Graindorge P P, Mahieu P R
Kidney Int. 1978 Dec;14(6):594-606. doi: 10.1038/ki.1978.168.
A radioimmunoassay for detection of antitubular basement membrane (TBM) antibodies was set up using a human TBM antigen (mol wt, 70,000 daltons), purified after collagenase treatment of the insoluble membrane by preparative polyacrylamide electrophoresis, and labeled with iodine 125. Free labeled antigens were separated from those bound to immunoglobulins by a 20% polyethylene glycol (mol wt, 6,000 daltons) solution. In the presence of normal human or Brown Norway rat sera, less than 10% of the labeled antigens were precipitated. In the presence of sera or of kidney eluates from rats immunized with human TBM, the precipitation of the labeled TBM antigens reached 73%, but in the presence of sera from two patients presenting with an interstitial nephritis and linear deposits along the TBM only, up to 47% of the same antigens were precipitated. In these two cases, the anti-TBM antibodies were mainly directed against the heteropolysaccharide-containing glycopeptides isolated from TBM, that is, against the noncollagenous polypeptides of the TBM antigens. Anti-TBM antibodies were sought in the sera of 52 normal blood donors and of 11 patients presenting with glomerulonephritis and linear deposits of immunoglobulins. The average percentage (+/- 1 SD) of labeled TBM antigens precipitated in the serum of normal blood donors was 7.1 +/- 1.2. Of the patients presenting with glomerulonephritis and linear deposits along the GBM, 9 out of 11 exhibited anti-TBM antibodies by radioimmunoassay; among these 9 patients, 8 also displayed linear deposits of IgG along the TBM. Absorption of anti-TBM and anti-GMB antibodies with particulate TBM or GBM, with both types of glycopeptides isolated from GBM or TBM, indicated that the anti-TBM antibodies were directed against the noncollagenous polypeptides of TBM but that the anti-GBM antibodies mainly reacted with the collagenous polypeptides of TBM and GBM. Finally, it was found that the sera of 2 patients out of 15 presenting with lupus nephritis contained a significant anti-TBM-binding activity, mainly directed against the noncollagenous material of TBM.
建立了一种用于检测抗肾小管基底膜（TBM）抗体的放射免疫测定法，该方法使用人TBM抗原（分子量70,000道尔顿），该抗原经胶原酶处理不溶性膜后，通过制备性聚丙烯酰胺电泳纯化，并用碘125标记。通过20%聚乙二醇（分子量6,000道尔顿）溶液将游离标记抗原与结合到免疫球蛋白上的抗原分离。在正常人或棕色挪威大鼠血清存在下，不到10%的标记抗原沉淀。在用人类TBM免疫的大鼠的血清或肾脏洗脱液存在下，标记的TBM抗原沉淀率达到73%，但在仅两名表现为间质性肾炎且沿TBM有线性沉积物的患者的血清存在下，高达47%的相同抗原沉淀。在这两个病例中，抗TBM抗体主要针对从TBM分离的含杂多糖的糖肽，即针对TBM抗原的非胶原多肽。在52名正常献血者和11名患有肾小球肾炎且有免疫球蛋白线性沉积物的患者的血清中寻找抗TBM抗体。正常献血者血清中沉淀的标记TBM抗原的平均百分比（±1标准差）为7.1±1.2。在患有肾小球肾炎且沿肾小球基底膜（GBM）有线性沉积物的患者中，11名中有9名通过放射免疫测定显示有抗TBM抗体；在这9名患者中，8名在TBM上也显示有IgG线性沉积物。用颗粒状TBM或GBM、从GBM或TBM分离的两种糖肽吸收抗TBM和抗GBM抗体表明，抗TBM抗体针对TBM的非胶原多肽，但抗GBM抗体主要与TBM和GBM的胶原多肽反应。最后，发现在15名狼疮性肾炎患者中有2名患者的血清含有显著的抗TBM结合活性，主要针对TBM的非胶原物质。
