Nakamura T, Oite T, Kazama T, Suzuki S, Orikasa M, Arakawa M, Shimizu F
Department of Immunology, Nigata University School of Medicine, Japan.
Virchows Arch A Pathol Anat Histopathol. 1988;412(6):573-82. doi: 10.1007/BF00844293.
Using cultured human fetal kidney cortical cells as antigen, two monoclonal antibodies (moAbs) against human glomeruli were produced. One of these moAbs, H-4, recognized the cell surface of glomerular epithelial cells, and the other, H-13, recognized the extracellular matrix present in the mesangial area. Both also reacted with liver, H-4 recognizing antigen present on the hepatocyte, and H-13 recognizing antigen distributed along the sinusoid. Species specificity for these moAbs was examined using mouse, rat, guinea pig and rabbit glomeruli, which revealed that H-4 reacted with rat glomerular epithelial cells and H-13 stained guinea pig glomerular mesangium. In the human fetal kidney, H-13 reacted with the mesangium, glomerular and tubular basement membrane and Bowman's capsule, and H-4 with the glomerular and tubular epithelial cells. Dot immunobinding assay of fibronectin purified from glomerular culture supernatant and plasma revealed that H-13 recognized both plasma and cellular fibronectin. Immunoblot analysis of 2.0 M guanidine HCl extract after dissociation in sodium dodecyl sulfate and electrophoresis demonstrated binding of H-4 to a 125 kd polypeptide. Immunoblot analysis of thermolysin-digested fibronectin exhibited binding of H-13 to 145 kd and 110 kd fragments, but not to 38 kd-29 kd fragments. In renal biopsy specimens from patients with membranous nephropathy. H-13 stained the glomerular basement membrane (GBM), but not the mesangium, whereas anti-fibronectin antisera stained both the GBM and the mesangium. In those from patients with minimal change nephrotic syndrome (MCNS), IgA glomerulonephritis (IgAGN) and membranoproliferative glomerulonephritis (MPGN), the staining pattern with H-13 was similar to that with polyclonal anti-fibronectin antisera. These results indicate that H-4 recognizes a 125 kd polypeptide constituent of the glomerular epithelial cell membrane and that H-13 recognizes the cell binding domain of fibronectin as well as revealing structural alterations in the mesangium and GBM.
以培养的人胎儿肾皮质细胞作为抗原，制备了两种抗人肾小球的单克隆抗体（moAbs）。其中一种单克隆抗体H - 4识别肾小球上皮细胞的细胞表面，另一种H - 13识别系膜区存在的细胞外基质。二者也与肝脏发生反应，H - 4识别肝细胞上存在的抗原，H - 13识别沿肝血窦分布的抗原。使用小鼠、大鼠、豚鼠和兔的肾小球检测这些单克隆抗体的种属特异性，结果显示H - 4与大鼠肾小球上皮细胞发生反应，H - 13使豚鼠肾小球系膜染色。在人胎儿肾中，H - 13与系膜、肾小球和肾小管基底膜以及鲍曼囊发生反应，H - 4与肾小球和肾小管上皮细胞发生反应。对从肾小球培养上清液和血浆中纯化的纤连蛋白进行斑点免疫结合试验表明，H - 13识别血浆和细胞纤连蛋白。对在十二烷基硫酸钠中解离并电泳后的2.0M盐酸胍提取物进行免疫印迹分析表明，H - 4与一条125kd的多肽结合。对嗜热菌蛋白酶消化的纤连蛋白进行免疫印迹分析显示，H - 13与145kd和110kd的片段结合，但不与38kd - 29kd的片段结合。在膜性肾病患者的肾活检标本中，H - 13使肾小球基底膜（GBM）染色，但不使系膜染色，而抗纤连蛋白抗血清使GBM和系膜均染色。在微小病变肾病（MCNS）、IgA肾小球肾炎（IgAGN）和膜增生性肾小球肾炎（MPGN）患者的标本中，H - 13的染色模式与多克隆抗纤连蛋白抗血清的染色模式相似。这些结果表明，H - 4识别肾小球上皮细胞膜的一种125kd多肽成分，H - 13识别纤连蛋白的细胞结合结构域，同时也揭示了系膜和GBM的结构改变。
