Dendritic Cell Research

Human dendritic cells

Human dendritic cell (DC) subsets can be characterized by a number of surface and intracellular markers through flow cytometry.

See Table 2 for key markers for phenotypic characterization of various human DC subsets; intracellular markers are indicated in bold. BD Life Sciences has a broad portfolio of antibody reagents to most of these specificities in multiple formats to allow flexibility in panel design and downstream analysis.


All human DC subsets are identified as lineage (CD3 CD19 CD20 CD56 CD141) and CD45+ MHCII (HLA-DR)+ CD11c2.

 

Table 2. Human DC subsets

DC Subset Key Markers
Primary Markers Additional Positive Markers Additional Negative Markers Transcription Factors
Plasmacytoid DCs CD123 (IL-3Rα)high
CD303 (BDCA2/CLEC4C)+
CD304 (Neuropilin-1/BDCA4)+
CD85g (ILT7)+
CD11clow
CD11b, CD14
CD2±, CD4+, CD45RA+
CD141 (BDCA3)low
CD272 (BTLA)+
CD366 (TIM-3)±
CD367 (DCIR/CLEC4A)+
CD371 (CLEC12A)±

TLR7high, TLR9high

FLT3 (CD135)+
GM-CSFR (CD116)+

CCR5+, CXCR4+
CD1a, CD1c (BDCA1)
CD16 (FcγRIII)
CD172a (Sirp-α)
CD207 (Langerin)
CD324 (E-Cadherin)
CD326 (EpCAM)
CD369 (Dectin-1/CLEC7A)
CD370 (CLEC9A/DNGR1)
CLEC6A (Dectin-2)

XCR1
IRF7+, IRF8+
SpiB+
CD1c+ Myeloid DCs CD1c (BDCA1)+
CD172a (Sirp-α)+
CLEC6A (Dectin-2)+
CD11b+/low, CD11c+
CX3CR1+
CD14low/–
CD4+, CD13+, CD26low, CD33+
CD45RO+, CD141 (BDCA3)±
CD272(BTLA)+
CD366 (TIM-3)+
CD367 (DCIR/CLEC4A)+
CD369 (Dectin-1/CLEC7A)+
CD371 (CLEC12A)+
CD1a+ [skin and intestine]
CD141(BDCA3)+ [intestine]

TLR3low, TLR4low, TLR8+, TLR10low

FLT3 (CD135)+
GM-CSFR (CD116)+
CD1a, CD16 (FcγRIII)
CD123 (IL-3Rα)
CD207 (Langerin)
CD304 (Neuropilin-1/BDCA4), CD324 (E-Cadherin)
CD326 (EpCAM)
CD370 (CLEC9A/DNGR1)
ESAM

XCR1
IRF4+
CD141+ Myeloid DCs CD141 (BDCA3)high
CD370 (CLEC9A/DNGR1)+
NECL2 (CADM1)+
CD11c+/low
CD14
CD4+, CD11blow
CD26+, CD162high
CD205 (DEC-205)high
CD272 (BTLA)high
CD367 (DCIR/CLEC4A)+
CD366 (TIM-3)+
CD369 (Dectin-1/CLEC7A)+
CD371 (CLEC12A)+

TLR3+, TLR8+

FLT3 (CD135)+
GM-CSFR (CD116)+

XCR1+
CD1a, CD1c (BDCA1), CD16 (FcγRIII)
CD172a (Sirp–α)
CD207 (Langerin)
CD304 (Neuropilin-1/BDCA4)
CD324 (E-Cadherin)
CD326 (EpCAM)
IRF8+
BATF3+
Langerhans Cells CD207 (Langerin)+
CD324 (E-Cadherin)+
CD326 (EpCAM)+
CD11blow, CD11c+
CD14
CD1ahigh, CD1c (BDCA1)+
CD36+
CD172a (Sirp-α)+
CD369 (Dectin-1/CLEC7A)+, CD371 (CLEC12A)+
CLEC6A (Dectin-2)+

TLR1+, TLR2+, TLR3low, TLR6+
CD304 (Neuropilin-1/BDCA4)
CD367 (DCIR/CLEC4A)
XCR1
 
CD1a+ Dermal DCs CD1a+
CD64 (FcγRI)+
CD366 (TIM-3)+
CD11b+, CD11c+
CD14
CD1c (BDCA1)+
CD172a (Sirp-α)+
CD367 (DCIR/CLEC4A)+
CD369 (Dectin-1/CLEC7A)+
CLEC6A (Dectin-2)+

TLR1–3+, TLR6+, TLR7+, TLR10+
CD207 (Langerin)
CD209 (DC-SIGN/CLEC4L)
CD324 (E-Cadherin)
CD326 (EpCAM)
 
CD14+ Dermal DCs CD14+
CD209 (DC-SIGN/CLEC4L)+
CD11b+, CD11c+
CD1c (BDCA1)+
CD172a (Sirp-α)+
CD367 (DCIR/CLEC4A)+
CD369 (Dectin-1/CLEC7A)+
CLEC6A (Dectin-2)+

TLR1–3+, TLR6, TLR7+
CSF–1R (CD115)+
CD1a
CD207 (Langerin)
CD324 (E-Cadherin)
CD326 (EpCAM)
CD366 (TIM-3)
 

Inflammatory DCs

(Monocyte-Derived DCs)

CD16 (FcγRIII) +
CD64 (FcγRI)+
CD1a+
CD1c (BDCA1)+
CD11b+, CD11c+
CD14±
CD172a (Sirp-α)+
CD206 (MR/CLECL13D)+
CD209 (DC-SIGN/CLEC4L)+
CD367 (DCIR/CLEC4A)+
CD369 (Dectin-1/CLEC7A)+
CD371 (CLEC12A)+
CLEC6A (Dectin-2)+

TLR3low, TLR4+, TLR7low, TLR8+

CCR2 (CD192)+
CD207 (Langerin)
CD324 (E-Cadherin)
CD326 (EpCAM)
 

1CD14 is negative or low on all DC subsets except for CD14+ dermal DCs and inflammatory DCs.

