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系统性红斑狼疮患者血浆中大分子DNA和RNA的抗体结合情况。

Antibody binding of macromolecular DNA and RNA in the plasma of SLE patients.

作者信息

Krapf F, Herrmann M, Leitmann W, Kalden J R

机构信息

Institute for Clinical Immunology and Rheumatology, University of Erlangen-Nünberg, FRG.

出版信息

Clin Exp Immunol. 1989 Mar;75(3):336-42.

Abstract

Plasmapheresis fluids from 20 patients with clinically active SLE, from three patients with Waldenstrom's disease, from three patients with rheumatoid arthritis, two patients with myasthenia gravis and other diseases including active systemic disorders were precipitated using polyethylene glycol 6000 (PEG). By applying ethidium bromide staining, plasma nucleic acids (PNA) could be demonstrated in PEG-precipitates of SLE patients exclusively. Purified immunoglobulins of SLE plasma precipitates were shown to form antigen-antibody complexes with PNA as demonstrated by electronmicroscopy. Further characterization of PNA by agarose gel electrophoresis revealed a molecular weight up to 20 kbp. Cesium chloride buoyant density gradients showed non-homogeneous molecules, excluding pure microbial origin. In spite of RNase digestion, the PNA contained RNA with 30-70% riboguanosine as shown by nucleoside analysis. The high amount of guanosine-rich RNA was further supported by similarities between PNA and polyriboguanylic acid in hyperchrome shifting due to thermic denaturation. HPLC analysis showed a molecular weight of ribonucleic acids of more than 60 b thus excluding mere oligonucleotides. In contrast to B-type dsDNA, PNA from SLE patients were immunogenic. Antibodies against PNA could be induced in rabbits by subcutaneous injection. The antisera thus obtained showed crossreactivity with polyriboguanylic acid and dsDNA preparations.

摘要

使用聚乙二醇6000(PEG)对20例临床活动期系统性红斑狼疮(SLE)患者、3例华氏巨球蛋白血症患者、3例类风湿性关节炎患者、2例重症肌无力患者以及包括活动性全身性疾病在内的其他疾病患者的血浆置换液进行沉淀。通过应用溴化乙锭染色,仅在SLE患者的PEG沉淀物中可证实存在血浆核酸(PNA)。电子显微镜显示,SLE血浆沉淀物的纯化免疫球蛋白与PNA形成抗原-抗体复合物。通过琼脂糖凝胶电泳对PNA进行进一步表征,结果显示其分子量高达20千碱基对(kbp)。氯化铯浮力密度梯度显示分子不均匀,排除了纯粹的微生物来源。核苷分析表明,尽管经过核糖核酸酶消化,PNA仍含有30%-70%为核糖鸟苷的RNA。PNA与聚核糖鸟苷酸在热变性导致的增色移位方面具有相似性,进一步支持了富含鸟苷的RNA含量较高这一结论。高效液相色谱(HPLC)分析显示核糖核酸的分子量超过60个碱基,因此排除了单纯的寡核苷酸。与B型双链DNA(dsDNA)不同,SLE患者的PNA具有免疫原性。通过皮下注射可在兔体内诱导产生针对PNA的抗体。由此获得的抗血清与聚核糖鸟苷酸和dsDNA制剂显示出交叉反应性。

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