title: Membrane oxidative metabolism of human eosinophilic cell line EoL-1 in response to phorbol diester and formyl peptide: synergistic augmentation by interferon-gamma and tumor necrosis factor.
abstract: Human eosinophilic cell line EoL-1 was studied using luminol-dependent chemiluminescence (CL) for the ability to produce a respiratory burst upon stimulation with 12-O-tetradecanoylphorbol-13-acetate (TPA) and N-formyl-L-methionyl-L-leucyl-L-phenylalanine (fMLP). TPA, a potent activator of protein kinase C, induced a biphasic CL response in EoL-1. Treatment of EoL-1 with interferon-gamma (IFN-gamma) for 5 days dramatically enhanced TPA-inducible CL, and IFN-alpha A had a similar effect. Neither IFN-gamma nor IFN-alpha A strongly inhibited EoL-1 cell growth. Tumor necrosis factor (TNF) also enhanced TPA-inducible CL response of EoL-1 and, furthermore, was quite inhibitory to EoL-1 cell growth. The effects of IFN-gamma and TNF were synergistic, whereas those of IFN-alpha A and TNF were additive. Superoxide dismutase completely abrogated TPA-induced CL, but sodium azide suppressed only the late phase of CL. EoL-1 pretreated with IFN-gamma, IFN-alpha A, or TNF also became capable of producing CL response to a chemotactic peptide (fMLP). The effects of IFN-gamma and TNF were again synergistic. EoL-1 cells treated with IFN-gamma, IFN-alpha A, or TNF had abundant cytoplasm, but only TNF increased cells having distinct eosinophilic granules. IFN-gamma but not IFN-alpha A enhanced the cytological effect of TNF. It was further demonstrated that treatment of EoL-1 with IFN-gamma and TNF strongly increased the binding sites for phorbol diesters and also dramatically induced the surface expression of fMLP receptors. IFN-gamma had, however, little effect on the number or the ligand-binding affinity of TNF receptors on EoL-1. Thus, EoL-1 may provide a useful experimental model for the study of differentiation and regulation of human eosinophils.
authors.affiliation: Department of Bacteriology, Tohoku University School of Medicine, Sendai, Japan.
authors.forename: O
authors.initials: O
authors.lastname: Yoshie
authors.forename: T
authors.initials: T
authors.lastname: Majima
authors.forename: H
authors.initials: H
authors.lastname: Saito
mesh_terms: D001386:Azides / Q000494:pharmacology; D029742:Caenorhabditis elegans Proteins; D002352:Carrier Proteins; D002460:Cell Line; D004357:Drug Synergism; D004804:Eosinophils / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D007371:Interferon-gamma / Q000494:pharmacology*; D008163:Luminescent Measurements; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology*; D010084:Oxidation-Reduction; D011493:Protein Kinase C; D011956:Receptors, Cell Surface / Q000187:drug effects; D011955:Receptors, Drug / Q000187:drug effects; D044042:Receptors, Formyl Peptide; D011971:Receptors, Immunologic / Q000187:drug effects; D018124:Receptors, Tumor Necrosis Factor; D019810:Sodium Azide; D013482:Superoxide Dismutase / Q000494:pharmacology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*
chemical_list: D001386:Azides; D029742:Caenorhabditis elegans Proteins; D002352:Carrier Proteins; D011956:Receptors, Cell Surface; D011955:Receptors, Drug; D044042:Receptors, Formyl Peptide; D011971:Receptors, Immunologic; D018124:Receptors, Tumor Necrosis Factor; D014409:Tumor Necrosis Factor-alpha; C083141:phorbol ester binding protein; C027776:phorbol ester receptor; D009240:N-Formylmethionine Leucyl-Phenylalanine; D007371:Interferon-gamma; D019810:Sodium Azide; D013482:Superoxide Dismutase; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate