title: Selective production of interferon-alpha subtypes by cultured peripheral blood mononuclear cells and lymphoblastoid cell lines.
abstract: The biological significance of the existence of multiple interferon-alpha (IFN-alpha) subtypes is unknown but may represent a finely tuned mechanism whereby different subtypes are produced in response to different stimuli. To investigate the expression of individual IFN-alpha subtypes, polyclonal antipeptide antisera designed to react with all IFN-alpha subtypes, or with a particular subtype, IFN-alpha 2 or IFN-alpha 4, have been produced. In this study we demonstrate the utility of these antisera for the detection, using indirect immunofluorescence staining, of intracellular IFN-alpha produced by human peripheral blood mononuclear cells (PBMC) and lymphoblastoid cells. Secreted IFN-alpha was also investigated by bioassay and a sandwich radioimmunoassay (RIA), using two monoclonal antibodies (mAb) and specific for IFN-alpha 4. The PBMC were shown to produce IFN reactive with all three polyclonal antisera, after stimulation with Sendai virus. The lymphoblastoid cells also produced IFN, including IFN-alpha 2, but IFN-alpha 4 was not detected either intracellularly, by immunofluorescence, or in the medium, by sandwich RIA. The immunofluorescence studies also demonstrate that in the absence of viral stimulation IFN-alpha is found in the cytoplasm of PBMC and lymphoblastoid cells but not secreted in detectable levels. The finding that two lymphoblastoid cell lines do not produce the subtype IFN-alpha 4 raises important questions as to whether other cell lines and cell types produce IFN-alpha subtypes selectively, and whether individual IFN-alpha subtypes have different roles in human physiology and pathology.
authors.affiliation: Centre for Molecular Biology and Medicine, Monash University, Clayton, Victoria, Australia.
authors.forename: A L
authors.initials: AL
authors.lastname: Greenway
authors.forename: M L
authors.initials: ML
authors.lastname: Overall
authors.forename: N
authors.initials: N
authors.lastname: Sattayasai
authors.forename: M J
authors.initials: MJ
authors.lastname: Rowley
authors.forename: P J
authors.initials: PJ
authors.lastname: Hertzog
authors.forename: G L
authors.initials: GL
authors.lastname: McMullen
authors.forename: B F
authors.initials: BF
authors.lastname: Cheetham
authors.forename: S
authors.initials: S
authors.lastname: Marzuki
mesh_terms: D001681:Biological Assay; D015153:Blotting, Western; D002461:Cell Line, Transformed; D002472:Cell Transformation, Viral / Q000276:immunology; D002478:Cells, Cultured; D005455:Fluorescent Antibody Technique; D006801:Humans; D016898:Interferon-alpha / Q000096:biosynthesis*; D007963:Leukocytes, Mononuclear / Q000276:immunology*; D008214:Lymphocytes / Q000276:immunology
chemical_list: D016898:Interferon-alpha