Sperber K, Gollub E, Goswami S, Kalb T H, Mayer L, Marom Z
Department of Medicine, Mount Sinai Medical Center, New York, New York.
Am Rev Respir Dis. 1992 Dec;146(6):1589-97. doi: 10.1164/ajrccm/146.6.1589.
We previously described a novel 68,000 D macrophage-derived protein (MMS-68) that can stimulate mucus-like glycoconjugate (MLGC) secretion from cultured human airways, respiratory epithelial cells, and the ishikawa adenocarcinoma cell line. To better characterize this mucus secretagogue, we generated monoclonal antibodies against MMS-68 by injecting crushed SDS-PAGE gel slices containing this protein into Balb-C mice followed by fusion with SP2/0, a nonsecreting mouse myeloma cell line. A panel of monoclonal antibodies was produced that identified the 68,000 D MMS by immunoblot analysis and immunoprecipitation. The monoclonal antibodies detected MMS-68 in normal peripheral blood monocytes and pulmonary macrophages by cytofluorographic analysis and in human airways as determined by immunohistochemistry. Utilizing the monoclonal antibodies, an antigen-capture ELISA assay was developed. Statistically significant elevations in levels of MMS-68 were detected in bronchoalveolar lavage fluid (BALF) of chronic bronchitic subjects and cigarette smokers and in monocyte culture supernatants from steroid-dependent asthmatic patients compared to normal control subjects. The 68,000 D MMS is a potent secretagogue and may play an important role in the regulation of mucus secretion, especially in chronic bronchitis and steroid-dependent asthma.
我们之前描述了一种新型的68000 D巨噬细胞衍生蛋白（MMS - 68），它能刺激培养的人呼吸道、呼吸道上皮细胞和石川腺癌细胞系分泌黏液样糖缀合物（MLGC）。为了更好地表征这种黏液分泌刺激物，我们通过将含有该蛋白的破碎SDS - PAGE凝胶切片注射到Balb - C小鼠体内，随后与非分泌型小鼠骨髓瘤细胞系SP2/0融合，产生了针对MMS - 68的单克隆抗体。通过免疫印迹分析和免疫沉淀产生了一组能识别68000 D MMS的单克隆抗体。通过细胞荧光分析，单克隆抗体在正常外周血单核细胞和肺巨噬细胞中检测到MMS - 68，通过免疫组织化学在人呼吸道中也检测到了该蛋白。利用这些单克隆抗体，开发了一种抗原捕获ELISA检测法。与正常对照受试者相比，在慢性支气管炎患者和吸烟者的支气管肺泡灌洗液（BALF）以及类固醇依赖型哮喘患者的单核细胞培养上清液中，检测到MMS - 68水平有统计学意义的升高。68000 D MMS是一种有效的分泌刺激物，可能在黏液分泌的调节中起重要作用，尤其是在慢性支气管炎和类固醇依赖型哮喘中。
