Taha M, Ordoñez N G, Kulkarni S, Owen M, Ro J S, Hortobagyi G, Reading C L, Dicke K A, Spitzer G
Department of Hematology, University of Texas M. D. Anderson Cancer Center, Houston 77030.
Bone Marrow Transplant. 1989 May;4(3):297-303.
We investigated whether monoclonal antibodies (MoAbs) reactive against both acidic and basic cytokeratins alone were sufficient to detect minimal numbers of contaminating epithelial tumor cells in the bone marrow of breast cancer patients. Monoclonal anti-cytokeratin antibodies (AE1 and AE3) were used to stain 14 breast carcinomas by the avidin-biotin-peroxidase technique. Nine tumors (64.3%) showed high reactivity and five (35.7%) showed low or moderate reactivity. Nine MoAbs that proved to be unreactive to light density bone marrow cells by immunoalkaline phosphatase histochemistry were screened for reactivity to breast carcinomas having only low or moderate positivity to cytokeratin antibodies. Three of nine MoAbs showed high percentages of positivity and were selected to supplement the anti-cytokeratin antibodies for immunohistochemical detection of minimal marrow disease in breast cancer patients. A MoAb cocktail was prepared, further tested for reactivity to another five breast carcinomas, and compared with cytokeratin staining alone. The cocktail labeled 100% of carcinoma cells in all the examined specimens. To determine the sensitivity of this panel for detecting minimal numbers of contaminating tumor cells in bone marrow, in vitro mixing experiments were performed. T47D breast carcinoma cells were mixed with bone marrow mononuclear cells at ratios from one tumor cell per 10 bone marrow cells up to one tumor cell per 1 x 10(6) marrow cells, and cytospin preparations were subsequently stained with the MoAb cocktail by the immunoalkaline phosphatase method. Our approach could detect one tumor cell in 1 x 10(5) hematopoietic cells.
我们研究了仅对酸性和碱性细胞角蛋白有反应的单克隆抗体（MoAb）是否足以检测乳腺癌患者骨髓中少量的污染上皮肿瘤细胞。采用抗生物素蛋白-生物素-过氧化物酶技术，用单克隆抗细胞角蛋白抗体（AE1和AE3）对14例乳腺癌进行染色。9例肿瘤（64.3%）显示高反应性，5例（35.7%）显示低或中等反应性。通过免疫碱性磷酸酶组织化学方法筛选出9种对低密度骨髓细胞无反应的单克隆抗体，检测其对细胞角蛋白抗体仅呈低或中等阳性的乳腺癌的反应性。9种单克隆抗体中有3种显示出高阳性率，被选用来补充抗细胞角蛋白抗体，用于免疫组织化学检测乳腺癌患者的微小骨髓疾病。制备了一种单克隆抗体混合物，进一步检测其对另外5例乳腺癌的反应性，并与单独的细胞角蛋白染色进行比较。该混合物在所有检测标本中标记了100%的癌细胞。为了确定该检测方法对检测骨髓中少量污染肿瘤细胞的敏感性，进行了体外混合实验。将T47D乳腺癌细胞与骨髓单个核细胞按每10个骨髓细胞含1个肿瘤细胞至每1×10⁶个骨髓细胞含1个肿瘤细胞的比例混合，随后用免疫碱性磷酸酶方法对细胞涂片制备物进行单克隆抗体混合物染色。我们的方法能够在1×10⁵个造血细胞中检测出1个肿瘤细胞。
