Blocking monoclonal antibodies specific for mouse IFN-alpha/beta receptor subunit 1 (IFNAR-1) from mice immunized by in vivo hydrodynamic transfection.
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.
Sheehan
Kathleen C F
KC
Department of Pathology and Immunology, Center for Immunology, Washington University School of Medicine, St. Louis, MO 63110, USA.
Lai
Koon Siew
KS
Dunn
Gavin P
GP
Bruce
Allen T
AT
Diamond
Mark S
MS
Heutel
Jennifer D
JD
Dungo-Arthur
Corazon
C
Carrero
Javier A
JA
White
J Michael
JM
Hertzog
Paul J
PJ
Schreiber
Robert D
RD
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
D019138:Antibodies, Blocking; D000911:Antibodies, Monoclonal; C508032:Ifnar1 protein, mouse; D053633:Receptor, Interferon alpha-beta; D053634:TYK2 Kinase; C499356:Tyk2 protein, mouse