Weber M, Köhler H, Manns M, Baum H P, Meyer zum Büschenfelde K H
Clin Exp Immunol. 1987 Feb;67(2):262-9.
Collagenase-digests of human glomerular (GBM), alveolar (ABM), and placenta basement membranes (PBM) were separated by gel filtration columns and the pools rich in Goodpasture antigens (GP) were identified by an antibody inhibition-ELISA. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) followed by immunoblotting on nitrocellulose membranes was performed with each basement membrane preparation. Sera from patients with florid GP-syndrome and antibodies to glomerular basement membrane (anti-GBM antibodies) were incubated with nitrocellulose strips of GBM, ABM, and PBM. Immunoperoxidase staining revealed reactivity with target antigens of 24, 26, 44, and 50 kD in the GBM and of 44 and 50 kD in the ABM and PBM, respectively. No corresponding reactivity was observed using convalescent GP-sera, sera from patients with other immunological diseases or sera from healthy blood donors. The antigens were sensitive to reduction. We conclude, that antigens of similar molecular-weights can be identified by anti-GBM positive sera in human glomerular, alveolar and placenta basement membranes.
采用凝胶过滤柱对人肾小球基底膜（GBM）、肺泡基底膜（ABM）和胎盘基底膜（PBM）的胶原酶消化产物进行分离，并用抗体抑制酶联免疫吸附测定法（ELISA）鉴定富含Goodpasture抗原（GP）的组分。对每种基底膜制剂进行十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE），随后在硝酸纤维素膜上进行免疫印迹。将活动性GP综合征患者的血清和抗肾小球基底膜抗体（抗GBM抗体）与GBM、ABM和PBM的硝酸纤维素条带孵育。免疫过氧化物酶染色显示，GBM中与24、26、44和50 kD的靶抗原发生反应，ABM和PBM中分别与44和50 kD的靶抗原发生反应。使用恢复期GP血清、其他免疫性疾病患者的血清或健康献血者的血清未观察到相应的反应。这些抗原对还原敏感。我们得出结论，抗GBM阳性血清可在人肾小球、肺泡和胎盘基底膜中鉴定出分子量相似的抗原。
