Department of Surgery, Medical University of Vienna, A-1090 Vienna, Austria.
J Proteome Res. 2010 Feb 5;9(2):1041-9. doi: 10.1021/pr900930d.
The only treatment of end-stage renal disease patients undergoing chronic dialysis is kidney transplantation. However, about half of graft recipients encounter organ loss within ten years after renal transplantation. There is emerging evidence that the presence of alloreactive antibodies against non-HLA antigens in the serum of the recipient prior transplantation is associated with higher incidence of chronic rejection. However, the molecular identity of these antigens is largely unknown. To determine the most common non-HLA antigens, we tested lymphocytic extracts from 20 healthy volunteers with sera of 28 patients on the transplantation waiting list by Western blotting. There was a group of five proteins that was recognized by most sera. Using patient's own lymphocytes revealed that autoimmunity plays a minor role in this recognition. Two-dimensional Western blotting experiments followed by mass spectrometry identified the antigens as tubulin beta chain, vimentin, lamin-B1, and Rho GDP-dissociation inhibitor 2. A detailed analysis of vimentin expression revealed that the antigenic 60 kDa isoform is underrepresented in patient's lymphocytes in comparison to those of healthy volunteers. The study revealed that preformed alloreactive antibodies are directed against a small number of specific protein isoforms. Our findings could provide a basis for future improvement of donor-recipient matching.
终末期肾病患者接受慢性透析的唯一治疗方法是肾移植。然而，大约一半的移植物受者在肾移植后十年内会遭遇器官丧失。有新的证据表明，移植前受者血清中存在针对非 HLA 抗原的同种反应性抗体与慢性排斥反应的发生率较高有关。然而，这些抗原的分子身份在很大程度上是未知的。为了确定最常见的非 HLA 抗原，我们通过 Western blot 法检测了 20 名健康志愿者的淋巴细胞提取物和 28 名移植等待名单上患者的血清。有一组 5 种蛋白质被大多数血清识别。使用患者自身的淋巴细胞表明，自身免疫在这种识别中起次要作用。二维 Western blot 实验和质谱分析鉴定出的抗原为微管蛋白 β 链、波形蛋白、核纤层蛋白-B1 和 Rho GDP 解离抑制剂 2。对波形蛋白表达的详细分析表明，与健康志愿者相比，患者淋巴细胞中抗原性 60 kDa 同工型的表达不足。该研究表明，预先形成的同种反应性抗体针对少数特定的蛋白质同工型。我们的发现可以为未来改进供体-受者匹配提供基础。
