title: Generation and characterization of rat anti-mouse ST2L monoclonal antibodies.
abstract: ST2L is a transmembrane receptor that belongs to the IL-1 receptor family. The receptor is expressed on various cell types including Th2 cells, mast cells, basophils, growth-activated fibroblasts, and vascular endothelial cells. ST2L activation by its ligand IL-33 has been implicated in Th2-mediated immunity, inflammation, and allergic responses in vivo. Inhibition of ST2L activity can attenuate Th2-dominated immune responses such as lung eosinophilia, airway hyper-responsiveness, and arthritis in animal models. Here we report the generation and in vitro characterization of a panel of rat anti-mouse ST2L monoclonal antibodies. We demonstrate that the antibodies specifically bind to recombinant receptor protein and that a subset of the binders inhibits mouse ST2L activity in multiple in vitro assays. Four of the identified anti-mouse ST2L antibodies were shown to prevent IL-33 from binding to ST2L, down-regulate IL-33-induced NF-κB signaling, and neutralize the ability of IL-33 to stimulate mouse Th2 cell proliferation. The characterized monoclonal antibodies are important tools that will be used to study mouse ST2L receptor functionality in vivo.
authors.affiliation: Biologics Research, Centocor, Radnor, Pennsylvania, USA. nfursov@its.jnj.com
authors.forename: Natalie
authors.initials: N
authors.lastname: Fursov
authors.forename: Erin
authors.initials: E
authors.lastname: Johnston
authors.forename: Karen
authors.initials: K
authors.lastname: Duffy
authors.forename: Adam
authors.initials: A
authors.lastname: Cotty
authors.forename: Theodore
authors.initials: T
authors.lastname: Petley
authors.forename: Jamie
authors.initials: J
authors.lastname: Fisher
authors.forename: Haiyan
authors.initials: H
authors.lastname: Jiang
authors.forename: Michael A
authors.initials: MA
authors.lastname: Rycyzyn
authors.forename: Jill
authors.initials: J
authors.lastname: Giles-Komar
authors.forename: Gordon
authors.initials: G
authors.lastname: Powers
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology / Q000302:isolation & purification / Q000494:pharmacology; D000918:Antibody Specificity; D001710:Biotin / Q000737:chemistry / Q000378:metabolism; D049109:Cell Proliferation / Q000187:drug effects; D015536:Down-Regulation; D057809:HEK293 Cells; D006801:Humans; D006825:Hybridomas / Q000276:immunology / Q000378:metabolism; D006967:Hypersensitivity / Q000188:drug therapy / Q000276:immunology; D018796:Immunoconjugates / Q000737:chemistry / Q000378:metabolism; D007249:Inflammation / Q000188:drug therapy / Q000276:immunology; D007375:Interleukin-1 / Q000276:immunology; D000067596:Interleukin-33; D007378:Interleukins / Q000276:immunology* / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D016328:NF-kappa B / Q000378:metabolism; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017472:Receptors, Interleukin-1 / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects* / Q000276:immunology; D018418:Th2 Cells / Q000187:drug effects* / Q000276:immunology; D014162:Transfection
chemical_list: D000911:Antibodies, Monoclonal; C506535:Il33 protein, mouse; D018796:Immunoconjugates; D007375:Interleukin-1; D000067596:Interleukin-33; D007378:Interleukins; D016328:NF-kappa B; D017472:Receptors, Interleukin-1; D011994:Recombinant Proteins; C530318:ST2L protein, mouse; D001710:Biotin