Evans A E, Griffin G C, Tartaglione M, Kennett R H
Hybridoma. 1985 Winter;4(4):289-96. doi: 10.1089/hyb.1985.4.289.
A system is described to detect neuroblastoma (NBL) tumor cells in human bone marrow. The technique exploits the findings that NBL cells have little or no HLA antigen on the surface. Two monoclonal antibodies are used, PI153/3, IgM class, recognizes NBL and some pre-B lymphocytes and KE2 IgG class recognizes HLA. Two second antibodies are used, rhodamine-labeled anti-IgM and fluorescein-labeled anti-IgG. By means of fluorescence microscopy the neuroblastoma cells are labeled with rhodamine only, and the false + pre-B lymphocytes are double labeled with both rhodamine (Rh) and fluorescein (FI) since they are HLA+ and react with KE2. This method has been used to screen the marrow of 24 patients on 40 occasions and 64 laboratory preparations. It is possible to detect NBL cells at a concentration of 1:1000 marrow cells. The advantage of the technique is the fact that false positive cells can be defined because they have HLA surface antigen which neuroblastoma cells do not express.
本文描述了一种用于检测人骨髓中神经母细胞瘤（NBL）肿瘤细胞的系统。该技术利用了NBL细胞表面几乎没有或没有HLA抗原这一发现。使用了两种单克隆抗体，PI153/3，IgM类，识别NBL和一些前B淋巴细胞，KE2 IgG类识别HLA。使用了两种二抗，罗丹明标记的抗IgM和荧光素标记的抗IgG。通过荧光显微镜观察，神经母细胞瘤细胞仅用罗丹明标记，而假阳性的前B淋巴细胞由于它们是HLA阳性并与KE2反应，所以同时用罗丹明（Rh）和荧光素（FI）进行双重标记。该方法已用于对24例患者的骨髓进行40次筛查以及64份实验室标本检测。能够检测到浓度为1:1000骨髓细胞的NBL细胞。该技术的优点是可以定义假阳性细胞，因为它们具有神经母细胞瘤细胞不表达的HLA表面抗原。
