@article{120bd907977445a7a26d6b484a488bd9,
title = "Structure-based Development of Human Interleukin-1β-Specific Antibody That Simultaneously Inhibits Binding to Both IL-1RI and IL-1RAcP",
abstract = "Interleukin-1β (IL-1β) is a potent pleiotropic cytokine playing a central role in protecting cells from microbial pathogen infection or endogenous stress. After it binds to IL-1RI and recruits IL-1 receptor accessory protein (IL-1RAcP), signaling culminates in activation of NF-κB. Many pathophysiological diseases have been attributed to the derailment of IL-1β regulation. Several blocking reagents have been developed based on two mechanisms: blocking the binding of IL-1β to IL-1RI or inhibiting the recruitment of IL-1RAcP to the IL-1β initial complex. In order to simultaneously fulfill these two actions, a human anti-IL-1β neutralizing antibody IgG26 was screened from human genetic phage-display library and furthered structure-optimized to final version, IgG26AW. IgG26AW has a sub-nanomolar binding affinity for human IL-1β. We validated IgG26AW-neutralizing antibodies specific for IL-1β in vivo to prevent human IL-1β-driving IL-6 elevation in C56BL/6 mice. Mice underwent treatments with IgG26AW in A549 and MDA-MB-231 xenograft mouse cancer models have also been observed with tumor shrank and inhibition of tumor metastasis. The region where IgG26 binds to IL-1β also overlaps with the position where IL-1RI and IL-1RAcP bind, as revealed by the 26-Fab/IL-1β complex structure. Meanwhile, SPR experiments showed that IL-1β bound by IgG26AW prevented the further binding of IL-1RI and IL-1RAcP, which confirmed our inference from the result of protein structure. Therefore, the inhibitory mechanism of IgG26AW is to block the assembly of the IL-1β/IL-1RI/IL-1RAcP ternary complex which further inhibits downstream signaling. Based on its high affinity, high neutralizing potency, and novel binding epitope simultaneously occupying both IL-1RI and IL-1RAcP residues that bind to IL-1β, IgG26AW may be a new candidate for treatments of inflammation-related diseases or for complementary treatments of cancers in which the role of IL-1β is critical to pathogenesis.",
keywords = "epitope mapping, human interleukin-1β, IL-1RAcP, IL-1RI, therapeutic antibody",
author = "Kuo, {Wen Chih} and Lee, {Cheng Chung} and Chang, {Ya Wen} and Wei Pang and Chen, {Hong Sen} and Hou, {Shin Chen} and Lo, {Shin Yi} and Yang, {An Suei} and Wang, {Andrew H.J.}",
note = "Funding Information: We are grateful to the National Synchrotron Radiation Research Center (NSRRC), Taiwan, and SPring-8, Japan, for TPS05A and BL44XU beam-time allocations. We thank Dr. Shu-Chuan Jao (Biophysics Core Facility, Scientific Instrument Center, Academia Sinica, Taiwan) and Dr. Meng-Ru Ho (Biophysical Instrumentation Laboratory at the Institute of Biological Chemistry, Academia Sinica, Taiwan) for the BIAcore system analysis and M1000 pro supports. We also thank Dr. Jui-Ling Wang and Dr. Yu-Chia Su in National Laboratory Animal Center (NLAC), NARLabs, Taiwan, for technical supports in animal experimental services. This work was supported by Academia Sinica [AS-SUMMIT-109], the Ministry of Science and Technology [MOST 108-3114-Y-001-002], Taiwan Protein Project [AS-KPQ-105-TPP], and the Technology Supporting Platform Axis [AS-KPQ-106-TSPA]. Funding Information: We are grateful to the National Synchrotron Radiation Research Center (NSRRC), Taiwan, and SPring-8, Japan, for TPS05A and BL44XU beam-time allocations. We thank Dr. Shu-Chuan Jao (Biophysics Core Facility, Scientific Instrument Center, Academia Sinica, Taiwan) and Dr. Meng-Ru Ho (Biophysical Instrumentation Laboratory at the Institute of Biological Chemistry, Academia Sinica, Taiwan) for the BIAcore system analysis and M1000 pro supports. We also thank Dr. Jui-Ling Wang and Dr. Yu-Chia Su in National Laboratory Animal Center (NLAC), NARLabs, Taiwan, for technical supports in animal experimental services. This work was supported by Academia Sinica [AS-SUMMIT-109], the Ministry of Science and Technology [MOST 108-3114-Y-001-002], Taiwan Protein Project [AS-KPQ-105-TPP], and the Technology Supporting Platform Axis [AS-KPQ-106-TSPA]. WC. K and CC. L designed the experiments. WC. K, CC. L, YW. C, W. P, and SY. L executed the experiments. HS. C and SC. H were involved in the antibody screening from phage-display libraries. WC. K and CC. L were involved in the analysis and interpretation of the data and also wrote the manuscript. The authors read and approved the final manuscript. All applicable international, national, and institutional guidelines for the care of animals were followed. All animal protocols were approved by the institutional animal care and use committee of the National Laboratory Animal Center, Taiwan. (IACUC NO: NLAC(TN)-106-D-011R2) (Protocol NO: 106-NLAC-EN-095). This article does not contain any studies with human participants performed by any of authors. The authors declare that they have no competing interests. Publisher Copyright: {\textcopyright} 2020 Elsevier Ltd",
year = "2021",
month = feb,
day = "19",
doi = "10.1016/j.jmb.2020.166766",
language = "English",
volume = "433",
journal = "Journal of Molecular Biology",
issn = "0022-2836",
publisher = "Academic Press",
number = "4",
}
