Dendritic cell markers

Dendritic cells play a crucial role in activation, regulation and polarization of the immune responses.

These antigen presenting cells are essential for both immune reactions and for immune tolerance, and they integrate innate and adaptive immunity. Dendritic cells differentiate from bone marrow-residing progenitors, and circulate in the blood, untill they enter peripheral tissues to play their part.

DC.jpgDendritic cell life in the peripheral tissues lasts only for days to weeks, thus they must be continuously replaced by new ones.  There are four main dendritic cell types and some other dendritic cell-relative cell types.  A simplified view on these populations is following: Classical myeloid dendritic cells type 1 (cDC1) are crucial for induction of cytotoxic T cell immunity against intracellular pathogens and cancer. Classical myeloid dendritic cells type 2 (cDC2) play an important role in detection of extracellular pathogens and presentation of their antigens. Plasmacytoid dendritic cells (pDC) resemble morphologically to the plasma cells, and their main function seems to be rapid production of antiviral interferons. Monocyte-derived dendritic cells (Mo-DC) develope from some monocytes during inflammation. Similar origin have epithelial Langerhans cells.  Sometimes are among dendritic cells sensu lato counted also non-classical monocytes. 

Fig. 1: Among other functions, dendritic cells also play an important role in the process of allergic hypersensitivity.

Table of dendritic cell markers:

Dendritic cell types

Markers (antibody clone: formats)

Classical myeloid
dendritic cells type 1

CD141 (M80 coming soon)
CD26   (BA5b: Purified, PB, FITC, PE, APC)
CD226 (11A8 coming soon)
CD103 (Ber-ACT8:  Purified, FITC, PE, APC)

Classical myeloid
dendritic cells type 2

CD11b (ICRF44: Purified, biotin, PB, FITC, AF 488, PE, APC, PC7, AC7, AF 700)
             (MEM-174: Purified, biotin, FITC, PE, APC)
             (MEM-171: Purified)
CD11c (BU15: Purified, biotin, PB, FITC, AF 488, PE, APC, PerCP, PerC5.5, PC7, AC7)
CD1c   (L161: Purified, PE, APC)
CD2     (LT2: Purified, PB, FITC, PE, APC)
            (TS1/8: Purified, PB, PE, PC7, AC7)
            (MEM-65:  Purified, biotin, PE)
CD163 (GHI/61: Purified, PE, APC, PerC5.5, PC7)

dendritic cells (pDC)

CD123 (6H6: Purified, biotin, FITC, PE, APC, PC5, PerCPC5.5, PC7)
CD45RA (MEM-56:  Purified, biotin, PB, FITC, PO, PDL 594, PE, APC, PerCP, PerC5.5, PC7, AC7)
CD85k (Purified, more coming soon)

dendritic cells

CD209 (UW60.1: Purified, PE, APC, AF 647)
CD206 (15-2: Purified, FITC, PE, APC, AF647, PerC5.5, PC7)
CD1a  (SK9: Purified, FITC, PE, APC)
            (HI149: Purified, FITC, PE, APC)
CD1c   (L161: Purified, PE, APC)
CD11c (BU15: Purified, biotin, PB, FITC, AF 488, PE, APC, PerCP, PerC5.5, PC7, AC7)

Langerhans cells

CD1a  (SK9: Purified, FITC, PE, APC)
            (HI149: Purified, FITC, PE, APC)
CD326 (323/A3: Purified, FITC, AF 488, PE, APC, AF 647, PerC5.5, PC7)
            (VU-1D9: Purified, FITC, PE, APC, AF 647, PerC5.5, PC7)

Purified = unconjugated antibody, PB = Pacific Blue™,  PO = Pacific Orange™,  PDL = PE-DyLight®, AF = Alexa Fluor®, PerC5.5 = PerCP-Cy™5.5, PC5 = PE-Cy™5, PC7 = PE-Cy™7, AC7 = APC- Cy™7
Markers shared with other cell types make flow cytometric identification of so rare cells, like the dendritic cells are, a bit difficult. For example, CD26 is expressed not only on cDC1, but also on lymphocytes and NK cells. CD11b is expressed not only on cDC2, but also on monocytes, granulocytes, NK cells and some T and B cell subsets. Similar situation is with most dendritic cell markers. However, this can be overcome by a lineage cocktail that enables to exclude most undesired populations by identifying a pool of their specific markers under one single fluorochrome.
The Lineage Cocktail FITC (cat no. 6K01-T050) is designed to stain and gate-out human T lymphocytes (CD3+), B lymphocytes (CD19+CD20+), NK cells (CD16+CD56+), non-differentiated monocytes (CD14+), neutrophils (CD16+), and eosinophils (CD14+dim). Basophils and a part of dendritic cells (cDC1 and pDC) are lineage cocktail-negative and do not stain using this reagent. Whenever applicable, the lineage negative cells can be simultaneously stained with other monoclonal antibodies conjugated to appropriate fluorochromes. For example, dendritic cells can be distinguished by positive staining with anti HLA-DR, and basophils by positive staining with anti CD203c. Another markers can be applied for lineage-negative cells to identify particular dendritic cell types, namely those for cDC1 and pDC. As Mo-DC and cDC2 express CD14, they can not be negatively stained with this lineage cocktail.
Lineage_FITC.jpgFig. 2: Flow cytometry multicolor surface staining of human peripheral whole blood stained using Lineage Cocktail FITC (20 μl cocktail / 100 μl of peripheral whole blood) and anti-human CD203c (NP4D6) APC antibody (10 μl reagent / 100 μl of peripheral whole blood). The lineage negative CD203c negative population is rich in CD14 negative dendritic cell types (cDC1 and pDC).


Further reading:
Musumeci A, Lutz K, Winheim E, Krug AB: What makes a pDC: Recent advances in understanding plasmacytoid DC development and heterogeneity. Front Immunol. 2019; 10: 1222.
Breton G, Lee J, Liu K, Nussenzweig MC: Defining human dendritic cell progenitors by multiparametric flow cytometry. Nat Protoc. 2015 Sep;10(9):1407-22.
Collin M, Bigley V: Human dendritic cell subsets: An update. Immunology 2018 May;154(1):3-20.
Jardine L, Barge D, Ames-Draycott A, Pagan S, Cookson S, Spickett G, Haniffa M, Collin M, Bigley V: Rapid detection of dendritic cell and monocyte disorders using CD4 as a lineage marker of the human peripheral blood antigen-presenting cell compartment. Front Immunol. 2013 Dec 27;4:495.
Rovati B, Mariucci S, Manzoni M, Bencardino K, Danova M: Flow cytometric detection of circulating dendritic cells in healthy subjects. Eur J Histochem. 2008 Jan-Mar;52(1):45-52.
Carenza C, Franzese S, Calcaterra F, Mavilio D, Della Bella S. Comprehensive phenotyping of dendritic cells in cancer patients by flow cytometry. Cytometry A. 2021 Mar;99(3):218-230.

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