title: Surrogate antibodies that specifically bind and neutralize CCL17 but not CCL22.
abstract: The chemokines CCL17 (TARC) and CCL22 (MDC) function through the same receptor, CCR4, but have been proposed to differentially affect the immune response. To better understand the role of the individual ligands, a panel of rat anti-mouse CCL17 surrogate antibodies was generated that can be used to differentiate CCL17 and CCL22 function in vitro and in vivo. We have successfully identified a panel of neutralizing antibodies by screening hybridomas for the ability to inhibit CCL17-mediated calcium mobilization. Chemotaxis in response to CCL17 is also inhibited, providing further evidence that the antibodies in this panel are antagonistic. Using a recombinant cell line expressing human CCR4, we show that the antibodies block β-arrestin recruitment as evidence that the antibodies are specifically blocking CCL17 signaling through CCR4. The antibodies within this panel inhibit calcium mobilization with varying potency in the calcium flux assay, having apparent IC50 ranging from approximately 1 to >400 ng/mL. Although both CCL17 and CCL22 function through CCR4, only a single antibody was identified as having detectable binding to CCL22. This panel of CCL17-specific antibodies provides tools that can be used to differentiate CCL17 and CCL22 function through CCR4 interaction in vitro and in vivo.
authors.affiliation: Janssen Research & Development, Radnor, Pennsylvania 19087, USA. ssantull@its.jnj.com
authors.forename: Sandra
authors.initials: S
authors.lastname: Santulli-Marotto
authors.forename: Jamie
authors.initials: J
authors.lastname: Fisher
authors.forename: Theodore
authors.initials: T
authors.lastname: Petley
authors.forename: Ken
authors.initials: K
authors.lastname: Boakye
authors.forename: Tadas
authors.initials: T
authors.lastname: Panavas
authors.forename: Jennifer
authors.initials: J
authors.lastname: Luongo
authors.forename: Karl
authors.initials: K
authors.lastname: Kavalkovich
authors.forename: Michael
authors.initials: M
authors.lastname: Rycyzyn
authors.forename: Bingyuan
authors.initials: B
authors.lastname: Wu
authors.forename: Lester
authors.initials: L
authors.lastname: Gutshall
authors.forename: Ana
authors.initials: A
authors.lastname: Coelho
authors.forename: Cory M
authors.initials: CM
authors.lastname: Hogaboam
authors.forename: Mary
authors.initials: M
authors.lastname: Ryan
mesh_terms: D000818:Animals; D057134:Antibodies, Neutralizing / Q000276:immunology*; D000915:Antibody Affinity / Q000276:immunology; D019390:Arrestins / Q000276:immunology; D002460:Cell Line; D054414:Chemokine CCL17 / Q000276:immunology*; D054422:Chemokine CCL22 / Q000276:immunology*; D002633:Chemotaxis / Q000276:immunology; D006801:Humans; D051381:Rats; D054398:Receptors, CCR4 / Q000276:immunology*; D000071557:beta-Arrestins
chemical_list: D057134:Antibodies, Neutralizing; D019390:Arrestins; C515354:CCR4 protein, human; D054414:Chemokine CCL17; D054422:Chemokine CCL22; D054398:Receptors, CCR4; D000071557:beta-Arrestins