title: Expression analysis and specific blockade of the receptor for human thymic stromal lymphopoietin (TSLP) by novel antibodies to the human TSLPRα receptor chain.
abstract: Thymic stromal lymphopoietin (TSLP) is an interleukin-7 (IL-7)-like cytokine with a pivotal role in development and maintenance of atopic diseases such as allergic asthma and atopic dermatitis. Moreover, recent studies show an involvement of TSLP in the progression of various cancers. TSLP signaling is mediated by the TSLP receptor (TSLPR), a heterodimeric type I cytokine receptor. It consists of the IL-7 receptor alpha chain (IL-7Rα), which is shared with the IL-7 receptor, and the TSLPRα chain as a specific subunit. Blocking signal release by TSLP without affecting IL-7 function is a potentially interesting option for the treatment of atopic diseases or certain tumors. By employing the extracellular domain of human TSLPRα chain (hTSLPRα(ex)) as an antigen, we generated a set of monoclonal antibodies. Several binders to native and/or denatured receptor protein were identified and characterized by cytometry and Western blot analysis. A screen based on a STAT3-driven reporter gene assay in murine pro-B cells expressing a functional hTSLPR yielded two hybridoma clones with specific antagonistic properties towards hTSLP, but not IL-7. Kinetic studies measuring blockade of hTSLP-dependent STAT phosphorylation in a TSLP-responsive cell line revealed an inhibitory constant in the nanomolar range.
authors.affiliation: Institute of Biochemistry II, Jena University Hospital, Jena, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Borowski
authors.forename: Tina
authors.initials: T
authors.lastname: Vetter
authors.forename: Michael
authors.initials: M
authors.lastname: Kuepper
authors.forename: Andreas
authors.initials: A
authors.lastname: Wohlmann
authors.forename: Sebastian
authors.initials: S
authors.lastname: Krause
authors.forename: Thomas
authors.initials: T
authors.lastname: Lorenzen
authors.forename: Johann Christian
authors.initials: JC
authors.lastname: Virchow
authors.forename: Werner
authors.initials: W
authors.lastname: Luttmann
authors.forename: Karlheinz
authors.initials: K
authors.lastname: Friedrich
mesh_terms: D000818:Animals; D019138:Antibodies, Blocking / Q000494:pharmacology*; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D000941:Antigens / Q000378:metabolism; D005434:Flow Cytometry; D057809:HEK293 Cells; D006801:Humans; D008024:Ligands; D051379:Mice; D008807:Mice, Inbred BALB C; D011485:Protein Binding / Q000187:drug effects; D011489:Protein Denaturation / Q000187:drug effects; D018121:Receptors, Cytokine / Q000037:antagonists & inhibitors* / Q000097:blood / Q000737:chemistry / Q000276:immunology*; D011994:Recombinant Proteins / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D012995:Solubility
chemical_list: D019138:Antibodies, Blocking; D000911:Antibodies, Monoclonal; D000941:Antigens; C498762:CRLF2 protein, human; D008024:Ligands; D018121:Receptors, Cytokine; D011994:Recombinant Proteins