Morikawa K, Morikawa S, Miyawaki T, Nagasaki M, Torii I, Imai K
Department of Internal Medicine, Shimane Medical University, Japan.
Br J Haematol. 1996 Aug;94(2):250-7. doi: 10.1046/j.1365-2141.1966.d01-1806.x.
The present study demonstrated that a human B-cell line derived from non-Hodgkin's lymphoma. HCF-MLpN. constitutively expressed G-CSF receptor on the cell surface. G-CSF binding to the cell surface was shown by immunofluorescence staining using biotinylated G-CSF preparation and analysed by flow cytometry. Specific binding of G-CSF to the cells was shown by pretreatment with unlabelled G-CSF. In the radioreceptor assay and Scatchard plot analysis using radiolabelled ligand, MLpN cells revealed a single species of binding site with an equilibrium dissociation constant of 167 (153-182) pM and a maximal binding site per cell of 1076 (1044-1116). The G-CSF receptor mRNA transcript was exhibited in the RNA from MLpN cells by reverse transcriptase polymerase chain reaction procedure. [3H]thymidine incorporation and trypan blue exclusion showed that the G-CSF receptor was capable of transducing the growth signal to HCF-MLpN cells. A small fraction of fresh B blasts from six patients with B-cell lymphoma and leukaemia displayed G-CSF binding by two-colour immunofluorescence staining. In contrast, a panel of seven B-cell lines was negative for the binding to biotinylated G-CSF preparation. These results suggest that the phenotype of G-CSF binding may be lost during the culture. The expression of G-CSF receptor in HCF-MLpN cells appeared to be exceptional.
本研究表明，一种源自非霍奇金淋巴瘤的人B细胞系HCF-MLpN在细胞表面组成性表达G-CSF受体。使用生物素化的G-CSF制剂通过免疫荧光染色显示G-CSF与细胞表面的结合，并通过流式细胞术进行分析。未标记的G-CSF预处理显示了G-CSF与细胞的特异性结合。在使用放射性标记配体的放射受体分析和Scatchard图分析中，MLpN细胞显示出单一类型的结合位点，平衡解离常数为167（153 - 182）pM，每个细胞的最大结合位点为1076（1044 - 1116）。通过逆转录聚合酶链反应程序在MLpN细胞的RNA中检测到G-CSF受体mRNA转录本。[3H]胸苷掺入和台盼蓝排斥试验表明，G-CSF受体能够将生长信号转导至HCF-MLpN细胞。通过双色免疫荧光染色，来自6例B细胞淋巴瘤和白血病患者的一小部分新鲜B淋巴细胞显示出G-CSF结合。相比之下，一组7种B细胞系对生物素化G-CSF制剂的结合呈阴性。这些结果表明，G-CSF结合的表型在培养过程中可能会丧失。HCF-MLpN细胞中G-CSF受体的表达似乎是例外情况。
