title: Engagement of two distinct binding domains on CCL17 is required for signaling through CCR4 and establishment of localized inflammatory conditions in the lung.
abstract: CCL17 (TARC) function can be completely abolished by mAbs that block either one of two distinct sites required for CCR4 signaling. This chemokine is elevated in sera of asthma patients and is responsible for establishing inflammatory sites through CCR4-mediated recruitment of immune cells. CCL17 shares the GPCR CCR4, with CCL22 (MDC) but these two chemokines differentially affect the immune response. To better understand chemokine mediated effects through CCR4, we have generated chimeric anti-mouse CCL17 surrogate antibodies that inhibit function of this ligand in vitro and in vivo. The affinities of the surrogate antibodies for CCL17 range from 685 pM for B225 to 4.9 nM for B202. One antibody, B202, also exhibits weak binding to CCL22 (KD∼2 µM) and no binding to CCL22 is detectable with the second antibody, B225. In vitro, both antibodies inhibit CCL17-mediated calcium mobilization, β-arrestin recruitment and chemotaxis; B202 can also partially inhibit CCL22-mediated β-arrestin recruitment. Both B202 and B225 antibodies neutralize CCL17 in vivo as demonstrated by reduction of methacholine-induced airway hyperreactivity in the A. fumigatus model of asthma. That both antibodies block CCL17 function but only B202 shows any inhibition of CCL22 function suggests that they bind CCL17 at different sites. Competition binding studies confirm that these two antibodies recognize unique epitopes that are non-overlapping despite the small size of CCL17. Taking into consideration the data from both the functional and binding studies, we propose that effective engagement of CCR4 by CCL17 involves two distinct binding domains and interaction with both is required for signaling.
authors: Janssen Research & Development, Spring House, Pennsylvania, United States of America ; Department of Pathology, University of Michigan, Ann Arbor, Michigan, United States of America.
authors: Sandra
authors: S
authors: Santulli-Marotto
authors: Ken
authors: K
authors: Boakye
authors: Eilyn
authors: E
authors: Lacy
authors: Sheng-Jiun
authors: SJ
authors: Wu
authors: Jennifer
authors: J
authors: Luongo
authors: Karl
authors: K
authors: Kavalkovich
authors: Ana
authors: A
authors: Coelho
authors: Cory M
authors: CM
authors: Hogaboam
authors: Mary
authors: M
authors: Ryan
mesh_terms: D000818:Animals; D057134:Antibodies, Neutralizing / Q000276:immunology; D054414:Chemokine CCL17 / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism*; D054422:Chemokine CCL22 / Q000378:metabolism; D005260:Female; D008168:Lung / Q000378:metabolism* / Q000473:pathology*; D051379:Mice; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D054398:Receptors, CCR4 / Q000378:metabolism*; D015398:Signal Transduction
chemical_list: D057134:Antibodies, Neutralizing; C499538:Ccl17 protein, mouse; C515355:Ccr4 protein, mouse; D054414:Chemokine CCL17; D054422:Chemokine CCL22; D054398:Receptors, CCR4