Avanzi G C, Brizzi M F, Giannotti J, Ciarletta A, Yang Y C, Pegoraro L, Clark S C
Istituto di Medicina Interna, Universita di Torino, Italy.
J Cell Physiol. 1990 Dec;145(3):458-64. doi: 10.1002/jcp.1041450310.
We have isolated a subline of the M-07 human megakaryoblastic leukemia cell line, designated M-07e, that requires either interleukin-3 (IL-3) or granulocyte macrophage colony-stimulating factor (GM-CSF) for growth, even in the presence of fetal calf serum. This cell line will not grow long term in any other cytokine although it responds slightly to IL-2, IL-4, IL-6, IL-9, and interferon-gamma. We have used the M-07e subline to develop a quantitative bioassay for the measurement of levels of either GM-CSF or IL-3. This assay is as sensitive to either factor as the human bone marrow colony assay (CFU-GM) or the chronic myelogeneous leukemic (CML) blast cell proliferation assay for these factors and is much more convenient and reliable than either. With this assay, as little as 25-50 pg/ml of either IL-3 or GM-CSF can be detected, a level that should render the assay useful for analysis of these molecules in samples from patients undergoing colony-stimulating factor therapy and from conditioned media from natural sources of the factors. In these cases, neutralizing antisera to each cytokine are required to demonstrate the specificity of the assay. This assay, in combination with quantitative immunoassays, should greatly facilitate the analysis of the roles of IL-3 and GM-CSF in regulating hematopoiesis both in patients and in natural sources of the cytokines.
我们分离出了M-07人巨核母细胞白血病细胞系的一个亚系,命名为M-07e,该亚系即使在存在胎牛血清的情况下,生长也需要白细胞介素-3(IL-3)或粒细胞巨噬细胞集落刺激因子(GM-CSF)。尽管它对IL-2、IL-4、IL-6、IL-9和干扰素-γ有轻微反应,但在任何其他细胞因子存在的情况下,该细胞系都不能长期生长。我们利用M-07e亚系开发了一种定量生物测定法,用于测量GM-CSF或IL-3的水平。该测定法对这两种因子的敏感性与人类骨髓集落测定法(CFU-GM)或针对这些因子的慢性粒细胞白血病(CML)原始细胞增殖测定法相同,并且比这两种方法都更方便、更可靠。通过这种测定法,可检测到低至25-50 pg/ml的IL-3或GM-CSF,这一水平应使该测定法可用于分析接受集落刺激因子治疗的患者样本以及这些因子天然来源的条件培养基中的这些分子。在这些情况下,需要针对每种细胞因子的中和抗血清来证明测定法的特异性。这种测定法与定量免疫测定法相结合,应极大地促进对IL-3和GM-CSF在调节患者和细胞因子天然来源的造血过程中的作用的分析。