2CD11c is positive on all DC subsets except for plasmacytoid DCs where expression has been reported as low or negative.

TLR7, 8 and 9 are endosomal and require intracellular staining.

The following examples on peripheral blood illustrate DC subset identification based on available instruments and laser choices. Additional catalog reagents as well as BD OptiBuild™ custom reagents are available for further characterization of various human DC subsets.

Analysis of human DC populations on a BD LSRFortessa™ X-20 cell analyzer
Analysis of human DC populations on a BD LSRFortessa X-20 cell analyzer

Analysis of human DC populations on a three-laser BD FACSVerse™ flow cytometer
Analysis of human DC populations on a three-laser BD FACSVerse flow cytometer


Bibliography

  1. Merad M, Sathe P, Helft J, Miller J, Mortha A. The dendritic cell lineage: ontogeny and function of dendritic cells and their subsets in the steady state and the inflamed setting. Ann Rev Immunol. 2013;31:563-604.
  2. Schraml BU, Reis e Sousa C. Defining dendritic cells. Curr Opin Immunol. 2015;32:13-20.
  3. Collin M, McGovern N, Haniffa M. Human dendritic cell subsets. Immunology. 2013;140:22-30.
  4. O'Keeffe M, Mok WH, Radford KJ. Human dendritic cell subsets and function in health and disease. Cell Mol Life Sci. 2015;72:4309-4325.
  5. Apostolopoulos V, Thalhammer T, Tzakos AG, Stojanovska L. Targeting antigens to dendritic cell receptors for vaccine development. J Drug Deliv. 2013;2013:869718.
  6. Cohn L, Delamarre L. Dendritic cell-targeted vaccines. Front Immunol. 2014;5:255.
  7. Delamarre L, Mellman I. Harnessing dendritic cells for immunotherapy. Sem Immunol. 2011;23:2-11.
  8. Breton G, Lee J, Liu K, Nussenzweig MC. Defining human dendritic cell progenitors by multiparametric flow cytometry. Nat Protoc. 2015;10:1407-1422.
  9. Poltorak MP, Schraml BU. Fate mapping of dendritic cells. Front Immunol. 2015;6:199.
  10. Swiecki M, Colonna M. The multifaceted biology of plasmacytoid dendritic cells. Nat Rev Immunol. 2015;15:471-485.
  11. Murphy TL, Grajales-Reyes GE, Wu X, et al. Transcriptional control of dendritic cell development. Annu Rev Immunol. 2016;34:93-119.
  12. Durand M, Segura E. The known unknowns of the human dendritic cell network. Front Immunol. 2015;6:129.
  13. Dutertre CA, Wang LF, Ginhoux F. Aligning bona fide dendritic cell populations across species. Cell Immunol. 2014;291:3-10.
  14. Schlitzer A, Ginhoux F. Organization of the mouse and human DC network. Curr Opin Immunol. 2014;26:90-99.
  15. Reis e Sousa C. Activation of dendritic cells: translating innate into adaptive immunity. Curr Opin Immunol. 2004;16:21-25.
  16. Malissen B, Tamoutounour S, Henri S. The origins and functions of dendritic cells and macrophages in the skin. Nat Rev Immunol. 2014;14:417-428.
  17. Henri S, Poulin LF, Tamoutounour S, et al. CD207+ CD103+ dermal dendritic cells cross-present keratinocyte-derived antigens irrespective of the presence of Langerhans cells. J Exp Med. 2010;207:189-206.
  18. Klechevsky E, Morita R, Liu M, et al. Functional specializations of human epidermal Langerhans cells and CD14+ dermal dendritic cells. Immunity. 2008;29:497-510.
  19. Autenrieth SE, Grimm S, Rittig SM, et al. Profiling of primary peripheral blood- and monocyte-derived dendritic cells using monoclonal antibodies from the HLDA10 Workshop in Wollongong, Australia. Clin Transl Immunology. 2015;4:e50.
  20. Ohradanova-Repic A, Machacek C, Fischer MB, Stockinger H. Differentiation of human monocytes and derived subsets of macrophages and dendritic cells by the HLDA10 monoclonal antibody panel. Clin Transl Immunology. 2016;5:e55.
  21. Clark GJ, Kupresanin F, Fromm PD, et al. New insights into the phenotype of human dendritic cell populations. Clin Transl Immunology. 2016;5:e61.
  22. Fromm PD, Kupresanin F, Brooks AE, et al. A multi-laboratory comparison of blood dendritic cell populations. Clin Transl Immunology. 2016;5:e68.


For Research Use Only. Not for use in diagnostic or therapeutic procedures.