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

Abstract

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.

Recurrent growth factor starvation promotes drug resistance in human leukaemic cells

“…First, MO7e were seeded in a 96-multiwell plate with aliquot volume of 100 ml at the density of 1610 5 ml 71 in the absence of GM-CSF and then cultured at 378C for 2 weeks. Compatible to the previous report that the growth of MO7e completely depends on GM-CSF (Avanzi et al, 1990), the majority of MO7e died within 4 days after GM-CSF deprivation and we could hardly detect the viable cells by direct microscopic observation at day 14. Then we added GM-CSF to each well to expand the viable cells if they existed.…”

“…In the present study, we studied the effects of a transient growth factor deficiency on drug sensitivities using Granulocyte-macrophage colony-stimulating factor (GM-CSF)-dependent human leukaemic MO7e line, which was established from magakaryocytic leukaemia patient as the cells whose growth completely depends on GM-CSF or interleukin-3 (Avanzi et al, 1990). We show that a recurrent growth factor starvation results in a permanent reduction in drug sensitivity.…”

IL-3 is a novel target to interfere with tumor vasculature

“…Informed consent was provided according to the Declaration of Helsinki. M07e cells (Avanzi et al, 1990) and IL-3 releasing gibbon MLA cell line (Brizzi et al, 2001) were used as positive control for c-Kit or IL-3 expression, respectively. Primary renal carcinoma cells were obtained from an informed patient (cells derived from a man 50 years old: T3bG2N0M0) who underwent clinical surgery.…”

A novel approach to identify antigens recognized by CD4 T cells using complement-opsonized bacteria expressing a cDNA library

“…The stimulation of the anti-MAGE-1 DR15-restricted CD4 clone was estimated by testing the capacity of the coculture supernatant to stimulate the proliferation of M-07e cells, which grow in the presence of any of several growth factors, including GM-CSF, IL-3, IL-6 and IL-15. 38,39 Briefly, the clone (3000 cells/ well) was incubated in microwells (100 ml) with EBV-LCL stimulator cells (3 Â 10 4 cells/well) in medium containing IL-2 (25 U/ml). After 24 h, 50 ml of medium were collected and added to M-07e cells (10 4 cells/microwell).…”