Settle S A, Hellström I, Hellström K E
Exp Cell Res. 1985 Apr;157(2):293-306. doi: 10.1016/0014-4827(85)90114-4.
Monoclonal antibodies were generated by immunizing rats with mouse bladder carcinomas, fusing their spleen cells with NS-1 myeloma cells and selecting for antibodies that bound to mouse bladder carcinomas. One of the antibodies, IG5, recognizes an antigenic determinant which is present in mouse bladder carcinoma cytoskeletons and is not detectable in the normal bladder epithelium. Indirect immunofluorescence studies revealed that the antigen is expressed intracellularly and is organized in the form of filamentous arrays. The antigen was detected by peroxidase--antiperoxidase immunocytochemistry in stratified epithelia and glands derived from these, but has not been observed in any tissues of mesenchymal or neuronal origin. Various normal and neoplastic human tissues were subsequently tested for reactivity with antibody IG5. Antigen expression in normal tissues was similar to that in the mouse. Most carcinomas of the bladder and lung were stained, while all of eleven colon carcinomas were negative. Antibody IG5 immunoprecipitated radio-iodinated peptides of 58, 56, 52 and 43 kD molecular weight from mouse bladder carcinomas. Western blotting experiments with antibody IG5 demonstrated bands of 56 and 50 kD in a keratin-enriched fraction of the bladder carcinoma cytoskeleton. Antibody IG5 reacted with molecules which have several properties typical of cytoskeletal keratin peptides. Our findings are discussed in the context of previously described keratin peptides and relevant monoclonal antibodies.
通过用小鼠膀胱癌免疫大鼠、将其脾细胞与NS - 1骨髓瘤细胞融合并筛选与小鼠膀胱癌结合的抗体来产生单克隆抗体。其中一种抗体IG5识别一种抗原决定簇，该抗原决定簇存在于小鼠膀胱癌的细胞骨架中，在正常膀胱上皮中无法检测到。间接免疫荧光研究表明，该抗原在细胞内表达，并以丝状阵列的形式组织。通过过氧化物酶 - 抗过氧化物酶免疫细胞化学在源自这些组织的复层上皮和腺体中检测到该抗原，但在间充质或神经源性的任何组织中均未观察到。随后测试了各种正常和肿瘤性人类组织与抗体IG5的反应性。正常组织中的抗原表达与小鼠中的相似。大多数膀胱癌和肺癌被染色，而11例结肠癌全部为阴性。抗体IG5从小鼠膀胱癌中免疫沉淀出分子量为58、56、52和43 kD的放射性碘化肽。用抗体IG5进行的蛋白质免疫印迹实验在膀胱癌细胞骨架富含角蛋白的部分中显示出56和50 kD的条带。抗体IG5与具有几种细胞骨架角蛋白肽典型特性的分子发生反应。我们的研究结果将在先前描述的角蛋白肽和相关单克隆抗体的背景下进行讨论。
