Azzarone B, Malpièce Y, Zaech P, Moretta L, Fauci A, Suarez H
Exp Cell Res. 1985 Aug;159(2):451-62. doi: 10.1016/s0014-4827(85)80018-5.
4F2 monoclonal antibody recognizes a 120-kD glycoprotein on the surface of human spread fibroblastic cells of embryonic and neoplastic origin, but it does not bind to normal spread adult fibroblasts. Flow cytometric analysis reveals that human adult fibroblasts become 4F2-positive when they are analyzed as round-shaped cells; this means that, in normal adult cells, 4F2 antigen behaves as a cryptic molecule. Thus, the basic difference between embryonic, neoplastic and normal adult cells consists in a different organization in the architecture of the cell membrane, since in embryonic and neoplastic cells there is a continuous expression of the 4F2 antigen independently of the cell shape and cell cycle phase. Quantitative flow cytometry shows that the mean surface density (MSD) of the 4F2 antigen 1, does not vary as a function of the cell cycle; 2, is inversely related to cell size and "metabolic time". This suggests that at the plateau phase the surface organization of G1 resting cells changes as a function of the number of days spent in culture; and 3, sarcoma and SV40-transformed cells show significantly increased MSD levels of the 4F2 antigen in comparison with normal cells of similar size. Electrophoretic analysis under reducing conditions confirms the quantitative differences in the expression of the 4F2 antigen described with the cell sorter. It also reveals, in a way different from that previously found with lymphoid cells, the coexistence of two molecules (85 and 73 kD) in the heavy chain regions. The 73 kD is, however, much more strongly expressed in the fibrosarcoma than in the embryonic cells. Finally, it shows that 4F2 antigen is a very useful tool for studying the organization and the structure of the cell membrane of human fibroblasts and can provide new insights to understand better the developmental and transformation processes.
4F2单克隆抗体可识别源自胚胎和成瘤性的人传代成纤维细胞表面的一种120-kD糖蛋白,但不与正常传代的成人成纤维细胞结合。流式细胞术分析显示,当成人成纤维细胞呈圆形细胞进行分析时会变为4F2阳性;这意味着,在正常成人细胞中,4F2抗原表现为一种隐蔽分子。因此,胚胎细胞、肿瘤细胞和正常成人细胞之间的基本差异在于细胞膜结构组织的不同,因为在胚胎细胞和肿瘤细胞中,4F2抗原的表达是持续的,与细胞形状和细胞周期阶段无关。定量流式细胞术表明,4F2抗原的平均表面密度(MSD):1. 不随细胞周期而变化;2. 与细胞大小和“代谢时间”呈负相关。这表明在平台期,G1期静止细胞的表面组织随培养天数而变化;3. 与大小相似的正常细胞相比,肉瘤细胞和SV40转化细胞的4F2抗原MSD水平显著升高。还原条件下的电泳分析证实了用细胞分选仪描述的4F2抗原表达的定量差异。它还以一种不同于先前在淋巴细胞中发现的方式揭示了重链区域中两种分子(85 kD和73 kD)的共存。然而,73 kD在纤维肉瘤中的表达比在胚胎细胞中强得多。最后,结果表明4F2抗原是研究人成纤维细胞膜的组织和结构的一种非常有用的工具,可为更好地理解发育和转化过程提供新的见解。