title: Generation of anti-DC-SIGN monoclonal antibodies capable of blocking HIV-1 gp120 binding and reactive on formalin-fixed tissue.
abstract: DC-SIGN is a C-type lectin of recognized importance in immunology and in the pathogenicity human pathogens. Monoclonal antibodies directed against DC-SIGN have been generated, but their systemic characterization for interfering with binding of the HIV-1 glycoprotein 120 has often been omitted. Moreover, so far, no anti-DC-SIGN monoclonal antibody has been described that recognizes its antigen after formalin fixation and paraffin embedding. In this study, we have generated new anti-DC-SIGN monoclonal antibodies using HeLa cells stably expressing DC-SIGN as immunogen. We have obtained 11 hybridoma clones producing antibodies that recognized DC-SIGN on monocyte-derived dendritic cells and on dermal-type macrophages. Seven monoclonal antibodies displayed a capacity to interfere with DC-SIGN binding to HIV-1 gp120. One recognized DC-SIGN on formalin-fixed dendritic cells and macrophages. Using this antibody we have obtained specific labelling of DC-SIGN and colocalisation with the dermal macrophage marker CD163 on human skin. The described monoclonal anti-human DC-SIGN antibodies will be of use to the scientific community to address fundamental immunology issues, in particular concerning macrophages and dendritic cells, and help elucidate infection events of pathogen targeting DC-SIGN as recognition receptor.
authors.affiliation: Groupe Immunité des Muqueuses et Agents Pathogènes (G.I.M.A.P.) Université Jean Monnet, Faculté de Médecine, 15 rue Ambroise Paré, 42023 Saint Etienne, France.
authors.forename: Bertrand
authors.initials: B
authors.lastname: Canard
authors.forename: Hortense
authors.initials: H
authors.lastname: Vachon
authors.forename: Thomas
authors.initials: T
authors.lastname: Fontaine
authors.forename: Jean-Jacques
authors.initials: JJ
authors.lastname: Pin
authors.forename: Stéphane
authors.initials: S
authors.lastname: Paul
authors.forename: Christian
authors.initials: C
authors.lastname: Genin
authors.forename: Christopher G
authors.initials: CG
authors.lastname: Mueller
mesh_terms: D000818:Animals; D019138:Antibodies, Blocking / Q000235:genetics / Q000276:immunology*; D058846:Antibodies, Monoclonal, Murine-Derived / Q000235:genetics / Q000276:immunology*; D015703:Antigens, CD / Q000276:immunology; D015214:Antigens, Differentiation, Myelomonocytic / Q000276:immunology; D015815:Cell Adhesion Molecules / Q000235:genetics / Q000276:immunology*; D003713:Dendritic Cells / Q000276:immunology; D020405:Dermis / Q000276:immunology; D005404:Fixatives / Q000737:chemistry; D005557:Formaldehyde / Q000737:chemistry; D015699:HIV Envelope Protein gp120 / Q000235:genetics / Q000276:immunology*; D015497:HIV-1 / Q000276:immunology*; D006367:HeLa Cells; D006801:Humans; D037181:Lectins, C-Type / Q000235:genetics / Q000276:immunology*; D008264:Macrophages / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D009000:Monocytes / Q000276:immunology; D041681:NIH 3T3 Cells; D011956:Receptors, Cell Surface / Q000235:genetics / Q000276:immunology*; D000098902:CD163 Antigen; D000099215:DC-Specific ICAM-3 Grabbing Nonintegrin
chemical_list: D019138:Antibodies, Blocking; D058846:Antibodies, Monoclonal, Murine-Derived; D015703:Antigens, CD; D015214:Antigens, Differentiation, Myelomonocytic; D015815:Cell Adhesion Molecules; D005404:Fixatives; D005557:Formaldehyde; D015699:HIV Envelope Protein gp120; D037181:Lectins, C-Type; D011956:Receptors, Cell Surface; D000098902:CD163 Antigen; D000099215:DC-Specific ICAM-3 Grabbing Nonintegrin; C514793:gp120 protein, Human immunodeficiency virus 1