Magyarics Zoltan, Csillag Aniko, Pazmandi Kitti, Rajnavolgyi Eva, Bacsi Attila
Institute of Immunology, Medical and Health Science Center, Faculty of Medicine, University of Debrecen, 98 Nagyerdei Blvd., Debrecen H-4012, Hungary.
Cytometry A. 2008 Mar;73(3):254-8. doi: 10.1002/cyto.a.20529.
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 is 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.
浆细胞样前体树突状细胞（pDC）能够启动并极化初始T细胞，同时通过快速大量产生α干扰素在抗病毒免疫中发挥重要的效应功能。pDC研究的主要障碍是这些细胞稀少且离体时脆弱。相对简单、可靠且准确的不进行分离的pDC表型分析和功能研究方法将备受关注。通过双色和单色流式细胞术pDC鉴定分析新鲜全血样本。分析了用Toll样受体7激动剂咪喹莫特处理24小时后全血样本中pDC表面CD62L和HLA-DQ表达的变化。我们的数据表明，仅使用一个荧光通道进行血树突状细胞抗原（BDCA）-4染色并结合光散射参数，就能在外周血样本中鉴定pDC，从而使其他通道可用于进一步的表型和/或功能分析。最近，几条证据支持pDC参与多种人类疾病的发生发展，因此我们新的单色鉴定方法可能为利用常见的双激光台式细胞仪研究相关疾病的发病机制提供有用工具。
