A flow cytometric method for determination of the interferon receptor IFNAR2 subunit in peripheral blood leukocyte subsets

Journal of Pharmacological and Toxicological Methods
Shiro TochizawaToyoki Mori

Abstract

It is expected that expression levels in the peripheral blood mononuclear cells (PBMC) of IFNAR2, a subunit of the interferon (IFN) receptor, may be a marker for predicting IFN response. In the present study, we have established a rapid and convenient method for assaying IFNAR2, using flow cytometry. Fifty microliters of whole blood from healthy volunteers was treated with an anti-IFNAR2 antibody and stained with a Fluorescein isothiocyanate (FITC)-conjugated secondary antibody. In addition, the cells were stained with subset-specific antibodies conjugated with phycoerythrin (PE) and PE covalently linked to cyanin 5 at the same time. The mean FITC-fluorescence intensities were analyzed separately by gating on subset-specific regions. IFNAR2 was detected in most lymphocytes, monocytes, and granulocytes, although IFNAR2 expression was higher in the monocytes and granulocytes than in the lymphocytes. The intra- and interdaily variations of IFNAR2 in lymphocytes, monocytes, and granulocytes were small. Among the lymphocyte subsets, IFNAR2 showed high expression in natural killer (NK) cells and low expression in T lymphocytes. The effect of IFN-alpha on IFNAR2 expression was examined in vitro. A down-regulation of IFNAR2 was observe...Continue Reading

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Citations

Mar 30, 2011·Journal of Clinical Immunology·Fanli MengKai Wang
Sep 7, 2006·Liver International : Official Journal of the International Association for the Study of the Liver·Hirohisa YanoMasamichi Kojiro
Jun 6, 2006·Biochemical and Biophysical Research Communications·Shiro TochizawaToyoki Mori
Sep 5, 2019·Experimental Biology and Medicine·Steven R GoodmanDavid Kakhniashvili
Mar 31, 2017·Journal of Molecular Medicine : Official Organ of the Gesellschaft Deutscher Naturforscher Und Ärzte·Romy StraußRobert Biesen

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