Selective production of interferon-alpha subtypes by cultured peripheral blood mononuclear cells and lymphoblastoid cell lines

A. L. Greenway, M. L. Overall, N. Sattayasai, M. J. Rowley, P. J. Hertzog, G. L. McMullen, B. F. Cheetham, S. Marzuki

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17 Citations (Scopus)

Abstract

The biological significance of the existence of multiple interferon-alpha (IFN-α) 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-α subtypes, polyclonal antipeptide antisera designed to react with all IFN-α subtypes, or with a particular subtype, IFN-α2 or IFN-α4, have been produced. In this study we demonstrate the utility of these antisera for the detection, using indirect immunofluorescence staining, of intracellular IFN-α produced by human peripheral blood mononuclear cells (PBMC) and lymphoblastoid cells. Secreted IFN-α was also investigated by bio-assay and a sandwich radioimmunoassay (RIA), using two monoclonal antibodies (mAb) and specific for IFN-α4. 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-α2, but IFN-α4 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-α 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-α4 raises important questions as to whether other cell lines and cell types produce IFN-α subtypes selectively, and whether individual IFN-α subtypes have different roles in human physiology and pathology.

Original languageEnglish
Pages (from-to)182-188
Number of pages7
JournalImmunology
Volume75
Issue number1
Publication statusPublished - 1 Jan 1992

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