Natori Y, Hayakawa I, Shibata S
Lab Invest. 1986 Jul;55(1):63-70.
Passive Heymann nephritis with acute and severe proteinuria was produced in rats by a single injection of heterologous antibody against a purified glycoprotein which consisted of homologous subunits with a molecular weight of 108,000 (gp108). Gp108 was identified as one of the major antigens in rat renal tubular fraction (FX1A) on immunoblotting assay by using total proteins of FX1A and rabbit antiserum against FX1A. A band of gp330, which was identified as a pathogenic antigen of Heymann nephritis by Kerjaschki D and Farquhar MG (Proc Natl Acad Sci USA 79:5557, 1982) was detected as another band by Coomassie blue staining and immunoblotting. Autoantibodies in the sera of FX1A-injected (active Heymann nephritis) rats reacted to the band of gp330 but not to gp108. These results indicate that gp108 is a different glycoprotein from both gp330 and its degradation products. GP108 was subsequently purified to near homogeneity by extraction with Triton X-100, and then DEAE-cellulose and Bio-Gel A-1.5m column chromatographies. On gel permeation chromatography, the purified antigen showed a molecular weight of 310,000, suggesting that it consists of dimer or trimer of gp108. Rabbits immunized with gp108 produced an antibody which showed monospecific binding to gp108. The antibody stained with brush border of proximal renal tubules in addition to the capillary loops in rat glomeruli by indirect immunofluorescence. Injection of rabbit antiserum against gp108 in rats induced severe proteinuria within 2 days. On the 2nd day after the injection, the glomeruli of the animals showed granular immune deposits along the capillary loops in addition to dominant staining of the brush border of the proximal tubules by immunofluorescence. These results indicate that gp108 is a pathogenic antigen in passive Heymann nephritis and that an antibody against gp108 has a nephritogenic and proteinuria-inducing activity.
通过单次注射针对一种纯化糖蛋白的异源抗体，在大鼠中诱发了伴有急性和严重蛋白尿的被动性海曼肾炎，该糖蛋白由分子量为108,000的同源亚基组成（gp108）。通过使用肾小管部分（FX1A）的总蛋白和抗FX1A兔抗血清进行免疫印迹分析，gp108被鉴定为大鼠肾小管部分（FX1A）中的主要抗原之一。考马斯亮蓝染色和免疫印迹检测到一条gp330条带，Kerjaschki D和Farquhar MG（《美国国家科学院院刊》79:5557，1982）将其鉴定为海曼肾炎的致病抗原。注射FX1A的大鼠（活动性海曼肾炎）血清中的自身抗体与gp330条带反应，但不与gp108反应。这些结果表明，gp108是一种与gp330及其降解产物不同的糖蛋白。随后，通过用Triton X-100提取，然后进行DEAE-纤维素和Bio-Gel A-1.5m柱色谱，将GP108纯化至接近均一。在凝胶渗透色谱上，纯化的抗原显示分子量为3,10,000，表明它由gp108的二聚体或三聚体组成。用gp108免疫的兔子产生了一种抗体，该抗体显示出与gp108的单特异性结合。通过间接免疫荧光，该抗体除了在大鼠肾小球的毛细血管袢上染色外，还在近端肾小管的刷状缘上染色。给大鼠注射抗gp108兔抗血清在2天内诱发了严重蛋白尿。注射后第2天，动物的肾小球除了近端肾小管刷状缘的主要染色外，还在毛细血管袢上显示颗粒状免疫沉积物。这些结果表明，gp108是被动性海曼肾炎中的致病抗原，并且抗gp108抗体具有致肾炎和诱导蛋白尿的活性。
