Makker S P, Singh A K
Lab Invest. 1984 Mar;50(3):287-93.
The nephritogenic brush border glycoprotein antigen (gp600) of autoimmune membranous glomerulonephropathy (Heymann nephritis) isolated from crude proximal renal tubular membrane fraction (FX1A) by Lens culinaris lectin affinity chromatography was characterized. The molecular weight of the antigen determined by molecular exclusion chromatography was found to be 600,000. On sodium dodecyl sulfate-polyacrylamide gel electrophoresis it resolved into five polypeptides which were in the molecular weight range of 70,000 to 330,000. All five polypeptide bands stained for carbohydrates and lipids with Schiff's reagent and Sudan black, respectively. Rabbits immunized with gp600 produced an antibody that specifically reacted with the brush border of proximal renal tubules of rat by indirect immunofluorescence and gave a single precipitin line in double diffusion in gel (Ouchterlony) against the antigen. The specificity of gp600 was proven by (a) the ability of gp600 to produce active Heymann nephritis, (b) the ability of anti-gp600 to produce passive Heymann nephritis, and (c) the ability of gp600 alone, out of other antigens in FX1A, to specifically react with the nephritogenic autoantibody. The latter was tested by an enzyme immunodiffusion technique described in this report.
对通过扁豆凝集素亲和层析从粗制近端肾小管膜组分（FX1A）中分离得到的自身免疫性膜性肾小球肾炎（海曼肾炎）的致肾炎刷状缘糖蛋白抗原（gp600）进行了特性鉴定。通过分子排阻色谱法测定的该抗原分子量为600,000。在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳上，它分解为五条分子量在70,000至330,000范围内的多肽。所有五条多肽带分别用席夫试剂和苏丹黑对碳水化合物和脂质进行了染色。用gp600免疫的兔子产生了一种抗体，该抗体通过间接免疫荧光法与大鼠近端肾小管的刷状缘发生特异性反应，并且在凝胶双向扩散（欧氏）中与该抗原形成单一沉淀线。gp600的特异性通过以下几点得到证明：（a）gp600产生活动性海曼肾炎的能力；（b）抗gp600产生被动性海曼肾炎的能力；（c）在FX1A的其他抗原中，仅gp600能与致肾炎自身抗体发生特异性反应。后者通过本报告中描述的酶免疫扩散技术进行了检测。
