YILIMART - BDCA-1 Cell Isolation Kit, human

The CD1c (BDCA-1)

⁺

Dendritic Cell Isolation Kit has been developed for the isolation of CD1c (BDCA-1)

⁺

myeloid dendritic cells (MDCs; type-1 MDC or cDC2) from PBMCs. This new format allows for the optional depletion of CD14

⁺

myeloid cells that recently have been shown to possibly contaminate the final DC population.

Background information

The CD1c (BDCA-1) antigen is specifically expressed on dendritic cells, which are CD11c

^high

CD123

^low

and represent the major subset of myeloid dendritic cells in human blood. CD1c (BDCA-1)

⁺

dendritic cells show a monocytoid morphology (fig. 1) and express myeloid markers, such as CD13 and CD33 as well as Fc receptors, such as CD32, CD64, and FcεRI. Furthermore, they were determined to be CD4

⁺

, Lin (CD3, CD16, CD19, CD20, CD56)

^–

, CD2

⁺

, CD45RO

⁺

, CD141 (BDCA-3)

^low

, CD303 (BDCA 2)

^–

, and CD304 (BDCA-4/ Neuropilin-1)

^–

. A minor proportion of CD1c (BDCA-1)

⁺

myeloid dendritic cells expresses CD14 and CD11b. CD1c (BDCA-1) is also found on CD1a

⁺

dendritic cells generated

ex vivo

from monocytes or hematopoietic precursor cells, and on CD1a

⁺

Langerhans cells in skin. Recently, a functionally different population of CD14

⁺

CD1c

⁺

myeloid cells has been reported. To avoid the co-enrichment of such population, an optional removal reagent for CD14

⁺

myeloid cells is included. In blood, apart from myeloid dendritic cells, a subset of small resting B cells expresses CD1c (BDCA-1). For this reason, the CD1c (BDCA-1)

⁺

Dendritic Cell Isolation Kit includes CD19 MicroBeads for depletion of B cells prior to the enrichment of CD1c (BDCA-1)

⁺

myeloid dendritic cells. In order to discriminate the CD1c (BDCA-1)

⁺

from CD141 (BDCA-3)

⁺

myeloid dendritic cells, they have been designated type 1 myeloid dendritic cells (MDC1s) or conventional DC2 cells (cDC2s).

Downstream applications

CD1c (BDCA-1)

⁺

dendritic cells can be used for a variety of applications, for example:

Isolation of CD1c (BDCA-1)⁺ myeloid dendritic cells to examine expression of Toll-like receptors, chemokine receptors, or new antigens, e.g., DCAL-1 and EMR2.
Isolation for studies on dendritic cell activation, migration, cytokine production and T cell polarization, particularly in comparison with monocyte-derived dendritic cells.
Isolation of CD1c⁺ DCs for studies evaluating new checkpoint inhibitors or dissecting their mechanisms of actions in tumor settings.

Columns

For the first magnetic separation (depletion): LD or autoMACS

^®

Columns. For the second magnetic separation (positive selection): MS, LS, or autoMACS Columns.

Figure 1

CD1c (BDCA-1)

⁺

myeloid dendritic cells were isolated from human PBMCs using the CD1c (BDCA-1)

⁺

Dendritic Cell Isolation Kit, an LD Column, two MS Columns, a MidiMACS™ Separator and a MiniMACS™ Separator. Cells were fluorescently stained with CD1c (BDCA-1)-PE, CD14-APC, or CD19-VioBright™ 515. Cell debris and dead cells were excluded from the analysis based on scatter signals and prodipium iodide fluorescence and erythrocytes based on CD45-negativity.

Before depletion	After depletion

Before separation	CD1c (BDCA-1) ⁺ cells

Selected references
1. Dzionek, A. et al. (2000) BDCA-2, BDCA-3, BDCA-4: Three markers for distinct subsets of dendritic cells in human peripheral blood. J. Immunol. 165: 6037-6046
2. Mathan, T. S. M. et al. (2017) Harnessing RNA sequencing for global, unbiased evaluation of two new adjuvants for dendritic-cell immunotherapy. Oncotarget 8(12): 19879-19893
3. Khasawneh, A. et al. (2017) Myeloid but not plasmacytoid blood DCs possess Th1 polarizing and Th1/Th17 recruiting capacity in psoriasis. Immunol. Lett. 189: 109-113
4. Karyampudi, I. et al. (2016) PD-1 blunts the function of ovarian tumor-infiltrating dendritic cells by inactivating NF-κB. Cancer Res. 76(2): 239-250
5. van Beek, J. J. et al. (2016) Human blood myeloid and plasmacytoid dendritic cells cross activate each other and synergize in inducing NK cell cytotoxicity. Oncoimmunology 5(10): e1227902
6. Su, S. et al. (2017)
  
  Blocking the recruitment of naive CD4
  ⁺
  T cells reverses immunosuppression in breast cancer.
  
  Cell Res. 27(4): 461-482

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BDCA-1 Cell Isolation Kit, human

For research use only

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Background information

Downstream applications

Columns

Figure 1

Selected references

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