Adler G
Centrum Medyczne Kształcenia Podyplomowego, Zakład Biochemii, Warszawa.
Endokrynol Pol. 1992;43(1 Suppl):1-61.
The aim of this work was to characterise the eye muscle antigens reacting with autoantibodies from Graves ophthalmopathy patients to elucidate the function of these antibodies. As estimated by ELISA test antibodies of IgG class reacting with porcine microsomal membranes are present in about 25% of patients while of IgM class in about 15% of patients with Graves ophthalmopathy. Their presence do not correlate with ophthalmopathy index, neither they have relation to treatment. Anti eye muscle antibodies were present at some stages of the disease in three patients who develop ophthalmopathy, from the group of 26 patients treated during one year for hyperthyroidism. However, sporadically these antibodies were found also in about 20% of patients with Hashimoto disease, Lupus erythematosus or Scleroderma. Some of them cross react with antigens of skeletal muscle and liver. Eye muscle antigens reacting with patients antibodies are localised in plasma membranes and in membranes of smooth reticulum. Affinity purification of solubilised porcine eye muscle membrane proteins on a column with immunoglobulins from pooled serum of patients resulted in 23 fold purification of the antigen. The sensitivity of ELISA was not significantly increased by the use of affinity purified antigen, however some of previously negative ophthalmopathy sera gave positive reaction. Porcine eye muscle membrane proteins were separated by SDS PAGE and transferred to nitrocellulose. The reactions of electroblotted proteins with sera from patients with Graves ophthalmopathy and also sera from healthy controls shown very complex pattern. There was not a single antigen or antigens reacting only with antibodies present in sera of ophthalmopathy patients and not in controls. Patients sera reacted more often than control sera with an antigen of about 40 kDa. The reaction of sera from some patients with proteins about 100, 70 and 65 kDa were stronger than between these proteins and control sera. No changes in the pattern of reaction between antibodies and eye muscle antigens were noticed in serum of the same Graves' patient with or without ophthalmopathy during one year follow up and treatment, regardless of clinical course of the disease. When human eye muscle membrane fractions from tissue obtained during strabismus repair or at autopsy was used for immunoblotting, smaller number of proteins reacted with autoantibodies. Again there was no single antigen or antigens reacting with antibodies from sera of all Graves ophthalmopathy patients. Sera of some patients reacted with antigens about 50 kDa, not recognized by controls. The results of present study show, that the anti eye muscle antibodies are present in some of Graves ophthalmopathy patients.(ABSTRACT TRUNCATED AT 400 WORDS)
这项工作的目的是鉴定与格雷夫斯眼病患者自身抗体发生反应的眼肌抗原,以阐明这些抗体的功能。通过酶联免疫吸附测定(ELISA)测试估计,约25%的格雷夫斯眼病患者存在与猪微粒体膜发生反应的IgG类抗体,约15%的患者存在IgM类抗体。它们的存在与眼病指数无关,也与治疗无关。在一组因甲状腺功能亢进接受了一年治疗的26名患者中,有3名出现眼病的患者在疾病的某些阶段存在抗眼肌抗体。然而,在约20%的桥本氏病、红斑狼疮或硬皮病患者中也偶尔发现这些抗体。其中一些抗体与骨骼肌和肝脏的抗原发生交叉反应。与患者抗体发生反应的眼肌抗原定位于质膜和滑面内质网的膜上。用来自患者混合血清的免疫球蛋白在柱上对溶解的猪眼肌膜蛋白进行亲和纯化,使抗原得到了23倍的纯化。使用亲和纯化抗原后,ELISA的灵敏度没有显著提高,但一些先前呈阴性的眼病血清出现了阳性反应。猪眼肌膜蛋白通过十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS - PAGE)分离并转移到硝酸纤维素膜上。电印迹蛋白与格雷夫斯眼病患者血清以及健康对照血清的反应呈现出非常复杂的模式。没有单一的抗原或一组抗原仅与眼病患者血清中的抗体发生反应而不与对照血清中的抗体反应。患者血清比对照血清更常与一种约40 kDa的抗原发生反应。一些患者血清与约100、70和65 kDa的蛋白的反应比这些蛋白与对照血清的反应更强。在一名格雷夫斯病患者有或没有眼病的情况下,在一年的随访和治疗期间,无论疾病的临床进程如何,抗体与眼肌抗原之间的反应模式均未发现变化。当使用斜视修复手术中获取的组织或尸检时获得的人眼肌膜部分进行免疫印迹时,与自身抗体发生反应的蛋白数量较少。同样,没有单一的抗原或一组抗原与所有格雷夫斯眼病患者血清中的抗体发生反应。一些患者的血清与约50 kDa的抗原发生反应,而对照血清未识别该抗原。本研究结果表明,一些格雷夫斯眼病患者存在抗眼肌抗体。(摘要截选至400字)