M‐07e human leukemic factor‐dependent cell line provides a rapid and sensitive bioassay for the human cytokines GM‐CSF and IL‐3

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‐γ. 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.

[1]  Yu-Chung Yang,et al.  Human IL-3 (multi-CSF): Identification by expression cloning of a novel hematopoietic growth factor related to murine IL-3 , 1986, Cell.

[2]  G. Cattoretti,et al.  Selective growth response to IL‐3 of a human leukaemic cell line with megakaryoblastic features , 1988, British journal of haematology.

[3]  J. Groopman,et al.  Effect of recombinant human granulocyte-macrophage colony-stimulating factor on myelopoiesis in the acquired immunodeficiency syndrome. , 1987, The New England journal of medicine.

[4]  J. Moreau,et al.  Leukaemia inhibitory factor is identical to the myeloid growth factor human interleukin for DA cells , 1988, Nature.

[5]  T. Braciale,et al.  Mycoplasma infection of cell cultures: thymidine incorporation of culture supernatants as a screening test. , 1984, Journal of immunology.

[6]  S. Nagata,et al.  A new bioassay for human granulocyte colony-stimulating factor (hG-CSF) using murine myeloblastic NFS-60 cells as targets and estimation of its levels in sera from normal healthy persons and patients with infectious and hematological disorders. , 1989, Experimental hematology.

[7]  Yu-Chung Yang,et al.  Expression cloning of cDNA encoding a novel human hematopoietic growth factor: human homologue of murine T-cell growth factor P40. , 1989, Blood.

[8]  F. Veglia,et al.  Expression and modulation of IL‐3 and GM‐CSF receptors in human growth factor dependent leukaemic cells , 1990, British journal of haematology.

[9]  A. Ganser,et al.  Recombinant human granulocyte-macrophage colony-stimulating factor in patients with myelodysplastic syndromes--a phase I/II trial. , 1989, Blood.

[10]  D. Caracciolo,et al.  Synergistic and antagonistic effects of recombinant human interleukin (IL) 3, IL-1 alpha, granulocyte and macrophage colony-stimulating factors (G-CSF and M-CSF) on the growth of GM-CSF-dependent leukemic cell lines. , 1987, Journal of immunology.

[11]  K. Miyazono,et al.  Establishment and characterization of a unique human cell line that proliferates dependently on GM‐CSF, IL‐3, or erythropoietin , 1989, Journal of cellular physiology.

[12]  D. Metcalf The hemopoietic colony stimulating factors , 1984 .

[13]  J. Ihle,et al.  Correlation of cell-surface phenotype with the establishment of interleukin 3-dependent cell lines from wild-mouse murine leukemia virus-induced neoplasms. , 1985, Proceedings of the National Academy of Sciences of the United States of America.

[14]  E. Brown,et al.  Human GM-CSF: molecular cloning of the complementary DNA and purification of the natural and recombinant proteins. , 1985, Science.

[15]  J. Gutterman,et al.  Stimulation of myelopoiesis in patients with aplastic anemia by recombinant human granulocyte-macrophage colony-stimulating factor. , 1988, The New England journal of medicine.

[16]  E. Scolnick,et al.  Growth of factor-dependent hemopoietic precursor cell lines , 1980, The Journal of experimental medicine.

[17]  A. Burgess,et al.  Pharmacokinetics of human granulocyte-macrophage colony-stimulating factor using a sensitive immunoassay. , 1988, Blood.

[18]  T. Chen,et al.  In situ detection of mycoplasma contamination in cell cultures by fluorescent Hoechst 33258 stain. , 1977, Experimental cell research.