Nüsing R, Ullrich V
Faculty of Biology, University of Konstanz, Federal Republic of Germany.
Eicosanoids. 1990;3(3):175-80.
Although the formation of TxB2 has been reported for different cells such as monocytes, lymphocytes, granulocytes or fibroblasts and tissues like lung and liver, the corresponding enzyme has not yet been measured. We have established an enzyme-linked immunosorbent assay (ELISA) for measurement of human Tx-synthase with a monospecific polyclonal and a purified monoclonal antibody. ELISA data were verified by either western blots or immunohistochemistry. Next to platelets, blood monocytes were found to have the highest content of Tx-synthase, whereas lung fibroblasts and promyelocytic cells possess only very low levels. In granulocytes and lymphocytes neither Tx-synthase nor its activity could be demonstrated. Among different tissues, lung and liver showed the highest content of Tx-synthase. These data correlate TxB2-synthase activity in a given human tissue with its enzyme content of Tx-synthase for the first time. In view of the frequent involvement of TxA2 in pathological conditions, the technique developed may prove valuable for the characterization of disease states, particularly inflammatory events.
尽管已有报道称不同细胞(如单核细胞、淋巴细胞、粒细胞或成纤维细胞)以及肺和肝等组织可生成血栓素B2(TxB2),但尚未对相应的酶进行检测。我们已经建立了一种酶联免疫吸附测定(ELISA)方法,用于使用单特异性多克隆抗体和纯化的单克隆抗体检测人Tx合成酶。ELISA数据通过蛋白质免疫印迹法或免疫组织化学法进行验证。除血小板外,发现血液单核细胞的Tx合成酶含量最高,而肺成纤维细胞和早幼粒细胞的含量极低。在粒细胞和淋巴细胞中,既未检测到Tx合成酶,也未检测到其活性。在不同组织中,肺和肝的Tx合成酶含量最高。这些数据首次将给定人体组织中的TxB2合成酶活性与其Tx合成酶的酶含量相关联。鉴于血栓素A2(TxA2)频繁参与病理状况,所开发的技术可能对疾病状态的表征,特别是炎症事件具有重要价值。