title: Synergistic effects of nerve growth factor and granulocyte-macrophage colony-stimulating factor on human basophilic cell differentiation.
abstract: We have recently shown that nerve growth factor (NGF) promotes human granulopoiesis, specifically augmenting basophilic cell differentiation observed in methylcellulose hematopoietic colony assays of human peripheral blood. Because the NGF effect was seen in the presence of conditioned medium derived from a human T-cell line (Mo-CM) containing granulocyte-macrophage colony-stimulating factor (GM-CSF), we examined interactions of purified NGF and recombinant human GM-CSF (rhGM-CSF) on granulocyte growth and differentiation. rhGM-CSF stimulated a dose-dependent increase in methylcellulose colony growth at concentrations between 0.1 U/mL and 10 U/mL, and in the presence of NGF at 500 ng/mL this effect was enhanced. The number of basophilic cell colony-forming units (CFU-Baso) and histamine-positive colonies increased synergistically when NGF was added to rhGM-CSF. Furthermore, because Mo-CM acts with sodium butyrate to promote basophilic differentiation of alkaline-passaged myeloid leukemia cells, HL-60, we also examined the interaction of NGF and Mo-CM or rhGM-CSF using this assay. In the presence of NGF, Mo-CM at concentrations of 0.5% to 20% vol/vol, and rhGM-CSF at concentrations of 0.1 U/mL to 100 U/mL synergistically increased histamine production by butyrate-induced, alkaline-passaged HL-60 cells; this was associated with the appearance of metachromatic, tryptase-negative, IgE receptor-positive cells. The effects of rhGM-CSF or Mo-CM were completely abrogated by a specific anti-rhGM-CSF neutralizing antibody in methylcellulose, with or without NGF; the NGF synergy with rhGM-CSF in the HL-60 assay was also inhibited by either anti-rhGM-CSF or anti-NGF antibody. These studies support the notion that differentiation in the basophilic lineage may be enhanced by NGF acting to increase the number of GM-CSF-responsive basophilic cell progenitors.
authors.affiliation: Department of Medicine, McMaster University, Hamilton, Ontario, Canada.
authors.forename: T
authors.initials: T
authors.lastname: Tsuda
authors.forename: D
authors.initials: D
authors.lastname: Wong
authors.forename: J
authors.initials: J
authors.lastname: Dolovich
authors.forename: J
authors.initials: J
authors.lastname: Bienenstock
authors.forename: J
authors.initials: J
authors.lastname: Marshall
authors.forename: J A
authors.initials: JA
authors.lastname: Denburg
mesh_terms: D001491:Basophils / Q000166:cytology*; D002454:Cell Differentiation / Q000187:drug effects; D003470:Culture Media; D004357:Drug Synergism; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000008:administration & dosage*; D006632:Histamine / Q000378:metabolism; D006801:Humans; D007158:Immunologic Techniques; D066298:In Vitro Techniques; D007951:Leukemia, Myeloid / Q000473:pathology; D015458:Leukemia, T-Cell / Q000473:pathology; D009414:Nerve Growth Factors / Q000008:administration & dosage*; D011994:Recombinant Proteins; D014407:Tumor Cells, Cultured
chemical_list: D003470:Culture Media; D009414:Nerve Growth Factors; D011994:Recombinant Proteins; D006632:Histamine; D016178:Granulocyte-Macrophage Colony-Stimulating Factor