Characterization of human blood dendritic cell subsets

MacDonald, Kelli P. A.; Munster, David J.; Clark, Georgina J.; Dzionek, Andrzej; Schmitz, Juergen; Hart, Derek N. J.

doi:10.1182/blood-2001-11-0097

Cited by 660 publications

(674 citation statements)

References 33 publications

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“…Classically, DCs have been defined as two main subsets: Lin2HLA-DR1 CD11c1 CD1232 mDC and the Lin2HLA-DR1 CD11c2 CD1231 pDC (30,(32)(33)(34). More recently, MacDonald et al distinguished five cell subsets within Lin2HLA-DR1 cells: Lin2HLA-DR1 CD1231 pDC, CD341 hematopoietic stem cells, and three subsets of CD11c1 mDCs expressing CD16, CD1c (BDCA-1), or CD141 (BDCA-3) similar to what we describe here (18). However, the authors had to use a cell purification technique to remove all the unwanted cells, prior to the staining.…”

Section: Discussionmentioning

confidence: 99%

“…The identification of specialized lymphocyte subsets such as naïve, memory, or cytotoxic T lymphocytes or monocyte subsets has considerably helped our understanding of immunopathogenesis during HIV and SIV infection (16). Moreover, the polychromatic flow cytometry technique has become increasingly useful in identifying rare subsets of cells such as DC, where a minimum of eight fluorescent parameters, in addition to the physical parameters [forward scatter (FSC) and side scatter (SSC)], are ideal to distinguish five nonoverlapping DC subsets simultaneously (17,18).…”

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confidence: 99%

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Evaluation of a 12‐color flow cytometry panel to study lymphocyte, monocyte, and dendritic cell subsets in humans

et al. 2010

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Monitoring changes in human immune cell populations such as lymphocytes, monocytes, and dendritic cells (DCs) during infectious diseases like human immunodeficiency virus (HIV) is crucial. However, difficulties to identify rare or heterogeneous cell populations can be limiting. For example, to accurately measure DC subsets, eight flow cytometry parameters are ideal. The aim of this work was to analyze the phenotype of human lymphocyte, monocyte, and DC subsets using a single 12-color flow cytometry panel. After erythrocyte lysis, blood from healthy human volunteers was washed and labeled with a cocktail of 12 antibodies. Samples were analyzed on a Becton-Dickinson FACSAria TM equipped with three lasers. Data were compared with lineage-specific panels using 5-8 Ab combinations per lineage. Acquired data were analyzed using FlowJo software. Our 12-color panel allows for the identification of the following major subsets of circulating cells in a single tube: CD41 and CD81 T lymphocytes, B lymphocytes, NK cells, NKT cells, monocyte subsets (CD14 and/or CD16), and five nonoverlapping HLA-DR1Lin2 subsets: CD341 hematopoietic stem cells, CD1231 plasmacytoid DC, and three subsets of CD11c1 myeloid DC expressing either CD16, CD1c (BDCA-1), or CD141 (BDCA-3). We have developed a single flow cytometry panel that allows for simultaneous detection of the lymphocyte and monocyte cell populations and all known DC subsets. Studying these major players of the immune system in one single panel may give us a broader view of the immune response during HIV infection and the ability to better define the role of individual cell types in Acquired Immune Deficiency Syndrome (AIDS) pathogenesis. ' 2010 International Society for Advancement of Cytometry

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Section: Discussionmentioning

confidence: 99%

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confidence: 99%

Evaluation of a 12‐color flow cytometry panel to study lymphocyte, monocyte, and dendritic cell subsets in humans

et al. 2010

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“…Four subtypes have been identified including the CD304 1 (BDCA4) pDCs and the cDCs that include the CD141 1 (BDCA3) and CD1b/c 1 (BDCA1) subsets. 12 A fourth subset that is CD16 1 has been suggested to be a subset of both DC 12 and monocytes, 13 but is now considered to be a monocyte in a recent reclassification of the monocyte and DC subsets in human blood. 14 The CD141 1 DCs have been suggested to be equivalent to the mouse CD8 1 DCs sharing a similar transcriptional signature 15 and expression of Necl2, CLEC9A and XCR1.…”

