Mew D, Lum V, Wat C K, Towers G H, Sun C H, Walter R J, Wright W, Berns M W, Levy J G
Cancer Res. 1985 Sep;45(9):4380-6.
A variety of cell lines have been prepared by fusion of the murine WEH1 3B cell line with peripheral blood leukocytes from a patient with chronic granulocytic leukemia. Fusion products were selected for their ability to produce a leukemia-associated antigen (CAMAL) previously described. One such line which originally produced CAMAL subsequently lost this ability and was used as a negative control. A number of antibodies were conjugated to hematoporphyrin (HP) and tested for their ability to bind to cell lines as detected by either fluorescence or by their ability to kill cells after light activation. The antibodies used were: rabbit anti-Hu (a conventional rabbit antiserum raised to membrane preparations from normal human peripheral blood leukocytes which served as a positive control); CAMAL-1 (a monoclonal gamma 1 antibody with specificity for the CAMAL antigen); and L1210 (an irrelevant monoclonal gamma 1 antibody). HP was conjugated to the antibodies by a carbodiimide procedure. When labeled cells were examined by fluorescence microscopy, it was apparent that both the rabbit antibody and CAMAL-1:HP showed positive labeling. The ability of the antibody:HP conjugates to kill labeled cells following light activation was tested. It was shown that rabbit anti-Hu:HP and CAMAL-1:HP conjugates were capable of killing significant numbers of cells when HP concentrations were as low as 1.2 ng/10(6) cells, whereas similarly treated cells exposed to either L1210:HP or HP alone did not exhibit significant killing until concentrations reached 240 and 120 ng/10(6) cells, respectively. Further experiments in which other cell lines were tested, all at HP concentrations of 12 ng/10(6) cells, demonstrated that those lines producing CAMAL were killed, whereas negative lines were not.
通过将小鼠WEH1 3B细胞系与一名慢性粒细胞白血病患者的外周血白细胞融合,制备了多种细胞系。融合产物因其产生先前描述的白血病相关抗原(CAMAL)的能力而被筛选出来。其中一个最初产生CAMAL的细胞系后来失去了这种能力,并被用作阴性对照。一些抗体与血卟啉(HP)偶联,并测试它们与细胞系结合的能力,通过荧光检测或光激活后杀死细胞的能力来检测。所使用的抗体如下:兔抗人(一种常规兔抗血清,由正常人外周血白细胞的膜制剂制备而成,用作阳性对照);CAMAL-1(一种对CAMAL抗原有特异性的单克隆γ1抗体);以及L1210(一种无关的单克隆γ1抗体)。通过碳二亚胺法将HP与抗体偶联。当通过荧光显微镜检查标记细胞时,很明显兔抗体和CAMAL-1:HP均显示阳性标记。测试了抗体:HP偶联物在光激活后杀死标记细胞的能力。结果表明,当HP浓度低至1.2 ng/10(6)细胞时,兔抗人:HP和CAMAL-1:HP偶联物能够杀死大量细胞,而暴露于L1210:HP或单独HP的同样处理的细胞,直到浓度分别达到240和120 ng/