Berois N, Varangot M, Osinaga E, Babino A, Caignault L, Musé I, Roseto A
Depto. de Bioquímica, Facultad de Medicina, Montevideo, Uruguay.
Anticancer Res. 1997 Jul-Aug;17(4A):2639-46.
The detection of occult carcinoma cells in patients with breast cancer may aid determination of prognosis and the development of new therapeutic approaches. In this study, we report a new method to detect rare human breast cancer cells, which combines an immunomagnetic separation (IMS) procedure with cytokeratin 19 (CK 19) immunostaining.
Four monoclonal antibodies (MAb) previously characterized against cell surface antigens (1BE12, ED8, 7B10 and 83D4), were evaluated for IMS optimization. Immunoseparated epithelial cells were identified using a MAb against CK 19. We compared the IMS procedure with the immunocytochemistry (ICC) and the RT-PCR for CK 19 on an "in vitro" experimental model.
The best results in IMS procedures were obtained using MAbs 1BE12 (directed against Lewis y antigen) and ED8 (directed against MUC 1). In reconstitution experiments, using several ratios of T47D cells mixed with peripheral-blood mononuclear (PBMN) cells, the IMS procedure reliably detects one mammary carcinoma cell in 5 x 10(5) PBMN cells, whereas the ICC detects up to one T47D cell per 10(5) PBMN cells. The best sensitivity was observed with the RT-PCR (up to one T47D cell per 10(6) PBMN cells). We found the same high specificity with the three methods evaluated.
The IMS procedure using MAbs 1BE12 or ED8 associated with CK 19 immunostaining is a specific, sensitive, and feasible method for the detection of rare human breast cancer cells. This method proved to be better than the ICC staining but its sensitivity was lower than that of RT-PCR for CK 19.
检测乳腺癌患者体内隐匿的癌细胞有助于判断预后及开发新的治疗方法。在本研究中，我们报告了一种检测罕见人类乳腺癌细胞的新方法，该方法将免疫磁珠分离（IMS）程序与细胞角蛋白19（CK 19）免疫染色相结合。
评估了四种先前针对细胞表面抗原（1BE12、ED8、7B10和83D4）鉴定的单克隆抗体（MAb）用于IMS优化。使用抗CK 19的单克隆抗体鉴定免疫分离的上皮细胞。我们在“体外”实验模型上比较了IMS程序与免疫细胞化学（ICC）以及CK 19的逆转录聚合酶链反应（RT-PCR）。
使用单克隆抗体1BE12（针对Lewis y抗原）和ED8（针对MUC 1）在IMS程序中获得了最佳结果。在重组实验中，使用几种比例的T47D细胞与外周血单个核（PBMN）细胞混合，IMS程序能可靠地在5×10⁵个PBMN细胞中检测到一个乳腺癌细胞，而ICC每10⁵个PBMN细胞最多检测到一个T47D细胞。RT-PCR观察到最佳灵敏度（每10⁶个PBMN细胞中最多一个T47D细胞）。我们发现所评估的三种方法具有相同的高特异性。
使用与CK 19免疫染色相关的单克隆抗体1BE12或ED8的IMS程序是检测罕见人类乳腺癌细胞的一种特异、灵敏且可行的方法。该方法被证明优于ICC染色，但其灵敏度低于CK 19的RT-PCR。
