Development and application of an antibody that binds to interleukin-1β of various mammalian species for the treatment of inflammatory diseases

https://doi.org/10.1016/j.bbrc.2020.04.073Get rights and content

Highlights

  • IL-1β is a main mediator of inflammation among pro-inflammatory cytokines.

  • •7F IgG, an anti IL-1β antibody, binds to IL-1β in human, mouse, rat and canine, specially.

  • 7F IgG alleviates type 2 diabetes and colitis, inflammatory diseases that raises IL-1β.

  • Since 7F IgG can not only bind to IL-1β but also neutralize, it can be used as a therapeutic antibody for the treatment of inflammatory diseases.

Abstract

Inflammation is provoked by host immune reactions to pathogenic or tissue injury and is arbitrated by cytokines. Among the pro-inflammatory cytokines, the tumor necrosis factor α (TNF-α) and interleukin 1β (IL-1β) are main mediators of inflammation. The production of these pro-inflammatory cytokines is mainly triggered in macrophages by harmful stimuli including microbial pathogens, irritants, and toxic cellular components, and plays key roles in the palpation of the inflammatory response. Among the therapeutic antibodies for the treatment of inflammation, those targeting TNF-α (including adalimumab and infliximab) are frequently used in clinical settings. Although IL-1β is a key cytokine for the onset of inflammatory diseases, such as inflammatory bowel disease (IBD) and type 2 diabetes (T2DM), few therapeutic antibodies exist for this cytokine, with the exception of canakinumab. Canakinumab binds to human IL-1β, but does not bind to murine IL-1β, which hampers its experimental use. Therefore, inflammation-therapeutic antibodies that bind to IL-1β of various mammals are needed. In this study, we report the development of an antibody that bound to IL-1β of various mammalian species and exhibited therapeutic effects in inflammatory diseases.

Introduction

Inflammation is becoming increasingly important as it is associated with chronic diseases, such as coronary artery disease, obesity, and cancer, as well as autoimmune and infectious diseases [1]. The role of cytokines in inflammation is well known and has been reported to contribute to autoimmune disorders such as T2DM, IBD, multiple sclerosis and spinal arthrosis. Moreover, anti-cytokine therapies, such as TNF-α neutralizers, are currently common in the treatment of chronic inflammatory diseases [2].

Inflammation is a complex conventional reaction of the body to cell damage and vascularized tissue. The inflammatory reaction, one of the body’s oldest defense mechanism, is controlled by various factors, such as pro-inflammatory cytokines [3]. Pro-inflammatory cytokines are fabricated commonly by triggered macrophages and are associated with the upregulation of inflammatory reactions [4]. Among them, TNF-α and IL-1β are key mediators of inflammation and modulate the development of numerous inflammatory diseases [5].TNF-α, which was identified as a endotoxin-induced glycoprotein in 1975, is produced not only in cells of the monocyte/macrophage lineage, but also in mast cells, T and B lymphocytes, natural killer (NK) cells, neutrophils, endothelial cells, smooth and cardiac muscle cells, fibroblasts, and osteoclasts in inflammatory diseases. The biological effects of TNF-α, among which the regulation of the nuclear factor κB (NF-κB) is key, are distinguished by two distinct receptors, TNFR1 (murine 55 kDa, TNFsfr1a) and TNFR2 (murine 75 kDa, TNFsfr1b). Inflammatory responses associated with tissue damage activated by TNF-α are mainly mediated through TNFR1, whereas TNFR2 mediates tissue repair and angiogenesis, although its signaling is less well characterized than that of TNFR1 [6]. Many antibodies against TNF-α have been developed for the treatment of inflammation, including adalimumab and infliximab, and are used clinically in autoimmune diseases including rheumatoid arthritis and IBD.

The IL-1 cytokine family, which includes IL-1β, is composed of 11 members (seven ligands with agonist activity, three receptor antagonists, and one anti-inflammatory cytokine) and is accepted as a major mediator of inflammation [7]. IL-1β activates macrophages and neutrophils to phagocytose the infesting pathogen and to release toxic oxygen and nitrogen radicals [8]. IL-1β activates the release of the remaining pro-inflammatory cytokines, such as TNF-α and IL-6, and induces Th17 in cell adaptive responses [9]. In vivo, IL-1β is a major cause of acute reactions, including fever and acute protein synthesis [10]. Therefore, IL-1β is a crucial component of the host defense against infections.

As IL-1β is typically activated in situations in which TNF-α is produced [5] and the response rate of anti-TNF-α antibody treatment is low [11], a therapy that targets IL-1β is needed. Canakinumab is an FDA-approved anti IL-1β antibody. Its indications are inflammatory diseases such as systemic juvenile idiopathic arthritis and tumor necrosis factor receptor associated periodic syndrome. Canakinumab binds to human IL-1β but not murine IL-1β, which precludes its testing in various animal models [12]. Therefore, there is a need for antibodies for the treatment of inflammation that can bind to IL-1β of various mammalian species. Here, we present the development of an antibody that bound to IL-1β of various mammalian species and exhibited therapeutic effects in inflammatory diseases.

Section snippets

Materials

We obtained T4 DNA ligase, restriction enzymes, and Vent DNA polymerase, from New England Biolabs (Ipswich, MA, USA). Integrated DNA Technologies (Coralville, IA) supplied the oligonucleotide primers. HRP-conjugated anti-M13 antibody, Dynabeads® M − 270 epoxy, fetal bovine serum and 3,3′,5,5; -tetramethylbenzidine (TMB) single solution were procured from Life Technologies (Carlsbad, CA, USA), GE Healthcare Life-Sciences (Piscataway, NJ, USA), and Thermo Fisher Scientific Inc. (Waltham, MA).

Development of a novel chimeric anti-IL-1β specific monoclonal antibody

We developed a novel chimeric anti-IL-1β specific monoclonal antibody from naïve chicken phage-displayed scFv library. To screen human and mouse IL-1β cross-reactivity clones, a naïve chicken phage-displayed scFv library was biopanned with recombinant human and mouse IL-1β-coated magnetic beads. Rounds 1 and 3 were biopanned with recombinant human IL-1β, and rounds 2 and 4 were biopanned with recombinant mouse IL-1β. After four rounds, the binding reactivities of 96 selected individual clones

Discussion

As IL-1β is a key cytokine in inflammatory diseases, therapies targeting IL-1β are very important. The development of clinically applicable antibodies that neutralize IL-1β relative to TNF-α is rare. Canakinumab, an FDA-approved antibody for treating inflammatory diseases, is difficult to apply to experimental models because it does not bind to murine IL-1β [12]. Therefore, it is necessary to develop an antibody that neutralizes IL-1β and complements the limitations of canakinumab. In addition,

Acknowledgements

This research was supported by Eulji University research grant.

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