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首页> 外文期刊>Arthritis research & therapy. >A novel DNA/histone H4 peptide complex detects autoantibodies in systemic lupus erythematosus sera
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A novel DNA/histone H4 peptide complex detects autoantibodies in systemic lupus erythematosus sera

机译：一种新型的DNA /组蛋白H4肽复合物可检测系统性红斑狼疮血清中的自身抗体

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Background The detection of anti-dsDNA antibodies is critical for the diagnosis and follow-up of systemic lupus erythematosus (SLE) patients. The presently available assays are characterized by a non-optimal specificity (solid phase assays) or sensitivity ( Crithidia Luciliae immunofluorescence test (CLIFT)). To overcome the limits of CLIFT and solid phase chromatin assays, we explored the diagnostic potential of an assay based on plasmid DNA containing a highly bent fragment of 211?bp from Crithidia Luciliae minicircles, complexed with histone peptides. Methods Electrically neutral complexes of PK201/CAT plasmid (PK) DNA and histone 4 (H4) peptides were evaluated by electromobility shift assay. Complexes of H4 peptides and PK were absorbed to the solid phase to detect specific immunoglobulin G (IgG) in sera. Sera from 109 SLE patients, 100 normal healthy subjects, and 169 disease controls were tested. Results H4(14-34) containing the consensus sequence for DNA binding interacts with PK, retarding its migration. H4(14-34)/PK complexes were used to test sera by ELISA. Anti-H4-PK antibodies were detected in 56?% of SLE sera (more frequently in patients with skin or joint involvement) versus 5.9?% in disease controls; inhibition assays show that sera react with epitopes present on DNA or on the complex, not on the peptide. Antibody titer is correlated with European Consensus Lupus Activity Measurement (ECLAM) score and anti-complement component 1q (C1q) antibodies, negatively with C3 levels. Anti-H4-PK antibodies compared with CLIFT and solid phase dsDNA assays display moderate concordance. Conclusions The H4/PK assay is a simple and reliable test which is useful for the differential diagnosis and evaluation of disease activity in SLE patients.
机译：背景技术抗dsDNA抗体的检测对于系统性红斑狼疮（SLE）患者的诊断和随访至关重要。当前可用的测定法的特征在于非最佳的特异性（固相测定法）或敏感性（露西丽氏菌免疫荧光试验（CLIFT））。为了克服CLIFT和固相染色质检测的局限性，我们探索了基于质粒DNA的检测方法的诊断潜力，该质粒DNA包含来自Crithidia Luciliae minicircles的高度弯曲的211？bp片段，并与组蛋白肽复合。方法采用电动迁移法测定PK201 / CAT质粒（PK）DNA和组蛋白4（H4）肽的电中性复合物。 H4肽和PK的复合物被吸收到固相，以检测血清中的特异性免疫球蛋白G（IgG）。测试了109名SLE患者，100名正常健康受试者和169名疾病对照的血清。结果H4（14-34）包含DNA结合的共有序列与PK相互作用，阻碍了它的迁移。 H4（14-34）/ PK复合物用于通过ELISA检测血清。在SLE血清中检出抗H4-PK抗体的比例为56％（在皮肤或关节受累的患者中更为频繁），而在疾病对照中则为5.9％。抑制试验表明，血清与DNA或复合物上而不是肽上的表位反应。抗体效价与欧洲共识狼疮活性测量（ECLAM）评分和抗补体成分1q（C1q）抗体相关，与C3水平呈负相关。与CLIFT和固相dsDNA分析相比，抗H4-PK抗体显示出中等程度的一致性。结论H4 / PK检测是一种简单可靠的检测方法，可用于SLE患者的鉴别诊断和疾病活动性评估。

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