Bühring H J, Müller T, Herbst R, Cole S, Rappold I, Schuller W, Zhu X, Fritzsch U, Faul C, Armeanu S, Ullrich A, Klein G, Schmidt H
Second Department of Internal Medicine, University of Tübingen, Germany.
Leukemia. 1996 Jan;10(1):106-16.
The antigen expression of immature erythroid bone marrow cells was studied using two recently generated monoclonal antibodies (mAb), mAb 67A4 and 9C4, with specificities for the epithelial cell adhesion molecule E-cadherin (E-cad; mAb 67A4), and a novel 110 kDa differentiation antigen (mAb 9C4) with unknown molecular structure. Pappenheim staining of FACS-purified cells labeled with mAb 9C4 and anti-glycophorin A (GA) revealed that the majority of the 9C4+GA- and 9C4+GA+ cells consisted of erythroblasts. In contrast, the E-cad-positive population comprised normoblasts and erythroblasts. While the E-cad+GA- fraction contained mainly erythroblasts and basophilic normoblasts, the E-Cad+GA+ population was enriched in orthochromatic and polychromatophilic normoblasts. By colony assays of affinity column-purified cells it could be shown that erythroid colony forming units (CFU-E) were enriched and erythroid burst forming units (BFU-E) were depleted in the 9C4- and E-cad-positive fractions. Flow cytometric analysis of bone marrow cells double-labeled with mAb 67A4 and anti-CD71, anti-CD117, anti-CD34, or anti-GA revealed that about 90% of the E-cad-positive cells coexpressed CD71, about 70% were positive for CD117, about 50% for GA, and only about 5% coexpressed CD34. The expression pattern of 9C4 antigen was similar to that of E-Cad with the exception that only a minority of the 9C4-positive cells coexpressed GA. Lymphoid and myeloid markers were negative on both the E-Cad- and 9C4-positive populations. In these studies we describe the identification of a new mAb-defined antigen which is specifically expressed on erythroblasts and CFU-E(9C4) and demonstrate that E-Cad is not only expressed on epithelial cells but also on erythropoietic cells of defined maturational stages.
利用两种新产生的单克隆抗体(mAb),即对上皮细胞粘附分子E-钙粘蛋白(E-cad;mAb 67A4)具有特异性的mAb 67A4和对一种分子结构未知的新型110 kDa分化抗原(mAb 9C4),研究了未成熟红系骨髓细胞的抗原表达。用mAb 9C4和抗血型糖蛋白A(GA)标记的FACS纯化细胞的帕彭海姆染色显示,大多数9C4+GA-和9C4+GA+细胞由成红细胞组成。相比之下,E-钙粘蛋白阳性群体包括正成红细胞和成红细胞。虽然E-cad+GA-部分主要包含成红细胞和嗜碱性正成红细胞,但E-Cad+GA+群体富含正染和多染正成红细胞。通过对亲和柱纯化细胞的集落分析表明,在9C4-和E-钙粘蛋白阳性部分中,红系集落形成单位(CFU-E)富集,红系爆式集落形成单位(BFU-E)减少。用mAb 67A4和抗CD71、抗CD117、抗CD34或抗GA对骨髓细胞进行双标记的流式细胞术分析显示,约90%的E-钙粘蛋白阳性细胞共表达CD71,约70%为CD117阳性,约50%为GA阳性,只有约5%共表达CD34。9C4抗原的表达模式与E-钙粘蛋白相似,不同之处在于只有少数9C4阳性细胞共表达GA。E-钙粘蛋白阴性和9C4阳性群体上的淋巴样和髓样标志物均为阴性。在这些研究中,我们描述了一种新的mAb定义抗原的鉴定,该抗原在成红细胞和CFU-E(9C4)上特异性表达,并证明E-钙粘蛋白不仅在上皮细胞上表达,而且在特定成熟阶段的造血细胞上表达。