M-07e human leukemic factor-dependent cell line provides a rapid and sensitive bioassay for the human cytokines GM-CSF and IL-3
✍ Scribed by Gian Carlo Avanzi; Maria Felice Brizzi; Joann Giannotti; Agnes Ciarletta; Yu-Chung Yang; Luigi Pegoraro; Steven C. Clark
- Book ID
- 102885802
- Publisher
- John Wiley and Sons
- Year
- 1990
- Tongue
- English
- Weight
- 748 KB
- Volume
- 145
- Category
- Article
- ISSN
- 0021-9541
No coin nor oath required. For personal study only.
✦ Synopsis
We have isolated a subline of the M-07 human megakaryoblastic leukemia cell line, designated M-07e, that requires either interleukin-3 (11-3) 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-y. 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 pgiml 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 colonystimulating 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 11-3 and GM-CSF in regulating hematopoiesis both in patients and in natural sources of the cytokines.
Methods
Cytokines and antisera Homogeneous IL-3 expressed in E. coli and GM-CSF expressed in Chinese hamster ovary cells with specific