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-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.
Avanzi
G C
GC
Istituto di Medicina Interna, Universita di Torino, Italy.
Brizzi
M F
MF
Giannotti
J
J
Ciarletta
A
A
Yang
Y C
YC
Pegoraro
L
L
Clark
S C
SC
D000818:Animals; D001681:Biological Assay; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D003470:Culture Media; D016207:Cytokines / Q000494:pharmacology*; D004261:DNA Replication / Q000187:drug effects; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000032:analysis / Q000494:pharmacology*; D006801:Humans; D007106:Immune Sera; D007377:Interleukin-3 / Q000032:analysis / Q000494:pharmacology*; D007700:Kinetics; D007947:Leukemia, Megakaryoblastic, Acute; D011994:Recombinant Proteins / Q000494:pharmacology; D013936:Thymidine / Q000378:metabolism
D003470:Culture Media; D016207:Cytokines; D007106:Immune Sera; D007377:Interleukin-3; D011994:Recombinant Proteins; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D013936:Thymidine