|Detection of CLEC9a in HEK293 Human Cell Line Transfected with Human CLEC9a and eGFP by Flow Cytometry. HEK293 human embryonic kidney cell line transfected with human CLEC9a and eGFP was stained with either (A) Mouse Anti-Human CLEC9a APC-conjugated Monoclonal Antibody (Catalog # FAB6049A) or (B) Mouse IgG1 Allophycocyanin Isotype Control (Catalog # IC002A). View our protocol for Staining Membrane-associated Proteins.|
|Detection of CLEC9a in Human Peripheral Blood Cells by Flow Cytometry. Human peripheral blood cells gated on CD3-CD141+ cells were stained with Mouse Anti-Human HLA-DR PE-conjugated Monoclonal Antibody (Catalog # FAB4869P) and either (A) Mouse Anti-Human CLEC9a APC-conjugated Monoclonal Antibody (Catalog # FAB6049A) or (B) Mouse IgG1 Allophycocyanin Isotype Control (Catalog # IC002A). View our protocol for Staining Membrane-associated Proteins.|
CLEC9a (C-type lectin domain family 9 member A), also known as DNGR-1, is a type II transmembrane glycoprotein member of the C‑type lectin superfamily (1, 2). Mature human CLEC9a consists of a 35 amino acid (aa) cytoplasmic domain with an ITAM-like motif, a 21 aa transmembrane segment, and a 185 extracellular domain (ECD) that contains a stalk region and one C-type lectin domain (CTLD) (3‑5). Within the ECD, human CLEC9a shares 57% aa sequence identity with mouse and rat CLEC9a. Although the CTLD of CLEC9a structurally resembles that of other C-type lectins, it lacks the conserved residues that typically mediate calcium and carbohydrate binding. CLEC9a is expressed as a disulfide-linked homodimer of approximately 50 kDa N-glycosylated subunits (3, 5). Human CLEC9a expression is restricted to a subpopulation of BDCA-3+ conventional dendritic cells (cDC) and CD16- monocytes (3‑5). BDCA-3+ cDC are analagous to mouse CD8+ cDC which are specialized in antigenic cross-presentation in complex with MHC class I molecules (6). In mouse, CLEC9a is additionally expressed on plasmacytoid dendritic cells (4, 5). CLEC9a ligation enhances antigen uptake and processing, leading to presentation on MHC class I and cytotoxic T cell (CTL) priming (3‑5). In mouse, CLEC9a recognizes normally intracellular determinant(s) of necrotic cells and mediates their uptake by the dendritic cell (7). The subsequent antigenic cross-presentation to CTL is important for clearing necrotic cellular debris (7). CLEC9a signaling triggers activation of the tyrosine kinase Syk (3, 7).
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