Moll R, von Bassewitz D B, Schulz U, Franke W W
Differentiation. 1982;22(1):25-40. doi: 10.1111/j.1432-0436.1982.tb01220.x.
Epithelia-derived tumors (carcinomas) can be distinguished from mesenchymally derived tumors by the presence of intermediate-sized filaments of the cytokeratin type, which usually coincides with the absence of other types of intermediate-sized filaments such as vimentin filaments. In the course of diagnostic examinations of human tumors, using immunofluorescence microscopy, we have come across a case of an unusual carcinoma (Primary tumor and lymph node metastasis) positively stained not only with cytokeratin antibodies but also with immunoglobulins present in vimentin antisera. Therefore, this tumor, a cloacogenic carcinoma apparently derived from the rectal-anal transitional region, has been examined in greater detail using both immunofluorescence microscopy and immuno-electron microscopy as well as gel electrophoretic analysis of cytoskeletal polypeptides from total tumor tissue and from microdissected nodules enriched in carcinoma cells. The unusual reaction of the carcinoma cells with immunoglobulins present in seven different (rabbit or guinea pig) antisera raised against vimentin, has been found to be diminished after absorption on purified cytokeratin or total epidermal cytoskeletal material, but not after absorption on purified vimentin. Gel electrophoretic analysis of tumor cytoskeletons showed an unusual complex pattern of cytokeratin polypeptides containing relatively large (Mr 68,000 and Mr 58,000) neutral-to-slightly basic cytokeratins, as are typically found in epidermis and other stratified squamous epithelia, as well as several smaller acidic cytokeratins, including a Mr 40,000 polypeptide found in certain nonstratified epithelial such as colon and small intestine. Total tumor also showed the inclusion of some vimentin which, however, was significantly decreased in analysis of excised carcinoma nodules. Examining antibody binding to polypeptides separated by gel electrophoresis and blotted on nitrocellulose paper, we have found that antisera raised against vimentin contained not only vimentin antibodies but also immunoglobulins which specifically bound to the largest cytokeratin component. We conclude that the unusual reaction of immunoglobulins present in vimentin antisera with cytokeratin filament bundles does not represent specific binding to vimentin in these carcinoma cells, but is due to a component obviously widespread in vimentin antisera which binds specifically to a cytokeratin present in this type of tumor but not in most other carcinomas. It is proposed that use is made in diagnostic examinations of vimentin antisera or affinity-purified vimentin antibodies that have been pre-absorbed on cytokeratin protein, in order to eliminate such disturbing reactions.
上皮来源的肿瘤（癌）可通过细胞角蛋白类型的中等大小细丝的存在与间充质来源的肿瘤区分开来，这通常与其他类型的中等大小细丝（如波形蛋白细丝）的缺失相一致。在对人类肿瘤的诊断检查过程中，我们利用免疫荧光显微镜遇到了一例不寻常的癌（原发性肿瘤和淋巴结转移），它不仅用细胞角蛋白抗体呈阳性染色，还用波形蛋白抗血清中存在的免疫球蛋白呈阳性染色。因此，对于这个显然起源于直肠肛管过渡区域的泄殖腔源性癌，我们使用免疫荧光显微镜、免疫电子显微镜以及对来自整个肿瘤组织和富含癌细胞的显微切割结节的细胞骨架多肽进行凝胶电泳分析，对其进行了更详细的检查。已发现癌细胞与针对波形蛋白产生的七种不同（兔或豚鼠）抗血清中存在的免疫球蛋白的异常反应，在用纯化的细胞角蛋白或全表皮细胞骨架材料吸收后会减弱，但在用纯化的波形蛋白吸收后则不会减弱。对肿瘤细胞骨架的凝胶电泳分析显示，细胞角蛋白多肽呈现出一种不寻常的复杂模式，其中包含相对较大（分子量68,000和58,000）的中性至微碱性细胞角蛋白，这在表皮和其他复层鳞状上皮中常见，以及几种较小的酸性细胞角蛋白，包括在某些非复层上皮（如结肠和小肠）中发现的分子量40,000的多肽。整个肿瘤还显示含有一些波形蛋白，然而，在对切除的癌结节进行分析时，波形蛋白显著减少。通过检查抗体与经凝胶电泳分离并印迹在硝酸纤维素纸上的多肽的结合情况，我们发现针对波形蛋白产生的抗血清不仅含有波形蛋白抗体，还含有能特异性结合最大细胞角蛋白成分的免疫球蛋白。我们得出结论，波形蛋白抗血清中存在的免疫球蛋白与细胞角蛋白细丝束的异常反应并不代表在这些癌细胞中与波形蛋白的特异性结合，而是由于波形蛋白抗血清中明显广泛存在的一种成分，它特异性结合这种类型肿瘤中存在但大多数其他癌中不存在的一种细胞角蛋白。建议在诊断检查中使用预先用细胞角蛋白蛋白吸收过的波形蛋白抗血清或亲和纯化的波形蛋白抗体，以消除这种干扰反应。
