glc), specifically identifies serum autoantibodies in a subset of MuS patients, representing similar to 30% of the patient population. The autoantibodies, purified from MuS patients' sera (six), through CSF114(Glc) affinity chromatography, detected three immunoreactive protein bands present in the rat brain. Proteomic analysis of the immunoreactive bands, involving MALDI and MS/MS techniques, revealed the presence of four proteins distinguishable by their mass: alpha fodrin, alpha actinin 1, creatine kinase, and CNPase.The immunoreactive profile of these rat brain proteins was compared with that of commercially available standard proteins by challenging against either CSF114(Glc) purified MuS autoantibodies, or monoclonal antibodies. Further discrimination among the rat brain proteins was provided by the following procedure: whereas monoclonal antibodies recognized all rat brain proteins, isolated MuS specific antibodies recognize only alpha actinin 1 as a putative antigen. In fact, alpha actinin 1 displayed a robust immunoreactive response against all MuS patients' sera examined, whereas the other three bands were not consistently detectable. Thus, alpha actinin 1, a cytoskeleton protein implicated in inflammatory/degenerative autoimmune diseases (lupus nephritis and autoimmune hepatitis) might be regarded as a novel MuS autoantigen, perhaps a prototypic biomarker for the inflammatory/degenerative process typical of the disease. Molecular & Cellular Proteomics 12: 10.1074/mcp.M112.017087, 277-282, 2013.