Identification of a target antigen in human anti-tubular basement membrane nephritis.
Identification of a target antigen in human anti-tubular basement membrane nephritis.
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人抗肾小管基底膜肾炎靶抗原的鉴定。
DOI：
10.1038/ki.1987.69
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发表时间：
1987
影响因子：
19.6
通讯作者：
Butkowski,RJ
中科院分区：
文献类型：
--
作者：
Fliger,FD;Wieslander,J;Brentjens,JR;Andres,GA;Butkowski,RJ
人抗肾小管基底膜肾炎靶抗原的鉴定。两名原发性抗肾小管基底膜介导的肾小管间质性肾炎患者（一名为肾移植受者，另一名为自发性抗肾小管基底膜疾病）的血清分析了其自身抗体的特异性。两种血清都有循环抗体，通过ELISA与几个物种的肾小管基底膜提取物反应，但与肾小球基底膜提取物反应不明显。反应性抗原用6 M盐酸胍、6 M尿素、还原和烷基化以及十二烷基硫酸钠溶解。用胶原酶消化基底膜，从膜中释放相对少量的抗原，胰蛋白酶和胃蛋白酶破坏其抗原性。通过凝胶过滤和蛋白质印迹的免疫覆盖分析，表征抗原活性的大小分布。总之，结果表明，两种血清的主要反应性均针对肾小管基底膜特有的Mr58，000组分。通过免疫覆盖分析检测到对肾小球和肾小管基底膜共同的高分子量组分的轻微反应性。本研究确定了一种抗原，参与人抗肾小管基底膜介导的肾小管间质性肾炎，并证明了使用变性提取蛋白水解方法制备抗原的优势。
Identification of a target antigen in human anti-tubular basement membrane nephritis. Sera from two patients with primary anti-tubular–basement–membrane–mediated tubulointerstitial nephritis, one a renal allograft recipient and the other with spontaneous anti-tubular–basement–membrane disease, were analyzed for the specificity of their autoantibodies. Both sera had circulating antibodies that reacted by ELISA with extracts of tubular basement membrane from several species, but failed to react significantly with extracts of glomerular basement membrane. Reactive antigen was solubilized with 6Mguanidine-HCl, 6Murea, with reduction and alkylation, and with sodium dodecylsulfate. Digestion of the basement membrane with collagenase released relatively small quantities of antigen from the membrane, and trypsin and pepsin destroyed its antigenicity. The antigenic activity was characterized with respect to its size distribution by gel filtration and by immuno-overlay analysis of protein blots. Collectively, the results indicate that the major reactivity of both sera is directed towards a Mr58,000 component that is unique to the tubular basement membrane. Minor reactivities toward high molecular weight components common to both glomerular and tubular basement membranes were detected by immuno-overlay analysis. This study identifies an antigen that is involved in human anti-tubular–basement–membrane–mediated tubulointerstitial nephritis, and demonstrates an advantage of the use of denaturing extraction over proteolytic methods to prepare the antigen.