title: Identification of the Single Immunodominant Region of the Native Human CC Chemokine Receptor 6 Recognized by Mouse Monoclonal Antibodies.
abstract: Chemokines and their receptors play an important role in cell trafficking and recruitment. The CCR6 chemokine receptor, selectively expressed on leukocyte populations, has been shown to play a deleterious role in the pathogenesis of various chronic inflammatory diseases and, as such, may constitute a prime target in the development of immunotherapeutic treatment. However, to date no neutralizing mouse monoclonal antibodies (mAbs) specific for this chemokine receptor have been reported, whereas information on small molecules capable of interfering with the interaction of CCR6 and its ligands is scant. Here, we report the failure to generate neutralizing mouse mAbs specific for human (hu)CCR6. Immunization of mice with peptides mimicking extracellular domains, potentially involved in CCR6 function, failed to induce Abs reactive with the native receptor. Although the use of NIH-3T3 cells expressing huCCR6 resulted in the isolation of mAbs specific for this receptor, they were not able to block the interaction between huCCR6 and huCCL20. Investigation of the anti-huCCR6 mAbs generated in the present study, as well as those commercially available, show that all mAbs invariably recognize a unique, non-neutralizing, immunodominant region in the first part of its N-terminal domain. Together, these results indicate that the generation of potential neutralizing anti-huCCR6 mAbs in the mouse is unlikely to succeed and that alternative techniques, such as the use of other animal species for immunization, might constitute a better approach to generate such a potentially therapeutic tool for the treatment of inflammatory disease.
authors.affiliation: Sorbonne Universités, Inserm U1135, UPMC Univ Paris 06 UMRS 1135, Centre d'Immunologie et des Maladies Infectieuses (Cimi-Paris), F-75013, Paris, France.
authors.forename: Karim
authors.initials: K
authors.lastname: Dorgham
authors.affiliation: Institut de Recherche en Cancérologie, F-34090, Montpellier, France.
authors.forename: Cécile
authors.initials: C
authors.lastname: Dejou
authors.affiliation: Sorbonne Universités, UPMC Univ Paris 06, IBPS-FR 3631, Service de Synthèse Peptidique, F-75005, Paris, France.
authors.forename: Christophe
authors.initials: C
authors.lastname: Piesse
authors.affiliation: Sorbonne Universités, Inserm U1135, UPMC Univ Paris 06 UMRS 1135, Centre d'Immunologie et des Maladies Infectieuses (Cimi-Paris), F-75013, Paris, France.|AP-HP, Groupement Hospitalier Pitié-Salpêtrière, Département d'Immunologie, F-75013, Paris, France.
authors.forename: Guy
authors.initials: G
authors.lastname: Gorochov
authors.affiliation: Institute for Regenerative Medicine and Biotherapy, Inserm U1183, F-34295, Montpellier, France.
authors.forename: Jérôme
authors.initials: J
authors.lastname: Pène
authors.affiliation: Sorbonne Universités, Inserm U1135, UPMC Univ Paris 06 UMRS 1135, Centre d'Immunologie et des Maladies Infectieuses (Cimi-Paris), F-75013, Paris, France.
authors.forename: Hans
authors.identifier: http://orcid.org/0000-0001-7454-1836
authors.initials: H
authors.lastname: Yssel
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D057134:Antibodies, Neutralizing / Q000276:immunology*; D001666:Binding Sites, Antibody; D002460:Cell Line; D054418:Chemokine CCL20 / Q000276:immunology; D005260:Female; D006801:Humans; D007114:Immunization; D016056:Immunodominant Epitopes / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D041681:NIH 3T3 Cells; D054399:Receptors, CCR6 / Q000008:administration & dosage / Q000276:immunology*; D011994:Recombinant Proteins / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D057134:Antibodies, Neutralizing; C108355:CCL20 protein, human; C106311:CCR6 protein, human; D054418:Chemokine CCL20; D016056:Immunodominant Epitopes; D054399:Receptors, CCR6; D011994:Recombinant Proteins