Department of Rheumatology and Clinical Immunology, Faculty of Medicine, School of Health Sciences, University of Thessaly, Larissa, Greece.
Department of Immunology-Histocompatibility, Evangelismos General Hospital, Athens, Greece.
Front Immunol. 2018 Dec 7;9:2835. doi: 10.3389/fimmu.2018.02835. eCollection 2018.
Epitope mapping of anti-Ro52 antibodies (Abs) has been extensively studied in patients with Sjögren's syndrome (SjS) and systemic lupus erythematosus (SLE). Comprehensive epitope mapping in systemic sclerosis (SSc), where anti-Ro52 antibodies are also frequently detected, has not been performed. The aim of the present study was to fully characterize Ro52 epitopes in anti-Ro52-positive SSc using Ro52 fragments spanning the full antigen. Further analysis was made according to anti-Ro60 status. Epitope mapping was performed in 43 anti-Ro52-positive SSc patients. Seventy eight anti-Ro52-positive pathological controls, including 20 patients with SjS, 28 patients with SLE, 15 patients with dermatomyositis (DM), and 15 patients with primary biliary cholangitis (PBC), and 20 anti-Ro52-negative healthy individuals as normal controls were also tested. Five recombinant Ro52 fragments [Ro52-1 (aa 1-127), Ro52-2 (aa 125-268), Ro52-3 (aa 268-475), Ro52-4 (aa 57-180), and Ro52-5 (aa 181-320) were used to test reactivity by line-immunoassay and ELISA. Anti-Ro60 reactivity was tested by ELISA. All anti-Ro52 positive sera reacted with Ro52-2; none recognized Ro52-3. Antibodies against Ro52-1 were less frequently found in SSc than in SjS/SLE (11.6 vs. 41.7%, = 0.001); and antibodies against Ro52-4 were less frequently found in SSc than in SjS/SLE (27.9 vs. 50%, = 0.03). In SSc patients, reactivity against Ro52-1 was more frequent in anti-Ro52+/anti-Ro60+ than in anti-Ro52+/anti-Ro60-patients (33.3 vs. 0%, = 0.003). In this comprehensive analysis of Ro52 epitope mapping in SSc, the coiled coil domain remains the predominant epitope on Ro52. Contrary to SjS and SLE, patients with SSc fail to identify epitopic regions within the N-terminus of the protein, especially if they lack con-current anti-Ro60 reactivity.
抗 Ro52 抗体（Abs）的表位作图在干燥综合征（SjS）和系统性红斑狼疮（SLE）患者中已得到广泛研究。在系统性硬化症（SSc）中也经常检测到抗 Ro52 抗体，但尚未进行全面的表位作图。本研究的目的是使用跨越全长抗原的 Ro52 片段，全面描述抗 Ro52 阳性 SSc 中的 Ro52 表位。根据抗 Ro60 状态进一步分析。对 43 例抗 Ro52 阳性 SSc 患者进行了表位作图。还测试了 78 例抗 Ro52 阳性病理对照，包括 20 例 SjS 患者、28 例 SLE 患者、15 例皮肌炎（DM）患者和 15 例原发性胆汁性胆管炎（PBC）患者，以及 20 例抗 Ro52 阴性健康个体作为正常对照。使用 5 个重组 Ro52 片段[Ro52-1（aa1-127）、Ro52-2（aa125-268）、Ro52-3（aa268-475）、Ro52-4（aa57-180）和 Ro52-5（aa181-320）]通过线免疫测定和 ELISA 测试反应性。通过 ELISA 测试抗 Ro60 反应性。所有抗 Ro52 阳性血清均与 Ro52-2 反应；没有一种识别 Ro52-3。与 SjS/SLE 相比，SSc 中抗 Ro52-1 的抗体较少（11.6%比 41.7%，=0.001）；并且与 SjS/SLE 相比，SSc 中抗 Ro52-4 的抗体较少（27.9%比 50%，=0.03）。在 SSc 患者中，与抗 Ro52+/抗 Ro60+患者相比，抗 Ro52+/抗 Ro60-患者对 Ro52-1 的反应性更频繁（33.3%比 0%，=0.003）。在 SSc 中对 Ro52 表位作图的全面分析中，卷曲螺旋结构域仍然是 Ro52 的主要表位。与 SjS 和 SLE 不同，如果 SSc 患者缺乏同时存在的抗 Ro60 反应性，则无法识别蛋白 N 端的表位区域。
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