Peene I, Meheus L, De Keyser S, Humbel R, Veys E M, De Keyser F
Department of Rheumatology, University Hospital Gent, Belgium Innogenetics, Gent, Belgium Centre Hospitalier Luxembourg, Luxembourg.
Ann Rheum Dis. 2002 Oct;61(10):929-33. doi: 10.1136/ard.61.10.929.
To determine whether anti-Ro52 is an independent serum marker in connective tissue disease.
Over a two year period, 1727 consecutive antinuclear antibody (ANA) positive serum samples were analysed in parallel by double immunodiffusion with thymus/spleen nuclear extract and by line immunoassay with recombinant Ro52, recombinant La/SSB, and natural Ro60. Sera that were only reactive towards Ro52 were further analysed by a variety of additional anti-SSA/Ro detection methods and by specific anti-Ro52 and anti-Ro60 assays. Natural purified SSA/Ro was analysed by immunoblot and protein sequencing.
Analysis of natural purified SSA/Ro (Immunovision, Springdale, AR) showed only Ro60 and no immunoreactive Ro52. Consequently, assays based on this substrate only identify sera with anti-Ro60 reactivity. Twenty serum samples showed anti-Ro52 without anti-Ro60 and anti-SSB/La on line immunoassay. By additional testing, 2/20 sera were found positive for anti-Ro60 reactivity. The remaining 18 sera were not identified by any of the classical anti-SSA/Ro assays and were considered to be reactive only with Ro52 and not with Ro60. This anti-Ro52 reactivity was confirmed by natural and recombinant Ro52 in 16/18 cases. 12/18 sera corresponded to connective tissue diseases.
Anti-Ro52 positive sera without any evidence of anti-Ro60 and anti-La/SSB reactivity can be considered as an independent group that is systematically missed by classical anti-SSA/Ro detection methods owing to a bias towards anti-Ro60 reactivity. The anti-Ro52 sera are precipitin negative, not retrieved by SSA/Ro enzyme linked immunosorbent assays (ELISAs) based on natural SSA/Ro, and show no specific ANA fluorescence staining pattern. These findings together with the clinical data indicate that anti-Ro52 should be considered as an additional and independent serum marker.
确定抗Ro52是否为结缔组织病中的一种独立血清标志物。
在两年时间里，连续对1727份抗核抗体（ANA）阳性血清样本进行分析，采用胸腺/脾脏核提取物双向免疫扩散法以及重组Ro52、重组La/SSB和天然Ro60的线性免疫分析法进行平行检测。仅对Ro52有反应的血清通过多种额外的抗SSA/Ro检测方法以及特异性抗Ro52和抗Ro60检测进行进一步分析。对天然纯化的SSA/Ro进行免疫印迹和蛋白质测序分析。
对天然纯化的SSA/Ro（Immunovision，Springdale，阿肯色州）分析显示只有Ro60，没有免疫反应性的Ro52。因此，基于该底物的检测仅能识别出具有抗Ro60反应性的血清。20份血清样本在线性免疫分析中显示抗Ro52阳性，且无抗Ro60和抗SSB/La反应性。通过额外检测，发现2/20的血清抗Ro60反应性呈阳性。其余18份血清未被任何经典抗SSA/Ro检测方法识别，被认为仅与Ro52反应，而不与Ro60反应。16/18例中天然和重组Ro52证实了这种抗Ro52反应性。12/1十八份血清对应结缔组织病。
无任何抗Ro60和抗La/SSB反应性证据的抗Ro52阳性血清可被视为一个独立的组，由于偏向抗Ro60反应性，经典抗SSA/Ro检测方法系统性地遗漏了该组。抗Ro52血清沉淀素阴性，基于天然SSA/Ro的SSA/Ro酶联免疫吸附测定（ELISA）无法检测到，且无特异性ANA荧光染色模式。这些发现与临床数据表明，抗Ro52应被视为一种额外的独立血清标志物。
