The monoclonal antibody DCGM4 recognizes Langerin, a protein specific of Langerhans cells, and is rapidly internalized from the cell surface

• Published in: European journal of immunology Vol. 29; no. 9; pp. 2695 - 2704
• Main Authors: Valladeau, Jenny, Duvert‐Frances, Valérie, Pin, Jean‐Jacques, Dezutter‐Dambuyant, Colette, Vincent, Claude, Massacrier, Catherine, Vincent, Jérôme, Yoneda, Kozo, Banchereau, Jacques, Caux, Christophe, Davoust, Jean, Saeland, Sem
• Format: Journal Article
• Language: English
• Published: Weinheim WILEY‐VCH Verlag GmbH 01-09-1999
• Subjects: Antibodies, Monoclonal - metabolism; Antigen-Antibody Reactions; Antigens, CD; Antigens, Surface - biosynthesis; Antigens, Surface - immunology; Antigens, Surface - isolation & purification; Antigens, Surface - physiology; CD40 Antigens - metabolism; CD40 Ligand; Cell Separation; Down-Regulation - immunology; Humans; Internalization; Langerhans cell; Langerhans Cells - chemistry; Langerhans Cells - metabolism; Langerhans Cells - ultrastructure; Langerin; Lectins, C-Type; Ligands; Mannose-Binding Lectins; Membrane Glycoproteins - biosynthesis; Membrane Glycoproteins - immunology; Membrane Glycoproteins - isolation & purification; Membrane Glycoproteins - metabolism; Membrane Glycoproteins - physiology; MHC class II; Molecular Weight; Monoclonal antibody
• ISSN: 0014-2980; 1521-4141
• DOI: 10.1002/(SICI)1521-4141(199909)29:09<2695::AID-IMMU2695>3.0.CO;2-Q

We generated monoclonal antibody (mAb) DCGM4 by immunization with human dendritic cells (DC) from CD34+ progenitors cultured with granulocyte‐macrophage colony‐stimulating factor and TNF‐α. mAb DCGM4 was selected for its reactivity with a cell surface epitope present only on a subset of DC. Reactivity was strongly enhanced by the Langerhans cell (LC) differentiation factor TGF‐β and down‐regulated by CD40 ligation. mAb DCGM4 selectively stained LC, hence we propose that the antigen be termed Langerin. mAb DCGM4 also stained intracytoplasmically, but neither colocalized with MHC class II nor with lysosomal LAMP‐1 markers. Notably, mAb DCGM4 was rapidly internalized at 37 °C, but did not gain access to MHC class II compartments. Finally, Langerin was immunoprecipitated as a 40‐kDa protein with a pI of 5.2 – 5.5. mAb DCGM4 will be useful to further characterize Langerin, an LC‐restricted molecule involved in routing of cell surface material in immature DC.