Cederholm B, Wieslander J, Bygren P, Heinegård D
Department of Nephrology, University Hospital, Lund, Sweden.
Proc Natl Acad Sci U S A. 1988 Jul;85(13):4865-8. doi: 10.1073/pnas.85.13.4865.
IgA antibodies from patients with primary IgA nephropathy bind to collagens I, II, and IV. Here we show that this binding is mediated by the collagen-binding site of fibronectin, which occurs in the circulation in complex with IgA. No antibodies binding directly to collagen were identified. The complexes were isolated by affinity chromatography on gelatin-Sepharose and heparin-Sepharose, both with affinity for fibronectin, followed by adsorption to anti-human IgA immobilized on agarose gel. The presence of fibronectin and IgA antibodies in the isolated complexes is shown by enzyme-linked immunosorbent assay, gel electrophoresis, and electrophoretic transfer followed by immunostaining. The presence of an IgA-fibronectin complex in serum and the binding of this complex to collagen demonstrate the necessity of removing fibronectin from serum prior to identifying anti-collagen antibodies.
原发性IgA肾病患者的IgA抗体可与I型、II型和IV型胶原结合。在此我们发现,这种结合是由纤连蛋白的胶原结合位点介导的,纤连蛋白在循环中与IgA形成复合物。未鉴定出直接与胶原结合的抗体。通过在对纤连蛋白具有亲和力的明胶-琼脂糖凝胶和肝素-琼脂糖凝胶上进行亲和层析来分离复合物,随后吸附到固定在琼脂糖凝胶上的抗人IgA上。通过酶联免疫吸附测定、凝胶电泳以及电泳转移后进行免疫染色来显示分离出的复合物中存在纤连蛋白和IgA抗体。血清中存在IgA-纤连蛋白复合物以及该复合物与胶原的结合表明,在鉴定抗胶原抗体之前,有必要从血清中去除纤连蛋白。