|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 PerCP‑conjugated Monoclonal Antibody (Catalog # FAB4869C) and either (A) Mouse Anti-Human CLEC9a Alexa Fluor® 488‑conjugated Monoclonal Antibody (Catalog # FAB6049G) or (B) Mouse IgG1 Alexa Fluor 488 Isotype Control (Catalog # IC002G). 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).