Identification of a novel, dendritic cell-associated molecule, dectin-1, by subtractive cDNA cloning.

摘要: Dendritic cells (DC) are special subsets of antigen presenting characterized by their potent capacity to activate immunologically naive T cells. By subtracting the mRNAs expressed mouse epidermus-derived DC line XS52 with J774 macrophage line, we identified five novel genes that were selectively this line. One these encoded a type II membrane-integrated polypeptide 244 amino acids containing putative carbohydrate recognition domain motif at COOH-terminal end. This molecule, termed "dectin-1," was abundantly both mRNA and protein levels but not non-DC lines (including line). Dectin-1 detected predominantly in spleen thymus (by Northern blotting) skin-resident DC, i.e. Langerhans reverse transcription-polymerase chain reaction). Affinity-purified antibody against dectin-1 43-kDa glycoprotein membrane fractions isolated from cDNA-transfected COS-1 His-tagged recombinant proteins extracellular domains showed marked specific binding surface promoted proliferation presence anti-CD3 monoclonal suboptimal concentrations. These vitro results suggest on may bind as yet undefined ligand(s) cells, thereby delivering cell co-stimulatory signals. Not only do document efficacy subtractive cDNA cloning for identification unique they also provide framework studying physiological function dectin-1.