Limited Cross-Linking of 4-1BB by 4-1BB Ligand and the Agonist Monoclonal Antibody Utomilumab

Yan Li; Shuguang Tan; Chang Zhang; Yan Chai; Mengnan He; Catherine W-H Zhang; Qihui Wang; Zhou Tong; Kefang Liu; Yifan Lei; William J Liu; Yingxia Liu; Zhigang Tian; Xuetao Cao; Jinghua Yan; Jianxun Qi; Po Tien; Shan Gao; George F Gao

doi:10.1016/j.celrep.2018.09.073

Limited Cross-Linking of 4-1BB by 4-1BB Ligand and the Agonist Monoclonal Antibody Utomilumab

Cell Rep. 2018 Oct 23;25(4):909-920.e4. doi: 10.1016/j.celrep.2018.09.073.

Authors

Yan Li¹, Shuguang Tan², Chang Zhang³, Yan Chai⁴, Mengnan He¹, Catherine W-H Zhang⁵, Qihui Wang⁶, Zhou Tong⁷, Kefang Liu⁸, Yifan Lei⁴, William J Liu⁹, Yingxia Liu¹⁰, Zhigang Tian¹¹, Xuetao Cao¹², Jinghua Yan⁶, Jianxun Qi¹, Po Tien¹³, Shan Gao¹⁴, George F Gao¹⁵

Affiliations

¹ CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China; University of Chinese Academy of Sciences, Beijing 100049, China.
² CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China; Shenzhen Key Laboratory of Pathogen and Immunity, Shenzhen Third People's Hospital, Shenzhen 518112, China.
³ CAS Key Laboratory of Bio-medical Diagnostics, Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou 215163, China.
⁴ CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China.
⁵ ImmuFuCell Biotechnology, Beijing 100102, China.
⁶ Shenzhen Key Laboratory of Pathogen and Immunity, Shenzhen Third People's Hospital, Shenzhen 518112, China; CAS Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China.
⁷ Shenzhen Key Laboratory of Pathogen and Immunity, Shenzhen Third People's Hospital, Shenzhen 518112, China; CAS Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China; Shanxi Academy of Advanced Research and Innovation, Taiyuan 030032, China.
⁸ University of Chinese Academy of Sciences, Beijing 100049, China; National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention (China CDC), Beijing 102206, China.
⁹ Shenzhen Key Laboratory of Pathogen and Immunity, Shenzhen Third People's Hospital, Shenzhen 518112, China; National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention (China CDC), Beijing 102206, China.
¹⁰ Shenzhen Key Laboratory of Pathogen and Immunity, Shenzhen Third People's Hospital, Shenzhen 518112, China.
¹¹ Institute of Immunology, School of Life Sciences and Medical Center, University of Science and Technology of China, Hefei 230027, China.
¹² Department of Immunology & Center for Immunotherapy, Institute of Basic Medical Sciences, Peking Union Medical College, Chinese Academy of Medical Sciences, Beijing 100005, China.
¹³ CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China; University of Chinese Academy of Sciences, Beijing 100049, China. Electronic address: tienpo@im.ac.cn.
¹⁴ CAS Key Laboratory of Bio-medical Diagnostics, Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou 215163, China; Shanxi Academy of Advanced Research and Innovation, Taiyuan 030032, China. Electronic address: gaos@sibet.ac.cn.
¹⁵ CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China; University of Chinese Academy of Sciences, Beijing 100049, China; Shenzhen Key Laboratory of Pathogen and Immunity, Shenzhen Third People's Hospital, Shenzhen 518112, China; Shanxi Academy of Advanced Research and Innovation, Taiyuan 030032, China; National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention (China CDC), Beijing 102206, China. Electronic address: gaof@im.ac.cn.

PMID: 30355497
DOI: 10.1016/j.celrep.2018.09.073

Abstract

Monoclonal antibodies (mAbs) targeting the co-stimulatory molecule 4-1BB are of interest for tumor immunotherapy. We determined the complex structures of human 4-1BB with 4-1BB ligand (4-1BBL) or utomilumab to elucidate the structural basis of 4-1BB activation. The 4-1BB/4-1BBL complex displays a typical TNF/TNFR family binding mode. The structure of utomilumab/4-1BB complex shows that utomilumab binds to dimeric 4-1BB with a distinct but partially overlapping binding area with 4-1BBL. Competitive binding analysis demonstrates that utomilumab blocks the 4-1BB/4-1BBL interaction, indicating the interruption of ligand-mediated signaling. The binding profiles of 4-1BBL and utomilumab to monomeric or dimeric 4-1BB indicate limited cross-linking of 4-1BB molecules. These findings provide mechanistic insight into the binding of 4-1BB with its ligand and its agonist mAb, which may facilitate the future development of anti-4-1BB biologics for tumor immunotherapy.

Keywords: 4-1BB; 4-1BB ligand; 4-1BBL; agonist antibody; complex structure; cross-linking; utomilumab.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

4-1BB Ligand / chemistry
4-1BB Ligand / metabolism*
Adult
Amino Acid Sequence
Antibodies, Monoclonal / metabolism*
Antibodies, Monoclonal, Humanized
Binding, Competitive
Cross-Linking Reagents / metabolism*
Female
Humans
Immunoglobulin G / metabolism*
Models, Molecular
Protein Binding
Protein Multimerization
Protein Structure, Secondary
Single-Chain Antibodies / chemistry
Single-Chain Antibodies / metabolism
Tumor Necrosis Factor Receptor Superfamily, Member 9 / agonists*
Tumor Necrosis Factor Receptor Superfamily, Member 9 / chemistry
Tumor Necrosis Factor Receptor Superfamily, Member 9 / metabolism

Substances

4-1BB Ligand
Antibodies, Monoclonal
Antibodies, Monoclonal, Humanized
Cross-Linking Reagents
Immunoglobulin G
Single-Chain Antibodies
Tumor Necrosis Factor Receptor Superfamily, Member 9
utomilumab