Growth regulation of the AML‐193 leukemic cell line: Evidence for autocrine production of granulocyte‐macrophage colony‐stimulating factor (GM‐CSF), and inhibition of GM‐CSF‐dependent cell proliferation by interleukin‐1 (IL‐1) and tumor necrosis factor (tnfα)

Abstract

The human leukemic cell line AML-193 was tested for its proliferative response to endogenously produced autocrine factors and to a variety of cytokines and colony-stimulating factors. Cells grown in the absence of GM-CSF incorporated tritiated thymidine, and this was partially reversed by adding neutralizing anti-GM-CSF antibodies to the culture medium, suggesting that it was due, at least in part, to autocrine GM-CSF production. This was confirmed by immunopurification of a GM-CSF-like activity from cell supernatant of AML-193 cells grown in serum free medium in the absence of exogenous GM-CSF. When AML-193 cells were cultured with GM-CSF in combination with other cytokines, Interleukin-1 alpha and beta (IL-1 alpha and beta), Interleukin-3 (IL-3), Interleukin-6 (IL-6), granulocyte colony-stimulating factor (G-CSF) and tumor necrosis factor alpha (TNF alpha), none of them affected the concentration of GM-CSF required to induce 50% of maximum proliferation (D50). However, the maximum proliferation induced by GM-CSF alone was drastically decreased by IL-1 alpha, IL-1 beta and TNF alpha. Inhibition caused by exposure of the AML-193 to IL-1 for up to 24 hr was reversible, ruling out a direct cytotoxic effect.

Granulocyte‐macrophage colony‐stimulating factor stimulates the metastatic properties of lewis lung carcinoma cells through a protein kinase a signal‐transduction pathway

“…To examine the possibility that rGM-CSF might stimulate the in vitro metastatic properties of LLC-LN7 cells through the PKA second messenger pathway, metastatic LLC-LN7 cells were stably transfected with a mutated PKA R1, gene. Expression of the mutant RI, results in a CAMP-resistant PKA tetramer and, as previously reported, unresponsiveness by the (McKnight et al, 1988;Young et al, 199%). As shown in Figures 5, 7, the REV-LN7 transfectants were less adherent to lung tissue and less invasive through matrigel-coated filters than were the wild-type LLC-LN7 cells.…”

“…Expression of the mutant RI, results in a CAMP-resistant PKA tetramer and, as previously reported, unresponsiveness by the REV-LN7 cells to cAMP elevators (McKnight et al, 1988;Young et al, 199%). As shown in Figures 5, 7, the REV-LN7 transfectants were less adherent to lung tissue and less invasive through matrigel-coated filters than were the wild-type LLC-LN7 cells.…”

“…McKnight, Univ. Washington, Seattle, WA) (McKnight et al, 1988). Also included in the vector was the neomycin phosphotransferase gene which allowed selection of transfected cells by culture in medium containing 400 kg/ml G418.…”

Molecular cloning of a soluble form of the granulocyte‐‐macrophage colony‐stimulating factor receptor α chain from a myelomonocytic cell line. expression, biologic activity, and preliminary analysis of transcript distribution

“…Cell lines and proliferation assay. AML193 cells (23) were obtained from the American Type Culture Collection (ATCC 9589-CRL; Rockville, MD), and M07E cells (24) were from R. Zollner, Genetics Institute (Cambridge, MA). The AML193 cell line was grown serum free in Iscove's modified Dulbecco's medium with insulin (10 pg/ml), transfemn (5-10 unitdml), 1% OPI (oxalate, pyruvate, and insulin) additive, and GM-CSF (0.5 ng/ml).…”

c-KIT expression enhances the leukemogenic potential of 32D cells.

“…In one case the leukaemic cells may produce factors that stimulate the neighboring cells to make growth factors to which the leukemic cells can then respond. For example some human leukaemic cells have been found to produce IL-I which may then induce stromal cells to produce high levels of GM-CSF, G-CSF, and M-CSF (48); these are all factors required for the growth of some leukaemic cells (7,9,(49)(50)(51)(52)(53)(54)(55)(56). This model requires that the leukaemic cells aberrantly express a cytokine and express a receptor for the induced growth factor.…”