Miller Y E, Walker S R, Spencer J S, Kubo R T, Mason R J
Department of Medicine, Denver Veterans Administration Medical Center, CO 80220.
Exp Lung Res. 1989 Jul;15(4):635-49. doi: 10.3109/01902148909069623.
Markers specific for various lung cells are useful for studies of cellular differentiation and function. We have produced monoclonal antibodies that bind to isolated rat type II alveolar epithelial cells in an ELISA. Two such antibodies, 2C1 and 3F9, specifically labeled type II cells and nonciliated bronchiolar cells by indirect immunofluorescence of rat lung. A third antibody, 2A3, recognized isolated type II cells by ELISA and immunofluorescence, but did not bind to sections of whole lung. Further immunofluorescence studies on adult rat tissue showed that neither 2C1 nor 3F9 labeled other lung cells or cells in kidney, small intestine, brain, or trachea. The antigen or antigens recognized by 2C1 and 3F9 was not detectable at day 15 of fetal lung gestation but was detectable by day 21. Immunofluorescence studies carried out on 0.5-microns frozen sections of lung tissue demonstrated that both 2C1 and 3F9 bound to cell surface antigens, which are expressed in a highly polarized fashion on the luminal surface of the alveolus and bronchiole. The rat cell line, L2, which displays some similarities to type II cells, did not display positive immunofluorescence to 2A3, 2C1, or 3F9. The antibodies 2C1 and 3F9 are distinct from and apparently more specific than previously described monoclonal antibodies raised to rat type II cells. Alveolar type II and nonciliated bronchiolar cells share several common features. Both cell types contain the surfactant apoprotein SP-A, proliferate in response to lung injury, develop in the late stages of gestation, take up and catabolize platelet-activating factor, contain high levels of cytochrome P-450, and can be induced to form tumors in response to chemical carcinogens. The recognition of highly specific surface antigen(s) on both nonciliated bronchiolar cells and type II cells demonstrates yet another characteristic shared by the two cell types.
各种肺细胞特异性标志物对于细胞分化和功能研究很有用。我们制备了在酶联免疫吸附测定(ELISA)中能与分离的大鼠II型肺泡上皮细胞结合的单克隆抗体。两种这样的抗体,2C1和3F9,通过大鼠肺的间接免疫荧光特异性标记II型细胞和无纤毛细支气管细胞。第三种抗体2A3,通过ELISA和免疫荧光识别分离的II型细胞,但不与全肺切片结合。对成年大鼠组织的进一步免疫荧光研究表明,2C1和3F9都未标记其他肺细胞或肾、小肠、脑或气管中的细胞。2C1和3F9识别的一种或多种抗原在胎肺妊娠第15天时无法检测到,但在第21天时可检测到。对肺组织0.5微米冰冻切片进行的免疫荧光研究表明,2C1和3F9都与细胞表面抗原结合,这些抗原以高度极化的方式在肺泡和细支气管的腔表面表达。大鼠细胞系L2与II型细胞有一些相似之处,但对2A3、2C1或3F9未显示阳性免疫荧光。抗体2C1和3F9与先前针对大鼠II型细胞产生的单克隆抗体不同,且显然更具特异性。肺泡II型细胞和无纤毛细支气管细胞有几个共同特征。这两种细胞类型都含有表面活性物质载脂蛋白SP-A,对肺损伤有增殖反应,在妊娠后期发育,摄取并分解代谢血小板活化因子,含有高水平的细胞色素P-450,并且可被化学致癌物诱导形成肿瘤。在无纤毛细支气管细胞和II型细胞上识别高度特异性的表面抗原证明了这两种细胞类型共有的另一个特征。