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

INTRODUCTION: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. METHODS: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. RESULTS: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 observed by IFN-alpha above 100 IU/ml. DISCUSSION:This assay may be useful for examining IFNAR2 in various leukocyte subsets, separately, as well as providing a rapid and easy method for monitoring expression of type I IFN receptors.