Identification of plasmacytoid pre-dendritic cells by one-color flow cytometry for phenotype screening

Plasmacytoid pre-dendritic cells (pDCs) are able to prime and polarize naive T-cells, while also having an important effector function in antiviral immunity through the rapid and robust production of interferon-alpha. The main setback of pDCs investigation it; the rarity and ex vivo fragility of these cells. Relative simple, reliable, and accurate methods for phenotypic analysis and functional studies of pDCs without isolation would be a great deal of interest. Fresh whole blood samples were analyzed by two-color and one-color flow cytometric pDC-identification assays. The changes in the surface expression of CD62L and HLA-DQ on pDCs in whole blood samples after 24-h treatment with imiquimod, a toll-like receptor 7 agonist, were analyzed. Our data demonstrate that the identification of pDCs in peripheral blood samples can be achieved by using only one fluorescent channel for blood dendritic cell antigen (BDCA)-4 staining combined with the light scatter parameters, thus leaving the other channels open for further phenotypic and/or functional analysis. Recently, several lines of evidence supported the involvement of pDCs in the development of several human diseases, so our new one-color identification approach may provide a useful tool for investigation of the pathomechanism of the relevant diseases by using common, 2-laser benchtop, cytometers. (c) 2008 international Society for Analytical Cytology.