title: Production of hematopoietic colony-stimulating factors by human natural killer cells.
abstract: We have analyzed the ability of highly purified preparations of human NK cells to produce CSF. NK cells, purified by negative selection from 10-d cultures of PBMC incubated with irradiated B-lymphoblastoid cell lines, were stimulated with rIL-2, FcR(CD16) ligands (particulate immune complexes or anti-CD16 antibodies bound to Sepharose), a combination of CD16 ligands and rIL-2, or the phorbol diester phorbol dibutyrate (PDBu) together with the Ca2+ ionophore A23187. Both rIL-2 and CD16 ligands induce accumulation of GM-CSF mRNA in NK cells and the combined effect of the two stimuli is synergistic. Maximal accumulation of GM-CSF mRNA is observed after PDBu/A23187 stimulation. The participation of contaminant T cells in the observed expression of the GM-CSF gene is excluded because CD16 ligands do not stimulate T cells and CD3 ligands, powerful stimulators of T cells, are inactive on NK cells. Accumulation of CSF-1 mRNA is observed only in NK cells stimulated with both CD16 ligands and rIL-2, whereas accumulation of IL-3 mRNA is observed only in NK cells stimulated with PDBu/A23187. Transcripts of the G-CSF, IL-1 alpha, and IL-1 beta genes were never detected in NK cells in these experiments. The kinetics of accumulation of GM-CSF and CSF-1 mRNA in NK cells stimulated with CD16 ligands and rIL-2 peaked at 2-4 h and was slower than that of TNF and IFN-gamma mRNA, which peak at 1 h. GM-CSF was precipitated from the supernatant fluids of NK cells stimulated with PDBu/A23187 and its biological activity was demonstrated by the ability of the supernatants to sustain proliferation of the TALL-101 cell line or CML blasts. Biological activity of IL-3 and CSF-1 was demonstrable in supernatant fluids of NK cells stimulated with PDBu/A23187 and CD16 ligands/rIL-2, respectively.
authors.affiliation: Wistar Institute of Anatomy and Biology, Philadelphia, Pennsylvania 19104.
authors.forename: M C
authors.initials: MC
authors.lastname: Cuturi
authors.forename: I
authors.initials: I
authors.lastname: Anegón
authors.forename: F
authors.initials: F
authors.lastname: Sherman
authors.forename: R
authors.initials: R
authors.lastname: Loudon
authors.forename: S C
authors.initials: SC
authors.lastname: Clark
authors.forename: B
authors.initials: B
authors.lastname: Perussia
authors.forename: G
authors.initials: G
authors.lastname: Trinchieri
mesh_terms: D000943:Antigens, Differentiation / Q000502:physiology; D000945:Antigens, Differentiation, T-Lymphocyte / Q000502:physiology; D001681:Biological Assay; D015152:Blotting, Northern; D017252:CD3 Complex; D000001:Calcimycin / Q000494:pharmacology; D002478:Cells, Cultured; D003115:Colony-Stimulating Factors / Q000096:biosynthesis*; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D006133:Growth Substances / Q000096:biosynthesis*; D006801:Humans; D007375:Interleukin-1 / Q000096:biosynthesis; D007376:Interleukin-2 / Q000494:pharmacology; D007377:Interleukin-3 / Q000096:biosynthesis*; D007694:Killer Cells, Natural / Q000502:physiology*; D008024:Ligands; D010703:Phorbol Esters / Q000494:pharmacology; D011233:Precipitin Tests; D012333:RNA, Messenger / Q000235:genetics; D011948:Receptors, Antigen, T-Cell / Q000502:physiology; D011961:Receptors, Fc / Q000502:physiology; D017452:Receptors, IgG
chemical_list: D000943:Antigens, Differentiation; D000945:Antigens, Differentiation, T-Lymphocyte; D017252:CD3 Complex; D003115:Colony-Stimulating Factors; D006133:Growth Substances; D007375:Interleukin-1; D007376:Interleukin-2; D007377:Interleukin-3; D008024:Ligands; D010703:Phorbol Esters; D012333:RNA, Messenger; D011948:Receptors, Antigen, T-Cell; D011961:Receptors, Fc; D017452:Receptors, IgG; D000001:Calcimycin; D016178:Granulocyte-Macrophage Colony-Stimulating Factor