Yoshida Yoichi, Hiwasa Takaki, Machida Toshio, Kobayashi Eiichi, Mine Seiichiro, Matsushima Jun, Takiguchi Masaki, Iwadate Yasuo
Department of Neurological Surgery, Graduate School of Medicine, Chiba University.
Department of Biochemistry and Genetics, Graduate School of Medicine, Chiba University.
Neurol Med Chir (Tokyo). 2018 Jul 15;58(7):303-310. doi: 10.2176/nmc.ra.2018-0022. Epub 2018 May 31.
Recent clinical research has revealed a significant correlation between atherosclerosis, one of the primary etiologies of ischemic stroke, and the immune system. Assuming that "disease-specific autoantibodies are produced in the sera of patients with ischemic stroke," we investigated multiple arteriosclerosis-related antibodies using the serological identification of antigens by recombinant cDNA expression cloning (SEREX), an established method for identifying antigenic proteins. We either screened a human aortic endothelial cell cDNA library or conducted protein array screening using the sera from patients with ischemic stroke, such as carotid artery stenosis or transient ischemic attack (TIA). Next, we measured serum antibody levels using amplified luminescent proximity homogeneous assay-linked immunosorbent assay (AlphaLISA) in patient/healthy donor (HD) cohorts and identified several antigens, the antibody levels of which were significantly higher in patients with ischemic stroke than in HDs. This review introduced the method of identifying antigens by the SEREX and protein microarray and summarized antigenic proteins. In particular, it focused on anti-replication protein A2 antibody and anti-programmed cell death 11 antibody, which are significantly related to atherosclerotic plaque and ischemic brain tissue, respectively, and proposed the mechanism of elevated autoantibody levels against them. Furthermore, this review suggests a possibility of clinical application as an atherosclerotic disease diagnostic marker for TIA or cerebral infarction.
近期临床研究表明,缺血性中风的主要病因之一动脉粥样硬化与免疫系统之间存在显著关联。假设“缺血性中风患者血清中会产生疾病特异性自身抗体”,我们采用重组cDNA表达克隆血清学抗原鉴定法(SEREX)(一种用于鉴定抗原蛋白的成熟方法),对多种与动脉硬化相关的抗体进行了研究。我们要么筛选人主动脉内皮细胞cDNA文库,要么使用缺血性中风患者(如颈动脉狭窄或短暂性脑缺血发作(TIA)患者)的血清进行蛋白质阵列筛选。接下来,我们在患者/健康供体(HD)队列中使用放大发光邻近均相分析-酶联免疫吸附测定(AlphaLISA)测量血清抗体水平,并鉴定出几种抗原,缺血性中风患者体内这些抗原的抗体水平显著高于健康供体。本综述介绍了通过SEREX和蛋白质微阵列鉴定抗原的方法,并总结了抗原蛋白。特别关注了分别与动脉粥样硬化斑块和缺血性脑组织显著相关的抗复制蛋白A2抗体和抗程序性细胞死亡11抗体,并提出了针对它们的自身抗体水平升高的机制。此外,本综述提出了将其作为TIA或脑梗死的动脉粥样硬化疾病诊断标志物进行临床应用的可能性。