Dendritic Cells Analysis - An 8-color panel for detection of Human blood dendritic cells by flow cytometry
Dendritic cells (DCs) are antigen presenting cells capable of presenting antigen and priming a T cell response. They form a heterogeneous group of cells based on phenotype, location and function. In human blood, DCs represent less than 1% of white blood cells, and can be separated into 2 main cell subsets, namely the myeloid DCs (MDCs) and the plasmacytoid DCs (pDCs). Among the MDCs, 3 distinct cell subsets are identified: CD1c+MDCs (MDC1), CD141+MDCs (MDC2) and CD16+MDCs. In blood, the frequency of DCs is affected in certain pathological conditions such as HIV, diabetes, asthma, chronic viral hepatitis, and graft-versus-host-disease. Thus, the detection and enumeration of different blood DC subsets is important to understand immune regulation in pathological conditions and to guide specific patient treatments. Due to the lack of specific markers for DC definition, the combination of several markers is required to allow their identification. Based on current knowledge in human DC biology, we have evaluated the expression and association of several DC markers to design an optimized 8-color panel for flow cytometry which allows for the detection of all DCs subsets in whole blood samples or peripheral blood mononuclear cells (PBMCs). This panel (CD1c/HLA-DR/Lineage/CD11c/CD16/Clec9A/CD123/CD45) provides an easy and robust assay to study the role of DCs in healthy donors and patient samples.