Biosynthetic (recombinant) human granulocyte-macrophage colony- stimulating factor: effect on normal bone marrow and leukemia cell lines

Abstract

To examine the biologic properties of the molecule encoded by the human gene for granulocyte-macrophage colony-stimulating factor (GM-CSF), we expressed the cloned complementary DNA (cDNA) in transfected monkey COS cells and purified the resultant protein. Purified biosynthetic human GM-CSF was added to cultures of normal hematopoietic progenitor cells in semisolid media, and the resulting colonies were characterized cytochemically. Non-adherent light-density bone marrow cells from healthy adult volunteers were maximally stimulated with GM-CSF (approximately 250 pmol/L, and four types of colonies were consistently identified by aspirating the individual colonies and staining with a triple stain for specific and nonspecific esterases and eosinophilic granules. Pure neutrophilic granulocyte (G), mixed granulocyte- macrophage (GM), pure macrophage (M), and pure eosinophil (EO) colonies were observed, the mean incidences on day 8 being 70%, 20%, 5%, and 5%, and on day 14, 7.5%, 16.6%, 50.9%, and 25.0%, respectively. In all types of colonies, complete maturation to segmented forms or typical macrophages was detected. GM-CSF did not enhance the growth of BFU-E from normal peripheral blood buffy coat cells in the simultaneous presence of erythropoietin alone or erythropoietin with purified erythroid-potentiating activity. GM-CSF stimulated HL-60 and KG-1 colony formation twofold and fivefold, respectively; consistent differentiation induction towards monocytic and eosinophilic lineages was observed in HL-60 but not in KG-1. These in vitro findings indicate that GM-CSF is a multilineage stimulator for progenitor cells of G, GM, M, and EO colonies.

Effect of recombinant human interleukin 3, granulocyte‐macrophage colony‐stimulating factor and granulocyte colony‐stimulating factor on human bfu‐e in serum‐free cultures

“…We have shown that rhGM-CSF alone did not induce human burst formation and could not augment the effect of rhEpo on human BFU-e growth. In this regard, earlier reports on murine [25] and human [24, cultures are controversial in that only some investigators are in agreement with our results [25,26]. On the other hand, other investigators have reported that GM-CSF stimulates BFU-e in the presence of Epo, or has BPA [24,[27][28][29][30][31].…”

Induction of colony‐stimulating factor response in myeloid leukaemia cell lines

“…Although HL60 (Ruscetti et al, 1981) and KGI cells (Lusis & Koeffler, 1980) initially proliferated in response to CSF, they lost this property with serial culture (Taetle et al, 1983a;Ruscetti et al, 1981). Recent studies confirm that these cell lines proliferate in response to purified, recombinant (GM)-CSF (Tomonaga et al, 1986). Our HL60 cells never showed CSF responses in liquid or colony culture.…”

Interactions of dimethyl sulfoxide and granulocyte‐macrophage colony‐stimulating factors on the growth and maturation of HL‐60 cells

“…The HL-60 cell line, established from the blood leukocytes of a patient with acute progranulocytic leukemia, has been widely used to study factors influencing the growth and maturation of myeloid cells (Gallagher et al, 1979;KoeMer and Golde, 1980;Olsson and Olofsson, 1981;Taketazu et al, 1984). Despite these efforts, questions remain concerning the relative contributions of the various granulocytelmacrophage colony-stimulating factors (M-, GM-, G-CSFs) (Ruscetti et al, 1981;Metcalf, 1983;Tomonaga et al, 1986) and autocrine mediators other than CSFs Perkins et al, 1984;Heil and Chiao, 1985) to HL-60 growth, and their interactions, if any, with chemical inducers of maturation (Elias et al, 1980). This is particularly true of highly passaged populations, which, in contrast to their earlier counterparts, are highly proliferative, induction resistant, and capable of growing autonomously even at very low cell densities (Gallagher et al, 1979;Brennan et al, 1981).…”