Biondi A, Roach J A, Griffin J D, Todd R F
Blood. 1985 Jul;66(1):135-42.
To examine the plasma membrane characteristics of an immature monocytic cell capable of proliferation, we have developed a murine monoclonal antibody that identifies an antigen, Mb1, found on the surface of U-937. In immunofluorescence analyses, Mb1 is not expressed by peripheral blood monocytes (freshly isolated, lymphokine-activated, or cultured for seven days), neutrophils, or any other circulating element. It is also absent on human bone marrow mononuclear cells, including the CFU-GM. Among a series of malignant cells from 50 patients with acute myeloid leukemia (including 22 with monocytic or myelomonocytic leukemia), no Mb1 expression was detected. Continuous human cell lines of B or T cell origin were also negative, as were the myeloid lines HL-60 and K562. Apart from U-937, which uniformly expresses Mb1 in high antigen density, only KG-1 (a myeloblastic line) exhibits Mb1 in low antigen density. Exposure of U-937 to phorbol diester (TPA) under conditions that induce features of macrophage differentiation (including the expression of Mo1) results in a significant reduction in Mb1 expression. Mb1 expression is also reduced as a result of culture of U-937 in medium containing anti-Mb1 antibody (antigenic modulation). On sodium dodecyl sulfate-polyacrylamide gel electrophoresis of radiolabeled immunoprecipitates, Mb1 appears to be a dimeric protein with an estimated molecular weight of 80 kd (43 kd under reducing conditions). Antigenic activity on U-937 is destroyed by treatment with trypsin or papain but is regenerated after 24 hours' culture in enzyme-free medium. Mb1 is a constituent plasma membrane protein of U-937, and its degree of expression relates to the state of cellular differentiation.
为了研究能够增殖的未成熟单核细胞的质膜特性,我们制备了一种鼠单克隆抗体,该抗体可识别在U - 937表面发现的一种抗原Mb1。在免疫荧光分析中,外周血单核细胞(新鲜分离的、经淋巴因子激活的或培养7天的)、中性粒细胞或任何其他循环成分均不表达Mb1。人骨髓单个核细胞,包括CFU - GM也不表达Mb1。在50例急性髓系白血病患者(包括22例单核细胞或粒单核细胞白血病患者)的一系列恶性细胞中,未检测到Mb1表达。B或T细胞来源的连续人细胞系也呈阴性,髓系细胞系HL - 60和K562也是如此。除了U - 937以高抗原密度均匀表达Mb1外,只有KG - 1(一种髓母细胞系)以低抗原密度表达Mb1。在诱导巨噬细胞分化特征(包括Mo1表达)的条件下,将U - 937暴露于佛波酯(TPA)会导致Mb1表达显著降低。由于在含有抗Mb1抗体的培养基中培养U - 937(抗原调制),Mb1表达也会降低。在放射性标记免疫沉淀物的十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳中,Mb1似乎是一种二聚体蛋白,估计分子量为80 kd(在还原条件下为43 kd)。用胰蛋白酶或木瓜蛋白酶处理会破坏U - 937上的抗原活性,但在无酶培养基中培养24小时后会再生。Mb1是U - 937的一种组成性质膜蛋白,其表达程度与细胞分化状态有关。