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使用肽阵列筛选系统性红斑狼疮自身抗原上的表位。

Screening epitopes on systemic lupus erythematosus autoantigens with a peptide array.

作者信息

Wang Lin, Hao Chenjun, Deng Yongqiu, Liu Yanbo, Hu Shiliang, Peng Yangang, He Manna, Fu Jinhu, Liu Ming, Chen Jia, Chen Xiaoming

机构信息

Department of Rheumatology, Shaoyang Central Hospital, 422000 Shaoyang, China.

Obstetrics and gynecology, Guangzhou Panyu Hexian Memorial Hospital, 511400 Guangzhou, China.

出版信息

Oncotarget. 2017 Sep 18;8(49):85559-85567. doi: 10.18632/oncotarget.20994. eCollection 2017 Oct 17.

Abstract

Systemic lupus erythematosus (SLE) is a common autoimmune disease. Many autoantibodies are closely associated with SLE. However, the specific epitopes recognized and bound by these autoantibodies are still unclear. This study screened the binding epitopes of SLE-related autoantibodies using a high-throughput screening method. Epitope prediction on 12 SLE-related autoantigens was performed using the Immune Epitope Database and Analysis Resource (IEDB) software. The predicted epitopes were synthesized into peptides and developed into a peptide array. Serum IgG from 50 SLE patients and 25 healthy controls was detected using the peptide array. The results were then validated using an enzyme-linked immunosorbent assay (ELISA). The diagnostic efficiency of each epitope was analyzed using a ROC curve. Seventy-three potential epitopes were screened for using the IEDB software after the epitopes on the 12 SLE-related autoantigens were analyzed. Peptide array screening revealed that the levels of the autoantibodies recognized and bound by 4 peptide antigens were significantly upregulated in the serum of SLE patients ( < 0.05). The ELISA results showed that the 4 antigens with significantly increased serum autoantibodies levels in SLE patients were acidic ribosomal phosphoprotein (P0)-4, acidic ribosomal phosphoprotein (P0)-11, DNA topoisomerase 1 (full length)-1, and U1-SnRNP 68/70 KDa-1 ( < 0.05), and the areas under the ROC curve for diagnosing SLE on the basis of these peptides were 0.91, 0.90, 0.93, and 0.91, respectively. Many autoantibodies specifically expressed in the serum of patients with SLE can be detected by specific peptide fragments and may be used as markers in clinical auxiliary diagnoses.

摘要

系统性红斑狼疮(SLE)是一种常见的自身免疫性疾病。许多自身抗体与SLE密切相关。然而,这些自身抗体识别和结合的特定表位仍不清楚。本研究采用高通量筛选方法筛选SLE相关自身抗体的结合表位。使用免疫表位数据库和分析资源(IEDB)软件对12种SLE相关自身抗原进行表位预测。将预测的表位合成肽段并制成肽阵列。使用该肽阵列检测50例SLE患者和25例健康对照者的血清IgG。然后使用酶联免疫吸附测定(ELISA)对结果进行验证。使用ROC曲线分析每个表位的诊断效率。在分析了12种SLE相关自身抗原的表位后,使用IEDB软件筛选出73个潜在表位。肽阵列筛选显示,4种肽抗原识别和结合的自身抗体水平在SLE患者血清中显著上调(<0.05)。ELISA结果显示,SLE患者血清中自身抗体水平显著升高的4种抗原分别为酸性核糖体磷蛋白(P0)-4、酸性核糖体磷蛋白(P0)-11、DNA拓扑异构酶1(全长)-1和U1-SnRNP 68/70 KDa-1(<0.05),基于这些肽段诊断SLE的ROC曲线下面积分别为0.91、0.90、0.93和0.91。SLE患者血清中特异性表达的许多自身抗体可被特定肽段检测到,并可能用作临床辅助诊断的标志物。

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