CD303 (BDCA-2)

Unique antibody for DC research
Get accurate PDC counts in whole blood, bone marrow, and tissue sections

Overview

The antibody is suitable for the direct identification of CD303 (BDCA-2)⁺ plasmacytoid dendritic cells in blood, lymphoid and non-lymphoid tissue by immunofluorescent (or immunocytochemical) staining and flow cytometric analysis.^{1,3, 5–11} It can be use for staining of fresh or formaldehyde-fixed cells in suspension and for staining of, for example, air-dried, acetone-fixed cryosections.^3,5 Staining with CD303 (BDCA-2) antibodies is recommended for flow cytometric evaluation if plasmacytoid dendritic cells are isolated using the CD304 CD304 (BDCA-4/Neuropilin-1) MicroBead Kit.³

Details

Background information
CD303 (BDCA-2) antibodies have been used, for example, to identify, characterize, and enumerate plasmacytoid dendritic cells in whole blood of healthy and HIV-infected individuals, and for analyzing the role of DC-SIGN in HIV infection and transmission.^7,11 Furthermore, CD303 (BDCA-2) antibodies were used to identify and enumerate plasmacytoid dendritic cells in blood and bone marrow samples before and after hematopoietic stem cell mobilization⁸ or transplantation¹⁰. CD303 (BDCA-2) antibodies were also used for immunohistochemical staining, for example to identify plasmacytoid dendritic cells in tissue sections from patients with different inflammatory skin diseases.^5,12,13
Clone AC144 recognizes the the CD303 (BDCA-2) antigen^1,3 which is expressed on human plasmacytoid dendritic cells in blood, lymphoid (e.g. tonsils and bone marrow) and non-lymphoid tissue.^3,6 Specific expression allows direct identification of plasmacytoid dendritic cells using just one marker.^1–11 CD303 (BDCA-2)⁺ plasmacytoid dendritic cells in blood and bone marrow are CD11c^–, CD123^high, CD4⁺, Lin^–, CD45RA⁺, CD304 (BDCA-4/Neuropilin-1)⁺, CD141 (BDCA-3)^low, CD1c (BDCA-1)^–, CD14^–, and CD2^–. They express neither myeloid lineage markers (CD13, CD33) nor Fc receptors (CD32, CD64, FcRI).^1,6 CD303 (BDCA-2) is strongly expressed on freshly isolated plasmacytoid dendritic cells but down-regulated within 48 hours of culturing.¹ Unlike CD304 (BDCA-4/Neuropilin-1), CD303 (BDCA-2) is not detectable on ex vivo generated monocyte-derived or hematopoietic precursor cell-derived CD1a⁺ dendritic cells.^1,9
The CD303 (BDCA-2) antigen is a novel type II transmembrane C-type lectin.³ Remarkably, plasmacytoid dendritic cells can take up ligands via CD303 (BDCA-2), then process and present the ligands to T cells.^1,3 Unlike binding of antibodies to CD304 (BDCA-4), binding of antibodies to CD303 (BDCA-2) inhibits type I IFN production, which is induced in plasmacytoid dendritic cells by, for example, the influenza virus.^3,6

Cross-reactivity

Clone AC144 has been tested to cross-react with

Cynomolgus monkey (Macaca fascicularis)

It does not react with cells from rhesus macaque (Macaca mulatta).

Clone	Isotype
AC144	Mouse IgG1

Figure 1

Human PBMCs were stained with CD303 (BDCA-2), lineage marker (CD3, CD16, CD14, CD19, CD20, CD56), CD123, and CD11c antibodies. Flow cytometric analysis confirms that CD303 (BDCA-2)⁺ plasmacytoid dendritic cells are Lin^–, CD123⁺ and CD11c^–, and that CD303 (BDCA-2) is exclusively expressed on plasmacytoid dendritic cells.

Figure 2

Direct identification of plasmacytoid dendritic cells in cryosections of tonsils by immunofluorescent staining. Sections were stained with CD303 (BDCA-2)-FITC (green) and non-conjugated monoclonal antibody against CD123 (A), CD20 (B), CD8 (C) or HLA-DR (D), respectively. Staining with non-conjugated monoclonal antibodies was revealed by secondary staining with biotinylated anti-isotype antibodies and Texas red-conjugated streptavidin (red).

Figure 3

PBMCs from cynomolgus monkey (Macaca fascicularis) were stained with CD303 (BDCA-2)-APC and Anti-HLA-DR-FITC and analyzed by flow cytometry.

PBMCs from cynomolgus monkey (<i>Macaca fascicularis</i>) were stained with CD303 (BDCA-2)-APC and Anti-HLA-DR-FITC and analyzed by flow cytometry.