Oberneder R, Riesenberg R, Kriegmair M, Bitzer U, Klammert R, Schneede P, Hofstetter A, Riethmüller G, Pantel K
Urologische Universitätsklinik, Klinikum Grosshadern, München, Germany.
Urol Res. 1994;22(1):3-8. doi: 10.1007/BF00431541.
Monoclonal antibodies (mAbs) specific for cytokeratins are potent probes for the identification of disseminated individual epithelial tumour cells in mesenchymal organs such as bone marrow. We have used a monoclonal antibody (mAB) against cytokeratin 18 (CK18) for the detection of individual metastatic tumour cells in bone marrow aspirates from 84 patients with carcinoma of the prostate. CK18+ cells were detected in a sensitivity of 1 per 8 x 10(5) marrow cells using the alkaline phosphatase anti-alkaline phosphatase (APAAP) system for staining. We were able to detect CK18+ tumour cells in the marrow of 33% of patients with stage N0M0 prostate cancers. The incidence of CK18+ cells showed a significant correlation with established risk factors, such as local tumour extent, distant metastases and tumour differentiation. For further characterization of such cells in patients with prostate cancer, we developed an immunocytochemical procedure for simultaneous labelling of cytokeratin component no. 18 (CK18) and prostate-specific antigen (PSA). In a first step, cells were incubated with a murine mAb against PSA, followed by gold-conjugated goat anti-mouse antibodies. In a second step, a biotinylated mAb to CK18 was applied as primary antibody and subsequently incubated with complexes of streptavidin-conjugated alkaline phosphatase, which were developed with Newfuchsin substrate. The binding of gold-labelled antibodies was visualized by silver enhancement. CK18+ cells co-expressing PSA were found in bone marrow aspirates from 5 out of 14 patients with carcinomas of the prostate. The specificity of CK18 for epithelial tumour cells in bone marrow was supported by negative staining of 12 control aspirates from patients with benign prostatic hyperplasia (BPH).(ABSTRACT TRUNCATED AT 250 WORDS)
细胞角蛋白特异性单克隆抗体（mAb）是鉴定骨髓等间充质器官中单个播散上皮肿瘤细胞的有效探针。我们使用抗细胞角蛋白18（CK18）的单克隆抗体（mAB）检测84例前列腺癌患者骨髓穿刺物中的单个转移肿瘤细胞。使用碱性磷酸酶抗碱性磷酸酶（APAAP）染色系统，以每8×10⁵个骨髓细胞中检测到1个的灵敏度检测到CK18⁺细胞。我们能够在33%的N0M0期前列腺癌患者的骨髓中检测到CK18⁺肿瘤细胞。CK18⁺细胞的发生率与已确定的危险因素，如局部肿瘤范围、远处转移和肿瘤分化，呈显著相关性。为了进一步表征前列腺癌患者中的此类细胞，我们开发了一种免疫细胞化学方法，用于同时标记细胞角蛋白成分18（CK18）和前列腺特异性抗原（PSA）。第一步，细胞与抗PSA的鼠源mAb孵育，然后与金标记的山羊抗鼠抗体孵育。第二步，将生物素化的抗CK18 mAb作为一抗应用，随后与链霉亲和素结合碱性磷酸酶复合物孵育，该复合物用新福辛底物显色。通过银增强使金标记抗体的结合可视化。在14例前列腺癌患者的骨髓穿刺物中，有5例发现了共表达PSA的CK18⁺细胞。12例良性前列腺增生（BPH）患者的对照穿刺物染色阴性，支持了CK18对骨髓中上皮肿瘤细胞的特异性。（摘要截断于250字）