Section: Introductionmentioning

confidence: 99%

The equivalents of human blood and spleen dendritic cell subtypes can be generated in vitro from human CD34+ stem cells in the presence of fms-like tyrosine kinase 3 ligand and thrombopoietin

et al. 2012

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Dendritic cells (DCs) are immune cells specialized to capture, process and present antigen to T cells in order to initiate an appropriate adaptive immune response. The study of mouse DC has revealed a heterogeneous population of cells that differ in their development, surface phenotype and function. The study of human blood and spleen has shown the presence of two subsets of conventional DC including the CD1b/c 1 and CD141 1 CLEC9A 1 conventional DC (cDC) and a plasmacytoid DC (pDC) that is CD304 1 CD123 1 . Studies on these subpopulations have revealed phenotypic and functional differences that are similar to those described in the mouse. In this study, the three DC subsets have been generated in vitro from human CD34 1 precursors in the presence of fms-like tyrosine kinase 3 ligand (Flt3L) and thrombopoietin (TPO). The DC subsets so generated, including the CD1b/c 1 and CLEC9A 1 cDCs and CD123 1 pDCs, were largely similar to their blood and spleen counterparts with respect to surface phenotype, toll-like receptor and transcription factor expression, capacity to stimulate T cells, cytokine secretion and cross-presentation of antigens. This system may be utilized to study aspects of DC development and function not possible in vivo.

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“…At least two subsets of DCs were originally identified in peripheral blood, including myeloid DCs (mDCs) and plasmacytoid DCs (pDCs). [3][4][5][6] The mDCs express CD11c and CD1c (BDCA-1) antigens. In contrast, pDCs express CD123, BDCA-2 and BDCA-4 antigens.…”

Section: Introductionmentioning

confidence: 99%

IFN-α production by human mononuclear cells infected with varicella-zoster virus through TLR9-dependent and -independent pathways

Huang

Kuo

et al. 2011

Cell Mol Immunol

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Understanding the defense mechanisms of the host of an organism is important for infection control. In previous studies, we demonstrated that interferon-a (IFN-a), but not IL-12, was produced by human peripheral blood mononuclear cells infected with varicella-zoster virus (VZV). Here, we investigated what kind of cell(s) and which signal molecule(s) are involved in IFN-a production. Using cell isolation and ELISA, we found that plasmacytoid dendritic cells (pDCs) were responsible for IFN-a production during VZV infection. We also found that Toll-like receptor 9 (TLR9) was involved in VZV-induced IFN-a production because inhibitory CpG oligodeoxynucleotide inhibited IFN-a production. UV-inactivated VZV-induced IFN-a production was lower than that of active VZV, indicating another TLR9-independent pathway. Further studies demonstrated that double-stranded RNA-dependent protein kinase, but not DNA-dependent protein kinase was involved in VZV-induced IFN-a production. Together, these results suggest that pDCs play an important role in IFN-a production during VZV infection through TLR9-dependent and -independent pathways.

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Characterization of human blood dendritic cell subsets

Cited by 660 publications

References 33 publications

Evaluation of a 12‐color flow cytometry panel to study lymphocyte, monocyte, and dendritic cell subsets in humans

Evaluation of a 12‐color flow cytometry panel to study lymphocyte, monocyte, and dendritic cell subsets in humans

The equivalents of human blood and spleen dendritic cell subtypes can be generated in vitro from human CD34+ stem cells in the presence of fms-like tyrosine kinase 3 ligand and thrombopoietin

IFN-α production by human mononuclear cells infected with varicella-zoster virus through TLR9-dependent and -independent pathways

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