title: Blocking monoclonal antibodies specific for mouse IFN-alpha/beta receptor subunit 1 (IFNAR-1) from mice immunized by in vivo hydrodynamic transfection.
abstract: Herein we report the generation of mouse monoclonal antibodies (mAbs) specific for the IFNAR-1 subunit of the mouse interferon-alpha/beta (IFN-alpha/beta) receptor (MAR1 mAbs) that block type I IFN receptor signaling and biologic response induction in vitro and in vivo. These mAbs were generated from Ifnar1 (/) mice immunized by in vivo hydrodynamic transfection with a plasmid encoding the extracellular domain (ECD) of murine IFNAR-1. All MAR1 mAbs bound native receptor expressed on cell surfaces and immunoprecipitated IFNAR-1 from solubilized cells, and two mAbs also detected IFNAR-1 by Western blot analysis. in vitro, the mAbs prevented ligand-induced intracellular signaling and induction of a variety of type I IFN-induced biologic responses but had no effect on IFN-gamma-induced responses. The most effective in vitro blocker, MAR1-5A3, also blocked type I IFN-induced antiviral, antimicrobial, and antitumor responses in vivo. We also explored whether murine IFNAR-1 surface expression required the presence of Tyk2. In contrast to Tyk2-deficient human cell lines, comparable IFNAR-1 expression was found on primary cells derived either from wild-type or Tyk2 (/) mice. These mAbs represent much needed tools to more clearly elucidate the biochemistry, cell biology, and physiologic function of the type I IFNs and their receptor in mediating host-protective immunity and immunopathology.
authors.affiliation: Department of Pathology and Immunology, Center for Immunology, Washington University School of Medicine, St. Louis, MO 63110, USA.
authors.forename: Kathleen C F
authors.initials: KC
authors.lastname: Sheehan
authors.forename: Koon Siew
authors.initials: KS
authors.lastname: Lai
authors.forename: Gavin P
authors.initials: GP
authors.lastname: Dunn
authors.forename: Allen T
authors.initials: AT
authors.lastname: Bruce
authors.forename: Mark S
authors.initials: MS
authors.lastname: Diamond
authors.forename: Jennifer D
authors.initials: JD
authors.lastname: Heutel
authors.forename: Corazon
authors.initials: C
authors.lastname: Dungo-Arthur
authors.forename: Javier A
authors.initials: JA
authors.lastname: Carrero
authors.forename: J Michael
authors.initials: JM
authors.lastname: White
authors.forename: Paul J
authors.initials: PJ
authors.lastname: Hertzog
authors.forename: Robert D
authors.initials: RD
authors.lastname: Schreiber
mesh_terms: D000818:Animals; D019138:Antibodies, Blocking / Q000276:immunology* / Q000302:isolation & purification; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000302:isolation & purification; D000918:Antibody Specificity; D015870:Gene Expression; D007114:Immunization / Q000379:methods; D051379:Mice; D018345:Mice, Knockout; D010957:Plasmids / Q000235:genetics / Q000276:immunology; D053633:Receptor, Interferon alpha-beta / Q000037:antagonists & inhibitors / Q000172:deficiency / Q000235:genetics / Q000276:immunology*; D053634:TYK2 Kinase / Q000172:deficiency / Q000276:immunology; D014162:Transfection / Q000379:methods
chemical_list: D019138:Antibodies, Blocking; D000911:Antibodies, Monoclonal; C508032:Ifnar1 protein, mouse; D053633:Receptor, Interferon alpha-beta; D053634:TYK2 Kinase; C499356:Tyk2 protein, mouse