title: Novel antibodies directed against the human erythropoietin receptor: creating a basis for clinical implementation.
abstract: Recombinant human erythropoietin (rHuEPO) is an effective treatment for anaemia but concerns that it causes disease progression in cancer patients by activation of EPO receptors (EPOR) in tumour tissue have been controversial and have restricted its clinical use. Initial clinical studies were flawed because they used polyclonal antibodies, later shown to lack specificity for EPOR. Moreover, multiple isoforms of EPOR caused by differential splicing have been reported in cancer cell lines at the mRNA level but investigations of these variants and their potential impact on tumour progression, have been hampered by lack of suitable antibodies. The EpoCan consortium seeks to promote improved pathological testing of EPOR, leading to safer clinical use of rHuEPO, by producing well characterized EPOR antibodies. Using novel genetic and traditional peptide immunization protocols, we have produced mouse and rat monoclonal antibodies, and show that several of these specifically recognize EPOR by Western blot, immunoprecipitation, immunofluorescence, flow cytometry and immunohistochemistry in cell lines and clinical material. Widespread availability of these antibodies should enable the research community to gain a better understanding of the role of EPOR in cancer, and eventually to distinguish patients who can be treated safely by rHuEPO from those at increased risk from treatment.
authors.affiliation: Northern Ireland Molecular Pathology Laboratory, Belfast Health & Social Care Trust, Queen's University Belfast, Belfast, UK; Centre for Cancer Research and Cell Biology, Queen's University Belfast, Belfast, UK.
authors.forename: Perry
authors.initials: P
authors.lastname: Maxwell
authors.forename: Florinda
authors.initials: F
authors.lastname: Melendez-Rodríguez
authors.forename: Kyle B
authors.initials: KB
authors.lastname: Matchett
authors.forename: Julian
authors.initials: J
authors.lastname: Aragones
authors.forename: Nathalie
authors.initials: N
authors.lastname: Ben-Califa
authors.forename: Heidelinde
authors.initials: H
authors.lastname: Jaekel
authors.forename: Ludger
authors.initials: L
authors.lastname: Hengst
authors.forename: Herbert
authors.initials: H
authors.lastname: Lindner
authors.forename: Andre
authors.initials: A
authors.lastname: Bernardini
authors.forename: Ulf
authors.initials: U
authors.lastname: Brockmeier
authors.forename: Joachim
authors.initials: J
authors.lastname: Fandrey
authors.forename: Fritz
authors.initials: F
authors.lastname: Grunert
authors.forename: Howard S
authors.initials: HS
authors.lastname: Oster
authors.forename: Moshe
authors.initials: M
authors.lastname: Mittelman
authors.forename: Mohamed
authors.initials: M
authors.lastname: El-Tanani
authors.forename: Markus
authors.initials: M
authors.lastname: Thiersch
authors.forename: Edith M
authors.initials: EM
authors.lastname: Schneider Gasser
authors.forename: Max
authors.initials: M
authors.lastname: Gassmann
authors.forename: David
authors.initials: D
authors.lastname: Dangoor
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Cuthbert
authors.forename: Alexandra
authors.initials: A
authors.lastname: Irvine
authors.forename: Anne
authors.initials: A
authors.lastname: Jordan
authors.forename: Terence
authors.initials: T
authors.lastname: Lappin
authors.forename: John
authors.initials: J
authors.lastname: Thompson
authors.forename: Drorit
authors.initials: D
authors.lastname: Neumann
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D060326:Chemistry Techniques, Synthetic / Q000379:methods; D005434:Flow Cytometry / Q000379:methods; D005455:Fluorescent Antibody Technique; D020868:Gene Silencing; D006801:Humans; D047468:Immunoprecipitation; D051379:Mice; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D051381:Rats; D017467:Receptors, Erythropoietin / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D018570:Risk Assessment / Q000379:methods; D009626:Terminology as Topic; D014407:Tumor Cells, Cultured / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D009363:Neoplasm Proteins; D017467:Receptors, Erythropoietin
keywords: antibody; cancer anaemia; erythropoietin receptor; recombinant erythropoietin; risk assessment