Selection of a picomolar antibody that targets CXCR2-mediated neutrophil activation and alleviates EAE symptoms

• Published in: Nature communications 2021-05, Vol.12 (1), p.2547-2547, Article 2547
• Main Authors: Shi, Xiaojie, Wan, Yue, Wang, Nan, Xiang, Jiangchao, Wang, Tao, Yang, Xiaofeng, Wang, Ju, Dong, Xuxue, Dong, Liang, Yan, Lei, Li, Yu, Liu, Lili, Hou, Shinchen, Zhong, Zhenwei, Wilson, Ian A., Yang, Bei, Yang, Guang, Lerner, Richard A.
• Format: Article
• Language: English
• Subjects: 101/1; 13/1; 13/31; 13/95; 14/19; 38/77; 49/1; 64/110; 82/1; 82/51; 96/21; 96/33; 96/95; Antibodies; Antibody libraries; Antigens; Asthma; Binding; Cell activation; Chemical compounds; Chemokine receptors; Chemokines; Chemotaxis; Chronic obstructive pulmonary disease; Combinatorial analysis; Crystal structure; CXC chemokines; CXCR2 protein; Deuterium; Drug development; Epitopes; Experimental allergic encephalomyelitis; G protein-coupled receptors; Glomerulonephritis; Humanities and Social Sciences; Inflammatory bowel diseases; Interleukin 8; Intestine; Leukocyte migration; Leukocytes (neutrophilic); Ligands; Lung cancer; Lung diseases; Mass spectrometry; Mass spectroscopy; multidisciplinary; Neutrophils; Obstructive lung disease; Panning; Pharmacology; Receptors; Science; Science (multidisciplinary); Signs and symptoms; Therapeutic targets; Tumor necrosis factor
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-021-22810-z

Receptors and their ligands are important therapeutic targets for about one third of marketed drugs. Here, we describe an epitope-guided approach for selection of antibodies that modulate cellular signaling of targeted receptors. We chose CXC chemokine receptor 2 (CXCR2) in the G-protein coupled receptor superfamily as receptor and a CXCR2 N-terminal peptide for antibody selection. We obtain a highly selective, tight-binding antibody from a 10 11 -member antibody library using combinatorial enrichment. Structural and Hydrogen-Deuterium-Exchange mass spectrometry analyses demonstrate antibody interaction with an N-terminal region of CXCR2 that is part of the IL-8 epitope. The antibody strongly inhibits IL-8-induced and CXCR2-mediated neutrophil chemotaxis in vitro and alleviates h CXCR2-dependent experimental autoimmune encephalomyelitis symptoms in mice. As inappropriate neutrophil migration accompanies many diseases including inflammatory bowel disease, glomerulonephritis, allergic asthma, chronic obstructive pulmonary disease, and cancer, this antibody has potential for development as a therapeutic agent, akin to anti-TNF antibodies. However, an important difference here is that the antibody targets the chemokine receptor and competes with natural ligand, rather than targeting the ligand itself. CXCR2 is central to neutrophil chemotaxis and hence to some inflammatory diseases. Here the authors demonstrate the value of an epitope-guided antibody panning method to develop a tight binding anti-hCXCR2 antibody, along with crystal structures of this antibody and antigen, that can block neutrophil chemotaxis and protect mice in an EAE model.